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| J Am Acad Dermatol, 1996 Apr, 34(4), 638 - 44 Treatment of photodamaged skin with trichloroacetic acid and topical tretinoin; Humphreys TR et al.; BACKGROUND: Photodamaged skin typically displays lentigines, actinic keratoses, wrinkles, and textural alteration . Chemical peeling has been used to treat these, but few controlled studies have been performed to determine its efficacy . OBJECTIVE: Our purpose was to compare the efficacy of a medium-depth chemical peel with and without tretinoin before and after treatment . METHODS: Sixteen men with actinic damage including actinic keratoses were treated with a 40% trichloroacetic acid(TCA) chemical peel . Half were pretreated for 6 weeks with topical tretinoin; they also used tretinoin after the peel . Photographs were obtained at baseline and at 6 weeks and 6 months after treatment . Changes in specific features were rated by a panel of three examiners . RESULTS: Some improvement was noted in all patients . More rapid and even frosting was observed in the patients pretreated with tretinoin . Solar lentigines, actinic keratoses, and skin texture were the features of photoaging most affected; wrinkles were least affected . No statistically significant difference was found between patients treated with TCA and tretinoin (before and after peel) and those with TCA alone . CONCLUSION: A medium-depth chemical peel with 40% TCA alone produced moderate improvement in some manifestations of actinic damage but had little effect on wrinkles . Treatment with tretinoin before and after TCA did not significantly enhance the efficacy of the peel. Biochim Biophys Acta, 1996 Mar 29, 1300(1), 42 - 8 Identification of an alkaline sphingomyelinase activity in human bile; Nyberg L et al.; The hydrolysis of sphingomyelin has been found to generate important signals regulating cell proliferation, differentiation and apoptosis . However, the enzymes responsible for digestion of dietary sphingomyelin have not been well documented . This study demonstrates the occurrence of a sphingomyelinase (SMase) in both human hepatic bile and gallbladder bile . The enzyme was equally found in both bacteria negative and positive bile samples and in samples obtained from patients with or without gallbladder diseases . A bacteria-free gallbladder bile was used for characterization . It was found that bile SMase hydrolyzed sphingomyelin to phosphorylcholine and ceramide with negligible activity against either phosphatidylcholine or p-nitrophenyl phosphate . The enzyme preferred an alkaline condition and the optimal pH was 9 . The activity of this alkaline SMase was bile salt dependent and was fully activated by 4-6 mM bile salts . Triton X-100, the non-ionic detergent did not activate bile SMase . Ca2+ and Mg2+ ions had no significant effect at optimal bile salt concentration . The molecular mass of this enzyme was about 85 kDa as measured by Sephadex G200 gel chromatography . In conclusion, we demonstrated a SMase in bile which differs markedly from the known acid and neutral SMase . Its potential important roles in sphingomyelin digestion and gallbladder diseases require further investigation. Proc R Soc Lond B Biol Sci, 1996 Mar 22, 263(1368), 257 - 63 Models of the within-host dynamics of persistent mycobacterial infections; Antia R et al.; We use mathematical models to investigate the within-host dynamics of mycobacterial infections . In particular, we investigate the mechanisms by which bacteria such as Mycobacterium tuberculosis and Mycobacterium leprae persist at low densities for extended periods, and attain high densities much later . We suggest that the persistence of bacteria in face of immune pressure may result from the bacteria having a very slow growth rate, or having a dormant stage . We show that whereas these mechanisms may lead to long-term persistence, this will be obtained at relatively low densities . We then suggest that the long-term persistence of bacteria may result in the loss of immunity because of the deletion of specific T-cells arriving from the thymus, and the exhaustion of the specific T-cells as these cells reach the Hayflick limit and die . This loss of immunity will allow the bacteria to attain a high density . We propose experiments capable of testing our models and discuss the implications of the models for the treatment of infected hosts. J Biol Chem, 1996 Mar 22, 271(12), 6998 - 7003 Interaction between the insulin receptor and its downstream effectors . Use of individually expressed receptor domains for structure/function analysis; Paz K et al.; A structural analysis has been carried out to determine which part of the intracellular domain of the insulin receptor (IR) beta subunit is involved in direct interaction with the receptor substrates IRS-1 and Shc . Toward this end, the juxtamembrane (JM) domain (amino acids 943-984) and the carboxyl-terminal (CT) region (amino acids 1245-1 331) of IR were expressed in bacteria as (His)6-fusion peptides, and their interaction with IRS-1 and Shc was studied . We could demonstrate that the CT region of IR was sufficient to bind Shc, although significant, but much lower binding of Shc to the JM region could be detected as well . Furthermore, in vitro Tyr phosphorylation of the CT region potentiated its interactions with Shc 2-fold . In contrast, the JM region, but not the CT domain of the IR, was sufficient to mediate interactions between the IR and IRS-1 . These interactions did not involve the pleckstrin homology (PH) region of IRS-1, since an IRS-1 mutant, in which four "blocks" of the PH domain (Pro5-Pro65) were deleted, interacted with the JM region of IR with the same efficiency as native IRS-1 . These results suggest that the IR interacts with its downstream effectors through distinct receptor regions, and that autophosphorylation of Tyr residues located at the CT domain of the IR can modulate these interactions. Science, 1996 Mar 15, 271(5255), 1519 - 26 Common principles of protein translocation across membranes; Schatz G et al.; Most major systems that transport proteins across a membrane share the following features: an amino-terminal transient signal sequence on the transported protein, a targeting system on the cis side of the membrane, a hetero-oligomeric transmembrane channel that is gated both across and within the plane of the membrane, a peripherally attached protein translocation motor that is powered by the hydrolysis of nucleoside triphosphate, and a protein folding system on the trans side of the membrane . These transport systems are divided into two families: export systems that export proteins out of the cytosol, and import systems that transport proteins into cytosol-like compartments. Nature, 1996 Mar 14, 380(6570), 175 - 9 Selection of RNA-binding peptides in vivo; Harada K et al.; Many priniciples of sequence-specific DNA recognition have been established over the past decade, largely from structural studies of protein-DNA and drug-DNA complexes . On the basis of these principles, it has been possible to design or select variants of known structural motifs, including zinc-fingers and minor groove-binding drugs, that bind desired sequences . Here we describe a strategy, based on transcriptional termination in bacteria, to identify specific RNA-binding peptides using the arginine-rich RNA-binding motif as a framework . Peptides were isolated from two combinatorial libraries that bind tightly and specifically to the Rev response element of HIV . It appears that alpha-helical peptides resembling Rev were selected from one library whereas new peptides that probably do not form helices were selected from the other, suggesting that the arginine-rich motif may be a particularly versatile framework for recognizing RNA structures. Biochemistry, 1996 Mar 12, 35(10), 3175 - 86 Asymmetry requirements in the photosynthetic reaction center of Rhodobacter capsulatus; Taguchi AK et al.; Nine large-scale symmetry reaction center mutants were constructed in Rhodobacter capsulatus by replacing segments of the M subunit gene with the homologous region of the L subunit gene . Between them, the mutations resulted in symmetrization of essentially the entire region from the carboxy terminal portion of the C helix through most of the E helix . The amino acids in this region define about 80% of the environment of the reaction center cofactors . These studies show that roughly 80% of the amino acids that come in close contact with the cofactors involved in initial electron transfer can be made symmetric in a piecewise manner without loss of the ability to grow photoheterotrophically . However, the amino acid regions near the quinones and iron atom are much more sensitive to symmetrization and most of the large-scale changes in this region resulted in the loss of photosynthetic viability, probably due to loss of stable reaction centers from the photosynthetic membrane . More detailed analysis of the isolated photosynthetic membranes from these mutants showed that in all cases but one, there was some amount of charge separation occurring in the mutant reaction centers . This bank of mutants serves as a useful starting point for more detailed studies of the differential molecular interactions which occur between the two reaction center subunits and their associated cofactors. Proc Natl Acad Sci U S A, 1996 Mar 5, 93(5), 2143 - 8 Azotobacter vinelandii NIFL is a flavoprotein that modulates transcriptional activation of nitrogen-fixation genes via a redox-sensitive switch; Hill S et al.; The NIFL regulatory protein controls transcriptional activation of nitrogen fixation (nif) genes in Azotobacter vinelandii by direct interaction with the enhancer binding protein NIFA . Modulation of NIFA activity by NIFL, in vivo occurs in response to external oxygen concentration or the level of fixed nitrogen . Spectral features of purified NIFL and chromatographic analysis indicate that it is a flavoprotein with FAD as the prosthetic group, which undergoes reduction in the presence of sodium dithionite . Under anaerobic conditions, the oxidized form of NIFL inhibits transcriptional activation by NIFA in vitro, and this inhibition is reversed when NIFL is in the reduced form . Hence NIFL is a redox-sensitive regulatory protein and may represent a type of flavoprotein in which electron transfer is not coupled to an obvious catalytic activity . In addition to its ability to act as a redox sensor, the activity of NIFL is also responsive to adenosine nucleotides, particularly ADP . This response overrides the influence of redox status on NIFL and is also observed with refolded NIFL apoprotein, which lacks the flavin moiety . These observations suggest that both energy and redox status are important determinants of nif gene regulation in vivo. Biochemistry, 1996 Mar 5, 35(9), 3093 - 107 Photosystem II of green plants: topology of core pigments and redox cofactors as inferred from electrochromic difference spectra; Mulkidjanian AY et al.; Three electrochromic difference spectra induced by the deposition of (1) a negative charge on the primary quinone acceptor, Q(A), (2) a positive charge on (or near) Tyr161 of the D1 subunit (Y(Z)), and (3) a positive charge on the manganese cluster were determined at room temperature in photosystem II (PSII) core particles from pea . They were deconvoluted into Gaussian components by Powell's numerical optimization procedure . All three spectra were fitted by four components, which we assigned to the Q(y) absorption bands of two chlorophyll a molecules of the primary donor P, the accessory chlorophyll a, and the pheophytin a molecules on the D1 subunit . On the basis of the electrochromic properties of chlorins and our data, we suggest an arrangement of pigments and redox cofactors in PSII that differs from current structural models, which have been shaped like the reaction centers (RC) of purple bacteria . Our model is compatible with sequence data, with the spectroscopic and electrochemical properties of chlorophyll a and pheophytin a, and with the extremely positive redox potential of water oxidation . We conclude the following: (1) P is formed from two orthogonally oriented chlorophyll a molecules that peak at 681 and 677 nm . (2) The accessory chlorophyll a on D1 is oriented perpendicular to the membrane, with ring V pointing to Q(A) . It is presumably attached to His118 of D1 . (3) The mutual arrangement of pheophytin a on the D1 subunit and Q(A) differs from that of their counterparts in bacterial RC . (4) The manganese cluster is located out of the axis that is formed by Y(Z) (Tyr161 of D1), P, and Y(D) (Tyr161 of D2). Biochemistry, 1996 Mar 5, 35(9), 2985 - 91 Flow of structural information between four DNA conformational levels; Levin-Zaidman S et al.; Closed-circular supercoiled DNA molecules have been shown to form a cholesteric assembly within bacteria as well as in vitro under physiological DNA and salt concentrations . Circular dichroism and X-ray scattering studies indicate that the macroscopic structural properties of the chiral mesophase are directly and uniquely dictated by the supercoiling parameters of the constituent molecules . Specifically, we find that the pitch of the DNA cholesteric phase derived from supercoiled DNA is determined by the superhelical density, which, in turn, is modulated by secondary conformational changes . A direct interrelationship among four DNA structural levels, namely, DNA sequence, secondary structural transitions, the tertiary superhelical conformation, and the quaternary, supramolecular organization is accordingly pointed out . Since secondary conformational changes are both sequence and environment dependent, alterations of cellular conditions may effectively modulate the properties of the packed DNA organization, through their effects on secondary structural transitions and hence on the superhelical parameters . On the basis of these results we suggest that liquid crystallinity represents an effectively regulated packaging mode of plectonemic, nucleosome-free DNA molecules in living systems. Schweiz Med Wochenschr, 1996 Mar 2, 126(9), 339 - 47 {Skin manifestations of graft-versus-host reaction following bone marrow transplantation}; Itin PH et al.; We review the cutaneous manifestations of acute and chronic graft versus host disease (GvHD) . Acute GvHD is characterized by initial itching, pain on pressure and erythema which begins on posterior auricular skin, palms and soles . The disease evolves into a typical but nonspecific maculopapular rash . Confluent rashes and follicular erythema may occur . Erosive oral lesions usually develop . The most severe variant of GvHD is toxic epidermal necrolysis, which often has a fatal outcome . The onset of chronic GvHD usually occurs more than 100 days after bone marrow transplantation and may be preceded by the acute form . The spectrum of skin changes includes lichenoid pruritic lesions with violaceous color and scleroderma-like skin involvement . Investigation of unknown rashes in these patients includes skin biopsy, which clearly differentiates leukocytoclastic vasculitis and erythema exsudativum multiforme with lymphocytic vasculitis from cutaneous manifestations of GvHD . Special stains may reveal bacteria and fungus in septicemic patients . The therapeutic options are discussed. Zhonghua Zheng Xing Shao Shang Wai Ke Za Zhi, 1996 Mar, 12(2), 86 - 9 {An experimental study on injury of intestinal immuno-barrier in rat after scald}; Yu Y et al.; A dynamic observation on injury of intestinal immuno-barrier in scalded rat was performed to investigate the relationship between the injury of intestinal immuno-barrier and bacterial translocation . After 40% TBSA third degree scald, a decrease of IgA in intestinal content and the number of CD3+ and CD4+ T lymphocyte, and reduction of IgA coat rate of intestinal bacteria were observed . At the same time, an increase in the incidence of bacterial translocation was detected . The results indicated that intestinal immuno-barrier was damaged and its protective function was weakened after an extensive thermal injury, and it suggested that the injury of intestinal immuno-barrier might play an important role on the development of postburn bacterial translocation and postburn sepsis. Lik Sprava, 1996 Mar-Apr, (3-4), 130 - 4 {The characteristics of the treatment of newly detected pulmonary tuberculosis in miners}; Fomicheva NI et al.; Results are submitted on the treatment of 192 miners free from manifest signs of silicosis when they first presented with different forms of pulmonary tuberculosis . Tuberculosis in those individuals engaged in the coal industry was found out to poorly respond to treatment . During the period of 1.5-2 years, cicatrization of the cavities of decay occurred in 42.6 percent of the cases, no bacteria were recoverable in 50.0 percent . The treatment results were dependent upon the utilization of combinations of antituberculous drug preparations, routes of their administration and choice of pathogenetic options . The greatest benefit from treatment occurred with izoniazid, rifampicin, ethambutol and intrabronchial instillations of izoniazid. Nippon Koshu Eisei Zasshi, 1996 Mar, 43(3), 183 - 95 {A review of epidemiologic studies on the health effects of indoor environments}; Nakai S; Indoor bioaerosols (viruses, bacteria, dust mites, molds, etc.) have become the subject of discussion on indoor environments . A number of epidemiologic studies on the health effects of molds and home dampness, the latter being an important determinant of mold growth, have been conducted mainly in Europe and the United States . In this paper, a review of these epidemiologic studies is described . Questionnaires are used as the method of exposure assessment in most of the studies . The necessity for developing objective methods of dampness/mold exposure assessment has been expressed by some researchers . Respiratory symptoms, especially asthman attacks and wheezing for children, have been utilized in many investigations as a health effect index . Positive relationship between home dampness/mold and respiratory symptoms have been shown in numerous studies . Dampness and/or molds in dwellings may effect respiratory symptoms, but several problems for determining causal inference still remain: exposure assessment, temporal relationship, etc . While the differences in house structure and life style between Japan and Europe/U.S.A . should be considered, the health effects of home dampness and molds need to also be examined in Japan which is a country with high humidity levels. Minerva Stomatol, 1996 Mar, 45(3), 101 - 11 {Saliva and periodontal disease}; Giuliana G et al.; Bacteria and saliva play a very important role in the arising of the oral diseases . It must be considered that the saliva gives the nutriments for the bacteria, and it is able to influence the adhesive mechanisms, and to escape these bacteria . The interaction between the saliva and the host is unknown, but it is well-known in vitro, through the oral ecological systems principles . The authors reviewed several works about the salivary components and their role so in vivo, either in vitro . In this work, first the authors looked for these salivary components, and then, examined the interactions between the saliva, the plaque and the bacteria . So the saliva is a fundamental component in the ecologic succession, and it is nearly related to the etiopathogenesis of periodontitis . Finally a best knowledge of the saliva role and its components in the periodontitis is very interesting, because some salivary components allow the root surfaces colonization, exposed to the oral fluid and they give the possibility to use salivary components as markers of any disease. Rev Sci Tech, 1996 Mar, 15(1), 55 - 71 Husbandry practices employed by private aviculturists, bird markets and zoo collections, which may be conducive to fostering infectious diseases; Wolff PL; Zoos, quarantine stations, and bird markets, dealers and breeders are in the business of propagating or moving birds . Facilities often house and transport birds which have unknown histories of exposure to disease . As few tests are available for disease screening and monitoring in exotic avian species, familiarization with significant avian pathogens will enable the manager and veterinarian to recognize and prevent a disease outbreak . Implementing aviary management practices which minimize the spread of pathogens, in conjunction with quarantine and the proper handling of birds during shipment, can greatly reduce the threat of disease . The author reviews the husbandry practices (caging, nutrition, transport, quarantine) which can reduce the incidence and spread of infectious disease . Significant avian pathogens (bacteria, viruses, parasites) are listed, together with their host range and modes of transmission. Reprod Toxicol, 1996 Mar-Apr, 10(2), 93 - 104 Environmental contaminants in breast milk from the central Asian republics; Lederman SA; Concern has been widespread about possible high levels of environmental contamination in areas of the former Soviet Union . Some of this concern has focussed on toxicants in human milk and their potential adverse effects on the breastfeeding child, but published data have been virtually unavailable . This study collected previously unpublished analyses of breast milk, water, cow's milk, and dairy product samples from several of the Republics during the last decade, providing an opportunity for comparing the level of contamination to similar data from other countries . The general levels of contamination are similar to those observed in other countries . Social and economic conditions as well as the contamination of water and substitute foods in the Republics make alternative methods of infant feeding demonstrably less desirable for the child than the measured levels of breast milk contaminationPIP: Despite widespread concern about high levels of environmental contamination in the former Soviet Union, analyses of breast milk, water, cow's milk, and dairy product samples collected in the past decade from several areas of the Central Asia Republics revealed contamination levels comparable to those in other countries . Of particular concern were industrial and agricultural practices, including the use of pesticides, herbicides, and fertilizers, aimed at increasing productivity . DDT was present in a large portion of breast milk samples in Turkmenistan, Tajikistan, and Kazakhstan . The great range of concentrations of many toxic substances (10- to 20-fold) suggests exposures were from local rather than generalized sources . Seasonal variation in the percentage of positive samples was evident in both breast and cow's milk . Lacking, at present, are population-based studies of breast feeding practices (incidence, daily frequency, duration) in Russia that would enable estimates of the magnitude of infant exposures . For very stable, slowly metabolized, fat-soluble materials such as polychlorinated biphenyls and DDT, breast feeding may contribute disproportionately to an infant's exposure to environmental toxins . It cannot be assumed, however, that exposure will be decreased if infants are bottle fed . Breast feeding avoids exposing infants to bacteria-contaminated water and chemicals in substitute foods . In locales where maternal milk contains environmental toxicants, water and cow's milk are likely to be similarly contaminated . A policy to discourage breast feeding would increase fertility, infant mortality from diarrhea, and household expenditures . Public health professionals concerned about toxic exposure through breast milk should advocate for reduced environmental contamination for people of all ages . J Clin Microbiol, 1996 Mar, 34(3), 734 - 7 Rapid differentiation of Mycobacterium avium and M . paratuberculosis by PCR and restriction enzyme analysis; Eriks IS et al.; Mycobacterium avium subsp . avium (M . avium) and M . avium subsp . paratuberculosis (M . paratuberculosis), intracellular bacteria that can cause chronic granulomatous enteritis in cattle, are difficult to distinguish on the basis of growth and biochemical characteristics . We report the development of a PCR-based strategy for the rapid differentiation of isolates of M . avium from isolates of M . paratuberculosis . Restriction fragment length polymorphism was identified by PCR amplification and subsequent restriction enzyme digestion with PstI of a 960-bp fragment of the 65-kDa heat shock protein (hsp65) from 21 clinical isolates of M . paratuberculosis and 14 isolates of M . avium . These results indicate that a restriction fragment length polymorphism in the hsp65 gene can be used for the rapid differentiation of clinical isolates of M . paratuberculosis and M . avium. J Clin Microbiol, 1996 Mar, 34(3), 543 - 9 Comparison of Bactec 9240 and Difco ESP blood culture systems for detection of organisms from vials whose entry was delayed; Chapin K et al.; A comparison of the Bactec 9240 (Becton-Dickinson, Sparks, Md.) and Difco ESP (Difco Laboratories, Detroit, Mich.) instruments for the detection of organism growth from vials whose entry was delayed was evaluated . The instruments' capabilities for organism recovery, time to detection, rates of false-positive results, and numbers of vials in which growth was not detected were made by using seeded blood culture vial pairs and controls with and without delayed entry . Bactec 9240 and Difco ESP aerobic and anaerobic vials were inoculated with human blood and were seeded with organism growth from 18 species, including obligate aerobic, anaerobic, and facultative anaerobic organisms . Each organism was tested in triplicate at 0, 8, 24, 36, and 48 h and was incubated at both room temperature (RT) and 35 degrees C . Two separate phases of the study were performed, each with a different version of Bactec 9240 software . Overall, detection of growth in vials with delayed entry into either the Bactec 9240 or the Difco ESP instrument resulted in an increased total time to detection with incubation at both RT and 35 degrees C compared with the total time to detection for nondelayed vials . However, false-positive results and vials in which growth was not detected were minimal, and delayed entry did not require routine entry or exit subcultures for either system . Analysis of individual time points and incubation temperatures for the detection of all organisms suggested that Difco ESP vials delayed by up to 8 h may be incubated at 35 degrees C (100% detection) and vials delayed for longer than 8 h may remain at RT . Bactec 9240 vials may be incubated at 35 degrees C for up to 24 h with a minimal loss of detection (97.9% detection), and vials delayed for more than 24 h should remain at RT for optimal recovery of organism growth. Nippon Rinsho, 1996 Mar, 54(3), 685 - 91 {Aquaporin water channels}; Ishibashi K; MIP family proteins with conserved 6 transmembrane domains can be divided into two groups according to their primary sequences . The CHIP group with deletions is predominant in plant and animal kingdom, and functions primarily as water channels . The GLP group without deletions is minor group with limited prevalence and functions as glycerol transporters . Both prototypes are present in bacteria and evolved separately . CHIP group is further divided into three groups: tonoplast aquaporins, plasma membrane aquaporins, and peribacterial nodulins . The recognition of groups in MIP family will facilitate the search for functional domains and deepen our understanding of this ancient protein family. Microbiology, 1996 Mar, 142 ( Pt 3), 675 - 84 A protein having similarity with methylmalonyl-CoA mutase is required for the assimilation of methanol and ethanol by Methylobacterium extorquens AM1; Smith LM et al.; A 4.0 kb region of Methylobacterium extorquens AM1 DNA which complements three mutants unable to convert acetyl-CoA to glyoxylate (and therefore defective in the assimilation of methanol and ethanol) has been isolated and sequenced . It contains two ORFs and the 3'-end of a third one . The mutations in all three mutants mapped within the first ORF, which was designated meaA; it encodes a protein having similarity with methylmalonyl-CoA mutase . However, methylmalonyl-CoA mutase was measured in extracts of one of the mutants and the specific activity was found to be similar to that in extracts of wild-type cells . Furthermore, although the predicted meaA gene product has the proposed cobalamin-binding site, it does not contain a highly conserved sequence (RIARNT) which is present in all known methylmalonyl-CoA mutases; meaA may therefore encode a novel vitamin-B12-dependent enzyme . The predicted polypeptide encoded by the second ORF did not have similarity with any known proteins . The partial ORF encoded a protein with similarity with the 3-oxoacyl-{acyl-carrier-protein} reductases; it was not essential for growth on methanol or ethanol. Microbiology, 1996 Mar, 142 ( Pt 3), 649 - 55 Propionyl-CoA carboxylase from Streptomyces coelicolor A3(2): cloning of the gene encoding the biotin-containing subunit; Bramwell H et al.; In Streptomyces coelicolor A3(2), polyketides are made from malonyl-CoA, which is presumed to be derived from acetyl-CoA by the action of acetyl-CoA carboxylase (ACC) . No ACC activity was found in cell-free extracts of S . coelicolor . However, propionyl-CoA carboxylase (PCC) activity was detected at substantial levels . Fixation of CO2 by ACC and PCC occurs by covalent bonding of CO2 to a biotin-containing protein . Most bacteria have a single small biotinylated protein of approximately 22 kDa, but S . coelicolor contains three larger biotin-containing proteins (approximately 145, 88 and 70 kDa) . To determine which biotinylated protein was associated with PCC activity, the enzyme was purified and shown to comprise an alpha subunit (biotin-containing) of 88 kDa and a beta subunit of 66 kDa . The N-terminal sequences of these proteins were determined and, using an oligonucleotide probe, the gene for the alpha subunit (pccA) was cloned. Sarcoidosis Vasc Diffuse Lung Dis, 1996 Mar, 13(1), 38 - 42 Pulmonary sarcoidosis: could mycoplasma-like organisms be a cause? Johnson LA, Edsall JR, Austin JH, Ellis K. The etiology of sarcoidosis is unknown but an unusual bacterial agent is possible . Mycoplasma-Like Organisms {MLO} are obligate intracellular cell wall deficient bacteria with a distinctive ultrastructural appearance . MLO are a common cause of various transmissible plant diseases . Despite over 25 years of effort MLO remain uncultivated . Molecular biologic studies indicate MLO are only distantly related to extracellular cultivable mycoplasma . Diagnosis of MLO diseases is based chiefly on detection of the organisms in infected cells by electron microscopy . Recently MLO have been detected by electron microscopy within leucocytes in sterile inflamed aqueous and vitreous humor from patients with idiopathic chronic uveitis including sarcoidosis uveitis . Preliminary molecular biologic studies suggest that human MLO are quite closely related phylogenetically to plant MLO . Inoculation of human uveitis MLO into mouse eyelids produced chronic uveitis and lethal systemic granulomatous disease with MLO within leucocytes and endothelial cells in the disease sites . The MLO induced animal pulmonary disease resembled sarcoidosis . This report describes abnormal intracellular bodies consistent with MLO within leucocytes and endothelial cells adjacent to the granulomas in transbronchial biopsies from 9 corticosteroid untreated sarcoidosis patients versus none in 4 control lungs. Br Poult Sci, 1996 Mar, 37(1), 73 - 85 Mortality in near-term ostrich embryos during artificial incubation; Brown CR et al.; 1 . The ostrich industry in South Africa (and elsewhere) experiences a high rate of embryo mortality during artificial incubation of eggs . Most of this mortality takes place in the last l0-l4 d of incubation . 2 . We carried out post-mortem examinations on 111 embryos that died within this period to assess the causes of this mortality . 3 . Malpositioning and severe oedema were the predominant symptoms of dead-in shell embryos with 55% being malpositioned and 41% showing severe oedema . Of these, 22 embryos (24%) showed both symptoms . Malpositioning generally results from incorrect setting of the eggs or inadequate turning and oedema was significantly correlated with the amount of water lost from the eggs which in turn was correlated with egg size . 4 . Myopathy, gross lesions of internal organs, haemorrhage, bacterial infections and congenital deformities were found in less than 10% of chicks examined for these symptoms. Immunol Today, 1996 Mar, 17(3), 131 - 7 A new foreign policy: MHC class I molecules monitor the outside world; Rock KL; Although most cells exclusively use their major histocompatibility complex (MHC) class I molecules to present peptides from endogenous proteins, phagocytes also use them to present exogenous antigens . Here, Kenneth Rock describes how this novel antigen-presenting pathway may play an important role in immune surveillance for intracellular bacteria or parasites, as well as for viral infections and tumors affecting somatic tissues. Protein Expr Purif, 1996 Mar, 7(2), 183 - 93 Purification of a secreted form of recombinant rabies virus glycoprotein: comparison of two affinity tags; Wojczyk BS et al.; Expression of recombinant eukaryotic proteins in transfected mammalian cell lines has become an important approach for the characterization of the structure and function of these proteins . However, it is often difficult to recover and purify the recombinant proteins . Therefore, the use of fusion proteins incorporating epitope or affinity tags has become more widespread . In this paper, we directly compare two affinity tags, the hexahistidyl tag and the biotin peptide mimetic, Strep-tag, for use in purification of a recombinant soluble form of rabies virus glycoprotein secreted by transfected Chinese hamster ovary fibroblasts . The recombinant rabies virus glycoproteins are denoted RGP(WT)T441his and RGP(WT)T443s-tag, respectively . These affinity tags were chosen because the chromatographic matrices (Ni(II)-NTA-agarose and recombinant core streptavidin-agarose, respectively) were readily available and these methods offered the possibility of a one-step purification using mild elution conditions . However, in our hands, neither method allowed for a one-step purification protocol . Nonetheless, it was possible to purify RGP(WT)T441his to homogeneity from crude conditioned medium using a combination of metal-chelate affinity chromatography and immunoaffinity chromatography . In contrast, although the Strep-tag has been useful for purifying recombinant proteins expressed in bacteria, we were not able to effectively purify RGP(WT)T443s-tag from conditioned medium using chromatography on recombinant core streptavidin-agarose. J Dermatol Sci, 1996 Mar, 11(3), 214 - 22 Secretory component (polymeric immunoglobulin receptor) expression on human keratinocytes by stimulation with interferon-gamma and differences in response; Nihei Y et al.; Secretory IgA (sIgA) is a major protective factor in the mucosal immune system because of its great ability to form complexes with bacteria . Secretory component (SC) is an 80-kDa glycoprotein, a component of sIgA, which functions as a polymeric immunoglobulin receptor for IgA and aids the secretion of sIgA from the epithelial surface . We studied SC production by keratinocytes which were involved in the inflammatory process using interferon-gamma (IFN-gamma) as one of the major inflammatory promoters produced by helper T cells . Using two human squamous cell carcinoma cell lines (HSCs) and normal human keratinocytes (NHKs), results from flow cytometric analysis, enzyme-linked immunosorbent assay (ELISA), and Northern blotting revealed that HSCs produced SC when stimulated with IFN-gamma, although their responses differed; one line exhibited enhanced SC production whereas the production in the other line was suppressed . NHKs also exhibited SC expression on the cell surface by means of immunocytochemical analysis, flow cytometry and ELISA, however the responses were also different in each strain . Although the reason for the diversity of SC expression on keratinocytes is not clear, these differences may influence epidermal sIgA secretion level. J Physiol Pharmacol, 1996 Mar, 47(1), 79 - 90 13C-urea breath test for detection of Helicobacter pylori and its correlation with endoscopic and histologic findings; Hilker E et al.; The urea breath test (UBT) reflects the intragastric urease activity and thus the extent of H . pylori (H.p.) burden of the mucosa . We compared UBT results with gastroscopic and histologic findings in 174 patients . H.p.density, grade and activity of the chronic gastritis type B were semiquantitatively assessed . 46% of patients were H.p.-positive . A type B gastritis was significantly (p < 0.01) associated with H.p . colonization . UBT-results significantly correlated with the grade (r = 0.53) and the activity (r = 0.45) of gastritis . They correlated better with grade and activity of the inflammation than did the H.p.-density in histology (r = 0.38 and r = 0.37) . There is a considerable interindividual variability in UBT-results for the same degree of gastritis indicating that besides the number of bacteria other factors must be of significance for the severity of inflammation . Endoscopic findings apart from ulcers do not predict the presence of H.p. J Physiol Pharmacol, 1996 Mar, 47(1), 195 - 202 The stimulation and inhibition of T cell proliferation by Helicobacter pylori components; Chmiela M et al.; The purified T cells from peripheral blood of healthy human volunteers, seronegative for anti-Helicobacter pylori antibody were stimulated in cultures with live or heat-killed H . pylori rods or with bacterial sialic acid-specific surface haemagglutinin (sHA), a crude surface (SF) or cytoplasmic (CF) fractions . It is demonstrated that H . pylori bacteria contain both stimulatory and inhibitory components for T cells of healthy individuals . The sHA as well as SF (5-20 micrograms) induced the proliferative response of T lymphocytes . By contrast, CF inhibited in dose dependent manner, the proliferation of T cells in the cultures stimulated with H . pylori bacteria or PHA . The result suggest that in vivo, a dominance of activation or immunosuppression could depend on the concentration of the bacteria and their products in infective foci. Am J Pathol, 1996 Mar, 148(3), 825 - 35 Characterization of a panel of novel anti-p21Waf1/Cip1 monoclonal antibodies and immunochemical analysis of p21Waf1/Cip1 expression in normal human tissues; Fredersdorf S et al.; As a universal inhibitor of cyclin-dependent kinases and one of the target genes of the tumor suppresser p53, p21Waf1/Cip1 can act as a tumor suppresser through its ability to control cell cycle progression . To study the function of p21Waf1/Cip1 protein and to investigate its tissue distribution, a panel of anti-p21Waf1/Cip1 monoclonal antibodies was generated . These anti-p21Waf1/Cip1 monoclonal antibodies were initially raised against a GST-p21Waf1/Cip1 fusion protein produced in bacteria . Detailed characterization of the antibodies showed that they can specifically detect p21Waf1/Cip1 by immunoblotting, immunoprecipitation, and immunostaining . The specific induction of p21Waf1/Cip1 expression in response to gamma-radiation in cells containing p53 was also detected by these antibodies . The ability to detect p21Waf1/Cip1 expression in conventionally fixed tissue sections allowed us to investigate the distribution of p21Waf1/Cip1 in 23 different types of normal human tissues, and p21Waf1/Cip1 expression was found in most tissues . A close inverse relationship between p21Waf1/Cip1 expression and proliferation was seen in some tissues, including gastrointestinal tract . However, such association is not universal . In tissues such as lung, kidney, thyroid, pancreatic ducts and acini, and liver, despite the fact that most of the cells are quiescent, expression of p21Waf1/Cip1 was detected only in occasional epithelial cells . All these suggest that the expression of p21Waf1/Cip1 varies among different human tissues . Finally, epitope mapping of the anti-p21Waf1/Cip1 antibodies using a peptide library covering the entire p21Waf1/Cip1 protein sequence indicates that two of the antibodies recognize a region of p21Waf1/Cip1 close to that bound by proliferating cell nuclear antigen . These two monoclonal antibodies will therefore be additionally useful in further understanding the functions of p21Waf1/Cip1 both in vitro and in vivo. In Vivo, 1996 Mar-Apr, 10(2), 249 - 53 Crowding; Burger H et al.; Crowding is the situation in which case a large number of animals, including man, are restricted in environmental space . Such an occurrence may start with just a few animals . In plants, including bacteria in culture, agriculture may be considered as an artificial example of this process . We see the ontogenic forms of crowding developing from stress into superorganisms (such as certain social insects) and from stress into pathological crowding in animals, plants and man . Crowding is shown by environmental changes such as in the introduction of new species of certain rodents and lagomorphs into the Australian environment . Aspects of crowding play a role during the housing of zoos, domestic or laboratory animals . Space limitations and overpopulation initiates stress and infections such as tuberculosis and tumours . This is found in man himself in the development of slums through to urbanisation as seen in Asia particularly, in prisons, in the workplace, in the school or home. Genet Anal, 1996 Mar, 12(5-6), 197 - 200 Using electroporation and a slot cuvette to deliver plasmid DNA to insect embryos; Leopold RA et al.; Microinjection is the method used almost exclusively to deliver DNA constructs to insect embryos while electroporation is commonly used for DNA delivery to bacteria, cell cultures and certain plant tissues . This communication describes a method using an easily constructed slot cuvette and the electroporation technique for transfer of DNA to insect embryos for possible use in developing methods for germline transformation . This method eliminates time-consuming individual embryo manipulation and thus far has been found to be adaptable for use on several types of insect embryos . Using this method, we show successful transfer of plasmid DNA to embryos of the corn earworm moth, Helicoverpa zea, and the house fly, Musca domestica. Electrophoresis, 1996 Mar, 17(3), 512 - 7 Quantitation of polymerase chain reaction-amplified DNA fragments by capillary electrophoresis and laser-induced fluorescence detection; Vincent U et al.; Quantitation of DNA fragments amplified by polymerase chain reaction (PCR) is needed for the determination of target DNA in molecular biology . Capillary electrophoresis in entangled polymer solution coupled to laser-induced fluorescence detection was assessed as an alternative technique to conventional slab gel methods to monitor competitive PCR, which consists of amplifying an internal standard fragment under the same conditions as the target fragment . The fluorescence signal was generated either through end-labeling of the fragments using 5'-fluorescein-labeled primers or through intercalation of ethidium bromide along the double strand . It is shown that the more accurate and reliable results were obtained using this latter pathway. Pediatr Dent, 1996 Mar-Apr, 18(2), 139 - 44 Evaluation of an alum-containing mouthrinse in children for plaque and gingivitis inhibition during 4 weeks of supervised use; Putt MS et al.; Aluminum salts have demonstrated activity against oral bacteria and also have shown indications of inhibiting plaque and gingivitis . The aims of this clinical trial were to determine the effects of daily supervised rinsing with a specially formulated, alum-containing mouthrinse on existing plaque and gingivitis in children and to monitor its effect on the oral tissues and its acceptability to subjects . Using a double-blind, parallel design, 48 sixth-graders rinsed once daily for 30 sec under supervision with either a placebo or a mouthrinse containing 0.02 M alum while continuing their normal oral hygiene habits . Plaque and gingivitis were assessed and intraoral examinations were performed at 0, 2 and 4 weeks . The alum mouthrinse significantly (P < 0.05) reduced the amount of plaque relative to the placebo after both 2 and 4 weeks . Gingivitis and plaque thickness also were decreased, but the differences did not attain significance . No evidence of deleterious effects to the oral tissues was observed and the alum mouthrinse was well accepted by the children . This trial demonstrated that daily use of an alum-containing mouthrinse was safe and produced a significant effect on plaque that supplemented the benefits of daily toothbrushing . Thus, topically applied aluminum may have potential applications in preventive dentistry for controlling plaque. J Clin Periodontol, 1996 Mar, 23(3 Pt 1), 160 - 4 Effect of localized experimental gingivitis on early supragingival plaque accumulation; Daly CG et al.; Previous investigations have reported increased plaque formation in the presence of gingival inflammation as compared with gingival health . In these investigations, experimental gingivitis has been induced by total abolition of oral hygiene such that increased levels of salivary bacteria could contribute to the increased plaque formation . In the present study, experimental gingivitis was localized to 2 selected teeth whilst maintaining normal oral hygiene measures in the rest of the mouth . The aim of this study was to investigate whether the mass of plaque accumulating in a 3-day period is influenced by the inflammatory status of the adjacent gingival margins . Following scaling, prophylaxis and a period of optimal hygiene to establish gingival health, "plaque-guards" were worn during routine oral hygiene performance to prevent any cleaning on the buccal surfaces of teeth 14 and 15 from days 0 to 3 . On day 3, accumulated plaque was removed from a 1.5 mm zone on the buccal surfaces of 14 and 15 adjacent to the gingival margins and weighed immediately . Subjects continued to wear their plaque-guards during oral hygiene until day 14 to induce experimental gingivitis around the experimental teeth . At day 14, all plaque was cleaned from the buccal surfaces of the experimental teeth and the mass of plaque accumulating over the next 3 days weighed on day 17 . Comparison of plaque weights showed that the wet-weight of 3 day-old plaque was higher in the presence of experimental gingivitis than in the presence of gingival health (p = 0.02) . This observation suggests that the inflammatory status of the marginal gingiva has an important effect on early, supragingival plaque accumulation. J Anim Sci, 1996 Mar, 74(3), 663 - 71 Spent cellulose casings as potential feed ingredients for ruminants; Gentry JL et al.; Cellulose casings are used to contain and form meat and poultry emulsions during the smoking and cooking process . Casings then are stripped from the cooked product and traditionally disposed of in landfills . Because of the bulk of the spent cellulose casings (SCC), rapid composting technology may be used to reduce bulkiness . The following SCC were evaluated in vitro and in vivo: fibrous ground (FG), fibrous composted (FC), NOJAX ground (NG), and NOJAX composted (NC) . In vitro digestibility was determined by incubating SCC with mixed ruminal bacteria for 0, 6, 12, 24, 36, 48, and 72 h . In vivo data were collected using four ruminally cannulated Holstein steers in a 4 x 4 Latin square design . Diets consisted of a 50:50 ratio of alfalfa hay-wheat middlings with 5% cornsteep liquor . Diets contained no SCC (CON) or 25% (DM basis) of the FC, FG, or NC SCC substrate . Casings were high in structural carbohydrate and salt content but low in CP, ether extract, and lignin concentrations . In vitro OM digestibility at 24 h was highest (P < .05) for FC and lowest (P < .05) for NG; FG and NC were intermediate . Composting tended to reduce fiber content and increase digestion . In vivo intakes and digestibilities were not adversely affected by inclusion of SCC in the diet . Thus, SCC have the ability to partially replace more traditional forages, such as alfalfa hay and wheat middlings, in high-fiber diets for growing beef cattle . Limitations in the use of SCC as a partial replacement of traditional feedstuffs will likely be because of high salt concentrations in the casings resulting from product brine chilling. Zh Mikrobiol Epidemiol Immunobiol, 1996 Mar-Apr, (2), 10 - 3 {The capacity of avirulent forms of Francisella tularensis for dissemination and proliferation in the host body}; Tsimbalistova MV et al.; In this investigation isogenic avirulent variants obtained from F . tularensis standard virulent strain 503 were used . The capsule-deficient variants (cap-) were shown to have no species-specific capsular antigens and to be capable of producing R-LPS having no the polysaccharide part of the molecules . The capsule-defective forms (cap +/- ) were found to synthesize capsular antigens and S-LPS whose polysaccharide part essentially differed from the O-lateral chains of LPS of the virulent strain . The study of bacterial dissemination revealed that virulent bacteria rapidly spread in the macroorganism, and their subsequent proliferation shortly led to the death of animals . Avirulent mutants (cap- and cap +/- ) appeared in the organs of animals later and proliferated slower, parasitizing in the macroorganism without fatal outcome . The cap- variants were not capable of inducing the synthesis of antitularemic antibodies and possessed no protective properties . The cap +/- mutants were capable of inducing the synthesis of antitularemic antibodies in mice . These antibodies facilitated the elimination of avirulent strains from the macroorganism, but did not ensure protection from infection with virulent strains. Infect Dis Clin North Am, 1996 Mar, 10(1), 93 - 9 Fever in the elderly; Norman DC et al.; Fever in elderly persons is only one clinical presentation that can be used to assist the clinician at suspecting a serious disease, such as an infection . Infections, like all other illnesses in the geriatric patient, may occur with a variety of nonspecific, atypical, nonclassic, and unusual manifestations . The clinician caring for elderly patients should be aware of these nonclassical presentations of infections in this age group . Unexplained change in functional capacity, worsening of mental status, weight loss or failure to thrive, weakness and fatigue, falls, and generalized pain are only some of the clues that may aid the clinician in considering infection in elderly persons . Key concepts of fever in older adults are: Fever generally indicates presence of serious infection, most often caused by bacteria . Fever may be absent in 20%-30% of elderly patients harboring a serious infection . Criteria for fever in elderly patients should also include an elevation of body temperature of at least 2 degrees F from baseline values . FUO in elderly persons is caused by infections (30%-35%), CTD (25%-30%), and malignancies (15%-20%) in the majority of cases. J R Soc Med, 1996 Mar, 89(3), 170P - 1P Munchausen's syndrome--masquerading as necrotizing fasciitis; Park AJ et al.; We would like to present a novel form of Munchausen's syndrome which presented us with a difficult problem in the midst of the recent media hysteria regarding flesh eating bacteria . This condition, first described by Richard Asher in 1951, is often very difficult to diagnose and it is not, therefore, surprising that the victims have been through several episodes of treatment before the suspicions of staff are raised . It owes its name to a fictitious character, Baron Karl Friedrich Hieronymus Freihess von Munchausen, who recounted extraordinary and wildly exaggerated exploits of adventure and daring, not unlike patients who present with this condition . Our enquiries revealed that the laid down procedure when faced with a patient with this condition is that the regional medical officer (RMO) should be contacted . It is then his responsibility to contact other RMOs, who in turn pass the information on to accident and emergency consultants in their area. J Mol Evol, 1996 Mar, 42(3), 359 - 68 Relationships between transposable elements based upon the integrase-transposase domains: is there a common ancestor? Capy P, Vitalis R, Langin T, Higuet D, Bazin C. The integrase domain of RNA-mediated elements (class I) and the transposase domain of DNA-mediated transposable elements (class II) were compared . A number of elements contain the DDE signature, which plays an important role in their integration . The possible relationships between mariner-Tc1 and IS elements, retrotransposons, and retroviruses were analyzed from an alignment of this region . The mariner-Tc1 superfamily, and LTR retrotransposons and retroviruses were found to be monophyletic groups . However, the IS elements of bacteria were found in several groups . These results were used to propose an evolutionary history that suggests a common ancestor for some integrases and transposases. J Surg Res, 1996 Mar, 61(2), 521 - 6 Effect of endotoxemia on intestinal villus microcirculation in rats; Schmidt H et al.; Intestinal mucosal hypoperfusion with subsequent ischemia during endotoxemia might cause a breakdown of the gut barrier with translocation of bacteria and their toxins into the systemic circulation, thus maintaining a "gut-derived" septic state . The aim of this study was to investigate the influence of endotoxin on the microcirculation of intestinal villi, which represent the most vulnerable part of the mucosa . The changes in blood flow and in the diameters of the central villus arterioles located in the distal ileum were monitored in control rats without lipopolysaccharide (LPS) exposure (n=7), and in rats receiving 1.5 mg/kg b.w . LPS (n=7) or 15 mg/kg b.w . LPS (n=7) over 60 min . The blood flow and the arteriolar diameters were determined using in vivo videomicroscopy at baseline, and 60 min and 120 min later . In control animals, no change in blood flow and arteriolar diameters were observed during the entire experiment . Administration of 1.5 mg/kg b.w . LPS reduced the blood flow to 69.5 +/- 9.0% of the baseline value at the end of the study period . This decrease in blood flow was associated with a decrease in the villus arteriolar diameters by 17.4 +/- 2.5% from the baseline values . In animals exposed to 15 mg/kg b.w . LPS, the decrease in villus blood flow at 60 min was 64.8 +/- 10.9% of baseline, and at 120 min 66.9 +/- 12.6% of baseline . The diameters of the villus arterioles were reduced by 11.5 +/- 2.4% and 15.1 +/- 1.7%, respectively . In the control group and in the 1.5-mg/kg LPS group, the mean arterial blood pressure did not change during the entire study period . In the 15-mg/kg LPS group, the mean arterial pressure tended to decrease after 60 min . These data suggest a reduction of villus blood flow due to vasoconstriction in the central villus arterioles during normotensive endotoxmia, which might represent the mechanism for the mucosal ischemia observed in critically ill patients. J Surg Res, 1996 Mar, 61(2), 433 - 6 Infectious sequelae in the use of polyglycolic acid mesh for splenic salvage with intraperitoneal contamination; Wolf SE et al.; Salvage of the injured spleen is important in the trauma patient . Loss of the spleen can result in both early and late infectious complications due to immunologic and phagocytic deficits . Splenic salvage techniques include the use of polyglycolic acid (PGA) mesh to wrap and tamponade the damaged and bleeding spleen . However, the use of mesh may increase the incidence of infection in the presence of intraperitoneal contamination . We examined whether mesh in the contaminated field increases the infection rate compared to splenectomy in a murine model . Sixty male Sprague-Dawley rats were divided into three groups of 20 each: splenectomy, splenic wrap with PGA, and control (with splenic mobilization) . All rats were subjected to a standard inoculum of enteric bacteria at the time of celiotomy . Sixteen (80%) of the splenectomy rats, 10 (50%) of the PGA mesh wrapped rats, and four (20%) of the control rats expired (P < 0.5) . In surviving rats, necropsy at 7 days demonstrated abscess formation in all four (100%) of splenectomy, four of 10 (40%) in PGA mesh wrapped, and two of 16 (13%) of control rats . All of the abscesses in the wrap group involved the mesh . Overall infection rates (including fatal peritonitis, abscess formation, and empyema) were 100% for splenectomy, 75% for PGA mesh wrapped, and 30% for control rats (P < 0.05) . We conclude in this experimental model that the use of PGA mesh wrap does increase susceptibility to infection, but much less so than splenectomy in the presence of intraperitoneal contamination. Int J Dermatol, 1996 Mar, 35(3), 181 - 4 Clinical-epidemiologic study of alopecia areata; Roselino AM et al.; BACKGROUND . Alopecia areata is a common disease and may be associated with autoimmune disease, atopy, Down syndrome, emotional stress, and foci of sepsis . METHODS . Seven cases of alopecia areata were diagnosed among workers in the Water and Effluent Treatment Sector (WETS) of a paper factory, representing a 0.6% incidence, when the value for the population at large is 0.1% . Three of these workers are assigned to the WETS on a permanent basis and four provide maintenance services . One of the latter patients had alopecia areata that fully regressed . Because biologic treatment of water and effluents involves saprophytic bacteria and fungi as well as chemical substances such as acrylamide, a clinical examination and laboratory tests were performed on all workers assigned permanently to the WETS (N = 9) and on 25% of the workers, selected at random providing services to the sector (N = 14) . RESULTS . There was no association between alopecia areata and atopy, dermatophytosis, or bacteria isolated . Toxicologic evaluation revealed an acrylamide-like substance in 7 workers with alopecia areata, with a statistically significant correlation . Measures were taken at the workplace to decrease worker contact with the mists (probably containing acrylamide) in the pulp-pressing room; no other cases of alopecia areata had been detected 1 year after the study . CONCLUSIONS . A survey of the literature did not show reports of alopecia areata as an occupational dermatosis, but our conclusion is, that this dermatosis could be due to the professional activities of the workers at the paper factory studied. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 1996 Mar, 81(3), 333 - 42 Diffuse sclerosing osteomyelitis and florid osseous dysplasia; Groot RH et al.; The literature on diffuse sclerosing osteomyelitis of the mandible has included at least two groups of lesions: (1) those from which bacterial infectious agents are rarely isolated (chronic-tendoperiostitis); and (2) those from which bacteria are readily isolated (true diffuse sclerosing osteomyelitis) . The latter should be distinguished from secondarily infected florid osseous dysplasia . In this article the features of 16 patients with sclerotic jawbone lesions associated with symptoms of infection are analyzed . Eleven patients showed a large area of sclerosis of the mandible that was not restricted to the alveolar process and was surrounding an infectious focus . The histologic pattern revealed a deposition of reactive bone . These lesions are considered to represent true diffuse sclerosing osteomyelitis . The remaining five patients showed sclerotic lesions restricted to the alveolar process in one or more quadrants of the jaws . Apart from inflammation and reactive changes, histologic pattern revealed a fibroblastic stroma with bone and cementum-like structures that are formed by metaplasia . These lesions are considered to represent secondarily infected florid osseous dysplasia. FASEB J, 1996 Mar, 10(4), 444 - 52 Nicotinamide nucleotide transhydrogenase: a model for utilization of substrate binding energy for proton translocation; Hatefi Y et al.; The energy-transducing nicotinamide nucleotide transhydrogenases of mammalian mitochondria and bacteria are structurally related membrane-bound enzymes that catalyze the direct transfer of a hydride ion between NAD(H) and NADP(H) in a reaction that is coupled to transmembrane proton translocation . The protonmotive force alters the affinity of the transhydrogenase for substrates, accelerates the rate of hydride ion transfer from NADH to NADP, and shifts the equilibrium of this reaction toward NADPH formation . Transhydrogenation in the reverse direction from NADPH to NAD is accompanied by outward proton translocation and formation of a protonmotive force . In reverse transhydrogenation, the enzyme utilizes substrate binding energy for proton pumping . Therefore, with regard to the mechanism of energy transduction, the transhydrogenase works according to the same principles as the ATP synthase complex of mitochondria and bacteria, the proton and cation ATPases, and possibly certain redox-linked proton pumps . However, the relatively simple structure of the transhydrogenase recommends it as a model for study of the utilization of binding energy for vectorial translocation of protons and other cations. Infect Immun, 1996 Mar, 64(3), 1030 - 4 Antibody responses to Brucella abortus 2308 in cattle vaccinated with B . abortus RB51; Stevens MG et al.; Cattle vaccinated with Brucella abortus rough strain RB51 (SRB51) produced small amounts of serum immunoglobulin G (IgG) but no IgM antibody to smooth strain 2308 (S2308) bacteria and produced no IgG or IgM antibody to S2308 lipopolysaccharide (LPS) . Western immunoblot analysis revealed that antiserum from SRB51-vaccinated cattle contained IgG antibody that reacted with S2308 proteins of 84 to <20 kDa . However, antiserum from the vaccinated cattle did not contain agglutinating B . abortus antibody in the tube agglutination test for brucellosis . These results suggest that SRB51-vaccinated cattle produced no antibody to S2308 LPS, although they did produce nonagglutinating IgG antibody that reacted with S2308 bacteria and bacterial proteins of 84 to <20 kDa. Burns, 1996 Mar, 22(2), 101 - 6 Early effects of smoke inhalation on alveolar macrophage functions; Bidani A et al.; Alveolar macrophage (AM) dysfunctions have been implicated in the pathogenesis of smoke inhalation lung injury . We investigated the early (within 70 min) effects of smoke inhalation on AM . The cells were recovered by bronchoalveolar lavage from rabbits ventilated with cotton smoke for 5 min followed by O2/room air for 60 min (smoke-exposed) or with room air in place of smoke (control) . Smoke injury caused arterial blood carboxyhaemoglobin levels to increase 11-fold and reduced arterial blood PO2 (measured approximately 1 h postinjury) by 25 per cent . Scanning electron micrographs revealed denudation of plasmalemmal pseudopods in smoke-exposed AM . Smoke exposure suppressed both AM adherence to plastic and phagocytosis of opsonized bacteria . Basal superoxide (O2-) production was elevated in smoke-exposed AM, compared with controls, whereas PMA-stimulated O2- production was unaffected . Smoke-exposed AM had reduced basal secretion of tumour necrosis factor-alpha (TNF-alpha), but displayed a greater TNF response to stimulation with LPS than did control cells . LPS-stimulated TNF-alpha releases from control and smoke-exposed AM were suppressed by phosphodiesterase inhibitors pentoxifylline and theophylline, and were enhanced by the lipoxygenase inhibitor, MK886 . The early responses of AM to smoke inhalation lung injury are consistent with activation of O2- production and priming of TNF-alpha release, concurrent with a functional down regulation of phagocytosis. J Bacteriol, 1996 Mar, 178(5), 1469 - 72 Carboxylation of epoxides to beta-keto acids in cell extracts of Xanthobacter strain Py2; Allen JR et al.; A novel enzymatic reaction involved in the metabolism of aliphatic epoxides by Xanthobacter strain Py2 is described . Cell extracts catalyzed the CO2-dependent carboxylation of propylene oxide (epoxypropane) to form acetoacetate and beta-hydroxybutyrate . The time courses of acetoacetate and beta-hydroxybutyrate formaton indicate that acetoacetate is the primary product of propylene oxide carboxylation and that beta-hydroxybutyrate is a secondary product formed by the reduction of acetoacetate . Analogous C5 carboxylation products were identified with 1,2-epoxybutane as the substrate . In the absence of CO2, propylene oxide and 1,2-epoxybutane were isomerized to form acetone and methyl ethyl ketone, respectively, as dead-end products . The carboxylation of short-chain epoxides to beta-keto acids is proposed to serve as the physiological reaction for the metabolism of aliphatic epoxides in Xanthobacter strain Py2. Nord Med, 1996 Mar, 111(3), 74 - 6, 83 {Diagnosis and treatment of conjunctivitis}; Ehlers N et al.; Conjunctivitis is a frequent reason for consulting a doctor . Most often treatment consists of antibiotics, even though sensitive bacteria are only rarely demonstrated . In our part of the world conjunctivitis is self-limiting, and it may be worth considering the effect of treatment, if any . With this in mind, diagnosis and differential diagnoses are reviewed and with regard to treatment distinctions are made between neonatal conjunctivitis (gonococcus, chlamydia, virus), conjunctivitis in children (symptoms of upper respiratory tract infections), and conjunctivitis in adults and the elderly (dry eyes, epiphora in ectropion). Naturwissenschaften, 1996 Mar, 83(3), 103 - 12 {Evolution of cells}; Maier UG et al.; Life has existed on earth for some 4 x 10(9) years . During most of this time, evolution took place at the level of cell evolution . The cells of presently existing organisms belong to two fundamentally different cell types, protocytes (of bacteria and archaea) and eucytes (of eukarya) . Thanks to molecular phylogenetics, the path of evolution can now be traced back to its very beginnings, although the picture may be blurred by repeated horizontal gene transfer . A symbiogenetic origin of plastids and mitochondria is now very well documented, and it is being discussed also for some other constituents of eucytes, including even the cells nucleus . It could be demonstrated that not only did bacterial cells become incorporated into protoeucytes and transformed into organelles of their respective hosts, but also that endocytic eucytes have apparently been transformed to complex organelles by coevolution with host cells. J Biol Chem, 1996 Mar 1, 271(9), 4850 - 7 Function of multiple heme c moieties in intramolecular electron transport and ubiquinone reduction in the quinohemoprotein alcohol dehydrogenase-cytochrome c complex of Gluconobacter suboxydans; Matsushita K et al.; Alcohol dehydrogenase (ADH) of acetic acid bacteria functions as the primary dehydrogenase of the ethanol oxidase respiratory chain, where it donates electrons to ubiquinone . ADH is a membrane-bound quinohemoprotein-cytochrome c complex which consists of subunits I (78 kDa), II (48 kDa), and III (14 kDa) and contains several hemes c as well as pyrroloquinoline quinone as prosthetic groups . To understand the role of the heme c moieties in the intramolecular electron transport and the ubiquinone reduction, the ADH complex of Gluconobacter suboxydans was separated into a subunit I/III complex and subunit II, then reconstituted into the complex . The subunit I/III complex, probably subunit I, contained 1 mol each of pyrroloquinoline quinone and heme c and exhibited significant ferricyanide reductase, but no Q1 reductase activities . Subunit II was a triheme cytochrome c and had no enzyme activity, but it enabled the subunit I/III complex to reproduce the Q1 and ferricyanide reductase activities . Hybrid ADH consisting of the subunit I/III complex of G . suboxydans ADH and subunit II of Acetobacter aceti ADH was constructed and it had showed a significant Q1 reductase activity, indicating that subunit II has a ubiquinone-binding site . Inactive ADH from G . suboxydans exhibiting only 10% of the Q1 and ferricyanide reductase activities of the active enzyme has been isolated separately from active ADH (Matsushita, K., Yakushi, T., Takaki, Y., Toyama, H., and Adachi, O (1995) J . Bacteriol . 177, 6552-6559) . Using these active and inactive ADHs and also isolated subunit I/III complex, we performed kinetic studies which suggested that ADH contains four ferricyanide-reacting sites, one of which was detected in subunit I and the others in subunit II . One of the three ferricyanide-reacting sites in subunit II was defective in inactive ADH . The ferricyanide-reacting site remained inactive even after alkali treatment of inactive ADH and also after reconstituting the ADH complex from the subunits, in contrast to the restoration of Q1 reductase activity and the other ferricyanide reductase activities . Thus, the data suggested that the heme c in subunit I and two of the three heme c moieties in subunit II are involved in the intramolecular electron transport of ADH into ubiquinone, where one of the two heme c sites may work at, or close to, the ubiquinone-reacting site and another between that and the heme c site in subunit I . The remaining heme c moiety in subunit II may have a function other than the electron transfer from ethanol to ubiquinone in ADH. Ann Otol Rhinol Laryngol, 1996 Mar, 105(3), 234 - 41 Experimental otitis media with Moraxella (Branhamella) catarrhalis; Fulghum RS et al.; Two hundred fifty gerbils and 7 chinchillas were utilized in 11 experiments to determine the effect of inoculating viable and heat-killed suspensions of Moraxella (Branhamella) catarrhalis into the middle ear cavity . Development of otitis media was observed by otoscopy and histopathology . Gerbils were found to be susceptible to 2 x 10(4) viable M catarrhalis cells . Depending on the number of cells inoculated, the resulting untreated inflammation was a very mild to moderately severe, self-limiting disease with no permanent sequelae except in animals inoculated with high {10(6) to 10(7)} numbers of bacteria . Viable bacteria could not be isolated from the middle ears 24 hours after inflammation was induced . Heat-killed cells produced less severe acute inflammation with no permanent sequelae . We conclude 1) gerbils and chinchillas are susceptible to a self-limited inflammation caused by M catarrhalis, 2) no infection occurs, since viable bacteria cannot be recovered from middle ear aspirates, and 3) viable cells produce more severe inflammation than heat-killed cells. EMBO J, 1996 Mar 1, 15(5), 1163 - 71 Instability of long inverted repeats within mouse transgenes; Collick A et al.; Various sequences in the mammalian genomes are unstable . One class of sequence arrangement is long inverted repeats, which are known to be unstable in bacteria and yeast . While in mammals some evidence suggests that short inverted repeats (<10 bp long) may show instability, nothing is known about the stability of long inverted repeats . Here we describe two unrelated multicopy transgenes in the mouse (loci 109 and OX1-5), each of which contains a long inverted repeat that shows substantial mitotic instability . This instability also occurs in the germline so that mutant transgenes appear within pedigrees at a high frequency . The mutation processes acting at these two inverted repeats are complex and can involve insertion or deletion, and can result in stabilization of the transgene . At transgene 109 mutational events range from very small rearrangements at the centre of the inverted repeat to complete transgene deletion . In addition we show that the rates of mutation at the inverted repeat of transgene OX1-5 can vary between the male and female germlines and between inbred strains of mice, suggesting the possibility of a genetic analysis to identify loci that modulate inverted repeat instability. J Leukoc Biol, 1996 Mar, 59(3), 357 - 62 Macrophage dysfunction following the phagocytosis of IgG-coated erythrocytes: production of lipid peroxidation products; Loegering DJ et al.; The phagocytosis of erythrocytes may contribute to the increased susceptibility to life-threatening infections in patients with burn injury, sickle cell anemia, and malaria . The phagocytosis of immunoglobulin G-coated erythrocytes (EIgG) is followed by a transient depression of several macrophage functions including phagocytosis, respiratory burst capacity, and killing of bacteria . The present study suggests the possibility that after erythrophagocytosis hemoglobin-derived iron conspires with reactive oxygen products of the macrophage respiratory burst to cause oxidant damage to the phagocyte . Challenge of elicited peritoneal macrophages with EIgG phagocytosis was followed by an increase in lipid peroxidation as assessed by thiobarbituric acid-reactive substances (TBARS) . Doses of EIgG associated with increased TBARS also caused a depression of Fc receptor-mediated phagocytosis and phorbol myristate acetate (PMA)-stimulated hydrogen peroxide production . Time course experiments demonstrated that the increase in TBARS coincided with the depression of macrophage function . There was no increase in TBARS following the phagocytosis of IgG-coated erythrocyte ghosts, suggesting that hemoglobin iron is involved in the generation of TBARS . The phagocytosis of erythrocyte ghosts did not depress macrophage function . Since complement receptor-mediated phagocytosis does not stimulate the respiratory burst, the role of the respiratory burst in causing lipid peroxidation was assessed using the phagocytosis of complement-coated erythrocytes . Phagocytic challenge with complement-coated erythrocytes caused neither an increase in TBARS nor a depression of macrophage function . However, there was an increase in TBARS when the respiratory burst was stimulated with PMA following complement receptor-mediated phagocytosis of erythrocytes . These results suggest that hemoglobin iron and phagocyte-generated oxidants collaborate to cause the depression of macrophage function following EIgG phagocytosis. Arch Microbiol, 1996 Mar, 165(3), 201 - 5 Purification and characterization of chlorobenzene cis-dihydrodiol dehydrogenase from Xanthobacter flavus 14p1; Spiess E et al.; Chlorobenzene cis-dihydrodiol dehydrogenase was purified to homogeneity from Xanthobacter flavus 14p1, which used 1,4-dichlorobenzene as the sole source of carbon and energy . The enzyme converted a number of halogenated substrates with high specific activity . The pI of the native chlorobenzene cis-dihydrodiol dehydrogenase was 5.4, and the molecular mass was approximately 100 kDa, as determined by gel filtration . The enzyme was composed of four apparently identical subunits with a molecular mass of 26.5 kDa . The Michaelis constant for 3,6-dichlorobenzene cis-dihydrodiol (210 microM) was lower than for benzene cis-dihydrodiol (780 microM), while the specific activity with benzene cis-dihydrodiol (63 units/ mg) was higher than with 3,6-dichlorobenzene cis-dihydrodiol (32 units/mg) . Chlorobenzene cis-dihydrodiol dehydrogenase accepted also NADP+ as cosubstrate; however, the activity was reduced to 14% of that with NAD+ . The enzymic activity was inhibited by mercuric chloride and to a lesser extent by the metal-ion chelators 8-hydroxyquinoline and KCN. J Mol Biol, 1996 Feb 16, 256(1), 31 - 49 Two competing pathways for self-splicing by group II introns: a quantitative analysis of in vitro reaction rates and products; Daniels DL et al.; Self-splicing group II introns are found in bacteria and in the organellar genes in plants, fungi, and yeast . The mechanism for the first step of splicing is generally believed to involve attack of a specific intronic 2'-hydroxyl group on a phosphodiester linkage at the 5'-splice site, resulting in the formation of a lariat intron species . In this paper, we present kinetic and enzymatic evidence that in vitro there are two distinct pathways for group II intron self-splicing: one involves 2'-OH attack and another involves attack of water or hydroxide . These two pathways occur in parallel under all reaction conditions, although either can dominate in the presence of particular salts or protein cofactors . Both pathways are followed by a successful second step of splicing, and either pathway can be highly efficient . We find that the hydrolytic pathway prevails under physiological ionic conditions, while branching predominates at molar concentrations of ammonium ion . The intron is observed to adopt two major active conformations . In order to quantify their individual reaction rates, we applied a mechanistic model describing biphasic parallel kinetic behavior . Kinetic analysis throughout the investigation reveals that there is no coupling between the unproductive "spliced-exon-reopening" reaction (SER) and hydrolysis during the first step of splicing . Conditions that stimulate branching can promote the SER reaction just as efficiently as conditions that stimulate the hydrolytic pathway . Although there is little evidence that it exists in vivo, a hydrolytic splicing pathway for group II introns has important implications for the translation of intron-encoded proteins and the inhibition of intron migration into new genomic positions. FEMS Microbiol Lett, 1996 Feb 15, 136(2), 209 - 13 Immunochemical characterization of a haemagglutinating antigen of Arcobacter spp; Tsang RS et al.; The Arcobacter haemagglutinin has been identified by Western immunoblot to be an immunogenic protein of about 20 kDa . The haemagglutinating activity is sensitive to proteolytic enzyme digestion and heat treatment of 80 degrees C and above . The Arcobacter haemagglutinin is possibly a lectin-like molecule binding to erythrocytes via a glycan receptor containing D-galactose as part of its structure. Oncogene, 1996 Feb 15, 12(4), 727 - 34 Reduced phosphotyrosine binding by the v-Src SH2 domain is compatible with wild-type transformation; Tian M et al.; The SH2 domain of v-Src binds phosphotyrosyl-proteins in vivo and in vitro . The function of this domain is necessary for transformation of Rat-2 cells and for morphologically wild-type transformation of chicken embryo fibroblasts (CEF) . The phosphate group of phosphotyrosine interacts directly with a conserved arginine residue in the FLVRES motif of the SH2 domain, R175 in v-Src . To examine the role of phosphotyrosine binding in transformation by v-Src, we have characterized the effects of R175 mutations on the transforming ability of v-Src and on the interaction of the v-Src SH2 domain with phosphotyrosyl-proteins . The R175H mutation, and to a lesser extent the R175K mutation, reduced but did not eliminate the binding of phosphotyrosyl-proteins to the v-Src SH2 domain . However neither mutation affected transformation of CEF or Rat-2 cells by v-Src and neither mutation resulted in major changes in the level or pattern of protein-tyrosine phosphorylation in transformed CEF . In contrast, the R175E mutant of v-Src induced fusiform transformation of CEF and failed to transform Rat-2 cells; the mutant SH2 domain was insoluble when expressed in bacteria, suggesting that the R175E mutation disrupts the structure of the v-Src SH2 domain . We conclude that, although the Arg residue in the FLVRES motif is invariant in most if not all SH2 domains, at position 175 in the v-Src SH2 domain residues other than arginine can support the binding of phosphotyrosyl-proteins, albeit at reduced levels . Furthermore under the expression conditions normally used, that is when v-Src is expressed under the control of a retroviral LTR, the reduced binding of phosphotyrosyl-proteins is compatible with wild-type transformation. Eur J Biochem, 1996 Feb 15, 236(1), 171 - 9 Identification of a functional cAMP response element in the secretogranin II gene; Cibelli G et al.; Secretogranin II is an acidic secretory protein with a widespread distribution in secretory granules of neuronal and endocrine cells . The secretogranin II gene contains, like other members of the granin family, a cAMP response element (CRE) in its upstream region . To investigate the functional significance of this motif, intracellular cAMP levels were increased in a neuronal cell line derived from the septal region of the brain and the level of secretogranin II gene expression was analysed . It was found that increased cAMP levels did, in fact, induce secretogranin II gene expression . To analyse the cis-acting sequence responsible for this induction, a hybrid gene containing the upstream region of the mouse secretogranin II gene fused to beta-globin as a reporter was constructed . Transfection analysis revealed that cAMP-induced transcription of the secretogranin II promoter/beta-globin gene in septal and insulinoma cells . DNA-protein binding assays showed that recombinant CRE-binding protein (CREB), produced in bacteria or human cells, bound in a sequence-specific manner to the secretogranin II promoter CRE . Moreover, deletion mutagenesis revealed that the CRE motif is a bifunctional genetic regulatory element in that it mediates basal as well as cAMP-stimulated transcription . Interestingly, cAMP had no effect upon secretogranin II gene transcription in PC12 and neuroblastoma cells . An increase in the intracellular cAMP concentration activated a GAL4-CREB fusion protein upon transcription in neuroblastoma cells indicating the integrity of the cAMP signaling pathway to the nucleus . Basal as well as cAMP-stimulated transcription, directed from the secretogranin II promoter was, however, impaired in insulinoma cells by overexpression of CREB-2, a negative-acting CRE-binding protein . These results indicate that competitive effects are likely to occur between CRE-bound transcriptional activators and repressors . We conclude that cAMP-stimulated induction of secretogranin II gene transcription is mediated by the CRE motif in a cell-type-specific manner, and is likely to depend on the balance between positive and negative CRE-binding proteins in a particular cell type. Eur J Biochem, 1996 Feb 15, 236(1), 144 - 8 Crystallization and crystal packing of recombinant 3 (or 17) beta-hydroxysteroid dehydrogenase from Comamonas testosteroni ATTC 11996; Benach J et al.; The enzyme 3 (or 17) beta-hydroxysteroid dehydrogenase from Comamonas testosteroni was crystallized . Crystals, of up to 0.6 mm in their longest dimension and suitable for a crystallographic analysis have been obtained by the vapour diffusion method . They belong to the orthorhombic lattice type and diffract to a maximum resolution of 0.23 nm . A final data set obtained by merging data from three crystals resulted in a completeness of 90% with an Rmerge of 6% . A molecular replacement search carried out by using 3 alpha (or 20 beta)-hydroxysteroid dehydrogenase from Streptomyces hydrogenans as a search model allowed us to assign I222 as the correct space group and to propose a model for the crystal packing, with one monomer per asymmetric unit . Thus, the whole unit cell contains two tetramers . The R-factor after rigid body refinement is 48.1%. J Clin Invest, 1996 Feb 15, 97(4), 915 - 24 Interleukin-11: stimulation in vivo and in vitro by respiratory viruses and induction of airways hyperresponsiveness; Einarsson O et al.; To address the role of IL-11 in viral airways dysfunction, we determined whether infectious agents that exacerbate asthma stimulate stromal cell IL-11 production, determined whether IL-11 could be detected at sites of viral infection and evaluated the effects of IL-11 on airway physiology . Respiratory syncytial virus (RSV), parainfluenza virus type 3 (PIV3), and rhinovirus (RV) 14 were potent stimulators while cytomegalovirus and adenovirus only weakly stimulated and herpes simplex virus type 2 and bacteria did not stimulate IL-11 elaboration . IL-11 was not detected or barely detected in nasal aspirates from children without, but was detected in aspirates from children with viral upper respiratory tract infections . The levels of IL-11 were highest in patients with clinically detectable wheezing . IL-11 also caused nonspecific airways hyperresponsiveness in BALB/c mice . These studies demonstrate that three major causes of viral-induced asthma, RSV, RV, and PIV, in contrast to other viruses and bacteria, share the ability to induce stromal cell IL-11 production . They also demonstrate that IL-11 can be detected in vivo during viral respiratory infections, that the presence of IL-11 correlates with clinical bronchospasm and that IL-11 is a potent inducer of airways hyperresponsiveness . IL-11 may be an important mediator in viral airways disorders. FEBS Lett, 1996 Feb 12, 380(1-2), 142 - 6 Role of the mitochondrial DnaJ homologue, Mdj1p, in the prevention of heat-induced protein aggregation; Prip-Buus C et al.; The role of the mitochondrial Hsp70 system in the prevention of heat-induced protein aggregation was studied in isolated mitochondria from Saccharomyces cerevisiae . Firefly luciferase was employed as a thermolabile tester protein . After shift to 40 degrees Celsius transient increase of mt-Hsp70/luciferase complex was observed, which required functional Mdj1p and Mge1p, the mitochondrial homologues of DnaJ and GrpE . The kinetics of luciferase aggregation, however, were not influenced by mutations in either mt-Hsp70 or Mge1p . Only the absence of Mdj1p led to enhanced protein aggregation . Thus, a central role in the transient protection against heat stress is attributed to this mitochondrial DnaJ homologue. J Biol Chem, 1996 Feb 9, 271(6), 2909 - 13 Induction, localization, and purification of a novel sialidase, deaminoneuraminidase (KDNase), from Sphingobacterium multivorum; Nishino S et al.; Recently, we reported the discovery of a new type of sialidase, KDNase, which specifically hydrolyzes the ketosidic linkages of 2-keto-3-deoxy-D-glycero-D-galacto-nononic acid (KDN), but not N-acylneuraminyl linkages . We now report that this enzyme, designated KDNase SM, is an inducible enzyme that is localized in the periplasm of Sphingobacterium multivorum . Growth of S . multivorum in the presence of KDN-containing oligosaccharide alditols, KDNalpha2-->3Galbeta1-->3GalNAc alpha1-->3{KDNalpha2--> (8KDN alpha2-->)n-->6}GalNAcol, as a sole carbon source induced KDNase SM activity 15 40-fold, compared with growth in the absence of inducer . KDN, Neu5Ac, or Neu5Ac oligomers were ineffective as inducers . The enzyme was released from the periplasm of induced cells by cold osmotic shock and purified 700-fold to homogeneity . The specific activity of the pure enzyme was 82,100 units/mg of protein . KDNase SM activity resided in a single polypeptide chain with an estimated molecular weight of approximately 47,500 . Enzyme activity was maximal at near neutral pH . The availability of pure KDNase will now make it possible to study the structure and functional role of KDN-glycoconjugates and to determine the molecular mechanism whereby the enzyme can discriminate between KDN and N-acylneuraminic acid. Mol Divers, 1996 Feb, 1(2), 87 - 96 Structural aspects of antibody-antigen interaction revealed through small random peptide libraries; Slootstra JW et al.; Two small random peptide libraries, one composed of 4550 dodecapeptides and one of 8000 tripeptides, were synthesized in newly developed credit-card format miniPEPSCAN cards (miniPEPSCAN libraries) . Each peptide was synthesized in a discrete well (455 peptides/card) . The two miniPEPSCAN libraries were screened with three different monoclonal antibodies (Mabs) . Two other random peptide libraries, expressed on the wall of bacteria (recombinant libraries) and composed of 10(7) hexa- and octapeptides, were screened with the same three Mabs . The aim of this study was to compare the amino acid sequence of peptides selected from small and large pools of random peptides and, in this way, investigate the potential of small random peptide libraries . The screening of the two miniPEPSCAN libraries resulted in the identification of a surprisingly large number of antibody-binding peptides, while the screening of the large recombinant libraries, using the same Mabs, resulted in the identification of only a small number of peptides . The large number of peptides derived from the small random peptide libraries allowed the determination of consensus sequences . These consensus sequences could be related to small linear and nonlinear parts of the respective epitopes . The small number of peptides derived from the large random peptide libraries could only be related to linear epitopes that were previously mapped using small libraries of overlapping peptides covering the antigenic protein . Thus, with respect to the cost and speed of identifying peptides that resemble linear and nonlinear parts of epitopes, small diversity libraries based on synthetic peptides appear to be superior to large diversity libraries based on expression systems. Quintessence Int, 1996 Feb, 27(2), 129 - 35 Scanning electron microscopic evaluation of resin-dentin and calcium hydroxide-dentin interface with resin composite restorations; Goracci G et al.; Calcium hydroxide has been used as a liner in resin composite restorations to protect the pulp . Recent research has demonstrated that pulpal inflammation is caused by microleakage of restorations and by the subsequent passage of bacteria . The present study involved scanning electron microscopic observation of cross-sections of resin composite-dentin interfaces after the interposition of a layer of calcium hydroxide . A new-generation adhesive system that involves etching of the dentin was used . Ultrastructural analysis indicated that polymerization shrinkage of the resin composite caused the separation of the calcium hydroxide from the dentinal surface, forming 8- to 15-micron-wide interfacial gaps in 100% of the areas studied . Where the adhesive was applied directly to dentin, it adhered closely, forming a gap-free attachment with evidence of an acid-resistant hybrid layer (4 to 6 microns in thickness) and resin tags of various lengths that hermetically sealed the dentinal tubules. An Med Interna, 1996 Feb, 13(2), 59 - 63 {Role of bronchoalveolar lavage and transbronchial biopsy in the diagnosis of pneumonia in patients with organ transplantation}; Arnedillo Munoz A et al.; Pneumonia in patients with organ transplantation constitutes a very frequent cause of mortality, as a result precocious aetiologic diagnosis is indispensable . The bronchoscopic techniques, bronchoalveolar lavage (BAL) and transbronchial biopsy (TBB), constitute fundamental procedures for these diagnoses . We begin this study with the aim of evaluating the profitability obtained with these procedures . 36 bronchoscopies were performed on 29 patients with organ transplantation, in all of them we realized BAL and in 20 TBB . We confirm the presence of pneumonia in 30 (in 15 of them we had performed TBB), the BAL was diagnostic in 20 cases (66.6%) and the TBB in 7 (46.6%) . With both, BAL and TBB, we obtained a sensitivity of 80% and a specificity of 75% . We isolated 10 bacteria, 8 Citomegalovirus (CMV), 6 Pneumocystis carinii and 2 Aspergillus fumigatus . The BAL and the TBB contributed significantly in the aetiologic diagnosis of pneumonia in patients with organ transplantation, consequently we consider them basic tools in the management of these infections. Lett Appl Microbiol, 1996 Feb, 22(2), 141 - 4 Use of monoclonal antibodies against dibenzo-p-dioxin degrading Sphingomonas sp . strain RW1; Thakur IS; A monoclonal antibody prepared against surface antigen of Sphingomonas sp . strain RW1 was used for the direct detection of RW1-like organisms in environmental samples by epifluorescence microscopy and subsequent confirmation by Western blot . Of the 76 samples collected from various sources and probed using epifluorescence, only one sample, effluent from paper and pulp processing, gave a positive result . The effluent was cultured and yielded an organism which, by Western blot analysis, was shown to contain the 28 kDa protein recognized by the monoclonal antibody. Immunol Cell Biol, 1996 Feb, 74(1), 81 - 9 Manipulation of the helper T cell response to influence antigenic competition occurring with a multivalent vaccine; Hunt JD et al.; The reduction in antibody observed following inoculation with multiple heterologous Dichelobacter nodosus pili antigens is thought to be due to competition between antigen-specific B cells for a limited amount of T cell help . We demonstrate here that this competition is not further influenced by the expansion of cross-reactive antibody secreting cells at the expense of serogroup specific antibody secreting cells . The T cell determinants of pili recognized by sheep and BALB/c mice have been defined using 15 residue peptides . These T cell determinants include cross-reactive determinants in the conserved amino terminal region of the antigen . Here we investigate the effect of expanding the pili-specific T cell population by priming with pili derived T cell determinants . It was not possible to increase the antibody elicited in response to the multivalent vaccine by priming mice with either a synthetic peptide spanning a T cell determinant or with reduced and alkylated or heterologous serogroups of pili 4 weeks before inoculation with the multivalent vaccine . A strategy designed to increase the T cell population by inoculating animals with pili covalently coupled to an extrinsic T cell determinant was pursued. Immunol Cell Biol, 1996 Feb, 74(1), 8 - 25 ISCOMs (immunostimulating complexes): the first decade; Barr IG et al.; A little over a decade ago, novel immunostimulating complexes (ISCOMs) were described . This review examines the position and progress that ISCOM technology has achieved in the fields of vaccine research and medicine over this period . Much of the work on ISCOMs has remained in the area of vaccine research where there is still an urgent need for improved adjuvants to help combat important diseases such as AIDS, malaria and influenza . Currently the only widely licensed adjuvants for human use are the aluminium salts, but with the trend towards highly purified subunit vaccines, which are inherently less immunogenic than some of the older vaccines, potent adjuvants capable of promoting specific immune responses are required . ISCOMs are one such technology that offers many of these requirements and as their use in vaccines enters its second decade clinical trials are commencing that will establish whether these submicron, non-living particles composed of saponin, cholesterol, phospholipid and in many cases protein, are useful components for a range of human vaccines. Mol Biochem Parasitol, 1996 Feb-Mar, 76(1-2), 125 - 34 Characterization of iron-dependent endogenous superoxide dismutase of Plasmodium falciparum; Becuwe P et al.; Two main superoxide dismutase activities at isoelectric points (pI) 6.2 and 6.8 and two minor at pI 5.6 and 6.4 were found in crude extracts of Plasmodium falciparum . These activities were cyanide-resistant and hydrogen peroxide-sensitive and represented 20-30% of the total SOD activity found in the crude extract . A fragment of 424 bp, amplified from genomic DNA from P . falciparum, was cloned and sequenced . The deduced amino acid sequence identified this fragment as a coding region of an SOD gene . A cDNA corresponding to SOD was then isolated from a P . falciparum cDNA library and sequenced . The deduced amino acid sequence of SOD (197 aa) was compared with 32 known Feor Mn-SODs by the 'DARWIN' system . This analysis showed that the parasitic enzyme was related to typical Fe-SODs . The SOD subunit was purified and the N-terminal sequence, determined up to 29 residues, corresponded to that of cDNA isolated . The iron-dependent SOD activity found in Plasmodium falciparum represents the first level of the antioxidant defence system of the parasite . It is also the first SOD characterized in the parasitic Apicomplexa phylum whose sequence can be compared to equivalent iron-dependent enzymes known in other protozoa and bacteria. J Mol Evol, 1996 Feb, 42(2), 194 - 200 Phylogenetic relationships of nonaxenic filamentous cyanobacterial strains based on 16S rRNA sequence analysis; Nelissen B et al.; In order to determine the nearly complete 16S rRNA gene sequences of cyanobacteria originating from nonaxenic cultures, a cyanobacterium-specific oligonucleotide probe was developed to distinguish polymerase chain reaction (PCR) products of the cyanobacterial rRNA operons from those resulting from amplification of contaminating bacteria . Using this screening method the 16S rRNA genes of four nonaxenic filamentous cyanobacterial strains belonging to the genera Leptolyngbya and Oscillatoria were cloned and sequenced . For the genus Leptolyngbya, the 16S rRNA sequence of the axenic strain PCC 73110 was also determined . Phylogenetic trees were constructed based on complete and partial sequences . The results show that the strains Leptolyngbya foveolarum Komarek 1964/112, Leptolyngbya sp . VRUC 135 Albertano 1985/1, and Leptolyngbya boryanum PCC 73110 belong to the same cluster . Strain Oscillatoria cf . corallinae SAG 8.92, which contains the rare photosynthetic pigment CU-phycoerythrin, is not closely related to other CU-phycoerythrin-containing cyanobacteria . Oscillatoria agardhii CYA 18, which is a representative of planktonic Oscillatoria species that form toxic blooms in Norwegian inland waters, has no close relatives in the tree. FEMS Microbiol Lett, 1996 Feb 1, 136(1), 79 - 84 Gene sequence analysis and properties of EGC, a family E (9) endoglucanase from Fibrobacter succinogenes BL2; Bera C et al.; The endoglucanase gene (endC) of Fibrobacter succinogenes BL2 encodes a protein of 620 amino acids (EGC) that shows similarity with family E1 cellulases, and particularly with EGB from F . succinogenes S85 . Alignment of the amino acid sequence of family E1 cellulases revealed that EGC is composed of a N-terminal domain and a large catalytic domain of 453 residues containing an extension of 60 residues at its C-terminal part which is not present in other family E1 enzymes . EGC shows the same substrate specificity as EGB, and is also inhibited by EDTA . However, its optimal pH (7.0) and temperature (37 degrees C) for activity are different. Biol Chem Hoppe Seyler, 1996 Feb, 377(2), 71 - 86 Cystatins in health and disease; Henskens YM et al.; Proteolytic enzymes have many physiological functions in plants, bacteria, viruses, protozoa and mammals . They play a role in processes such as food digestion, complement activation or blood coagulation . The action of proteolytic enzymes is biologically controlled by proteinase inhibitors and increasing attention is being paid to the physiological significance of these natural inhibitors in pathological processes . The reason for this growing interest is that uncontrolled proteolysis can lead to irreversible damage e.g . in chronic inflammation or tumor metastasis . This review focusses on the possible role of the cystatins, natural and specific inhibitors of the cysteine proteinases, in pathological processes. Appl Microbiol Biotechnol, 1996 Feb, 44(6), 801 - 6 Anaerobic dechlorination and mineralization of pentachlorophenol and 2,4,6-trichlorophenol by methanogenic pentachlorophenol-degrading granules; Kennes C et al.; Anaerobic granules developed for the treatment of pentachlorophenol (PCP) completely mineralized 14C-labeled PCP to 14CH4 and 14CO2 . Release of chloride ions from PCP was performed by live cells in the granules under anaerobic conditions . No chloride ions were released under aerobic conditions or by autoclaved cells . Addition of sulfate enhanced the initial chloride release rate and accelerated the process of mineralization of 14C-labeled PCP . Addition of molybdate (10 mM) inhibited the chloride release rate and severely inhibited PCP mineralization . This suggests involvement of sulfate-reducing bacteria in PCP dechlorination and mineralization . Addition of 2-bromoethane sulfonate slightly decreased the chloride release rate and completely stopped production of 14CH4 and 14CO2 from {14C}PCP . 2,4,6-trichlorophenol was observed as an intermediate during PCP dechlorination . On the basis of experimental results, dechlorination of 2,4,6-trichlorophanol by the granules was conducted through 2,4-dichlorophenol, 4-chlorophenol or 2-chlorophenol to phenol at pH 7.0-7.2. Semin Immunol, 1996 Feb, 8(1), 3 - 9 Anti-red blood cell autoantibody transgenic mice: murine model of autoimmune hemolytic anemia; Murakami M et al.; We established an anti-red blood cell (RBC) autoantibody transgenic mouse line, in which almost all B cells were deleted in the periphery . A small number of B-1 cells, however, escaped from deletion, survived and expanded in the peritoneal cavity and the gut, because of the absence of RBC . The activation of B-1 cells by enteric bacteria induced autoimmune hemolytic anemia (AIHA) . In turn, AIHA was cured by elimination of peritoneal B-1 cells . This Tg mouse line is useful for revealing the generation and activation of B-1 cells, and for clarifying the physiological and pathological roles of B-1 cells. Antimicrob Agents Chemother, 1996 Feb, 40(2), 488 - 90 Enzymatic method for inactivation of aminoglycosides during measurement of postantibiotic effect; den Hollander JG et al.; To determine the postantibiotic effect of aminoglycosides, two methods are currently being used to remove the test drug: repeated washing and dilution . An enzymatic inactivation method of removing gentamicin and tobramycin was developed and compared with the dilution method . This enzymatic method provides a rapid and simple alternative method of removing aminoglycosides which results in reliable postantibiotic-effect values. Biochem Mol Med, 1996 Feb, 57(1), 1 - 9 Batten disease and mitochondrial pathways of proteolysis; Tanner AJ et al.; Most forms of Batten Disease (BD), a group of neurodegenerative diseases, are characterized by the accumulation within lysosomes of the very hydrophobic protein subunit 9 of the mitochondrial F1F0-ATP synthase (F-ATPase) . It is now known that the cause of the accumulation of this protein in BD is a reduction in its rate of degradation . Because the F-ATPase subunit 9 accumulates within lysosomes of BD tissues, the degradative defect seemed likely to be within lysosomes . However, a recent report showed that delayed degradation of F-ATPase subunit 9 was evident in fibroblasts from BD patients long before any of the protein could be found within lysosomes . Therefore, the defective degradation pathway in BD appears likely to be intramitochondrial . We review the rather limited information about pathways of degradation of mitochondrial proteins . Mitochondria can be taken up and degraded by lysosomes through a process called macroautophagy . However, substantial proteolysis also occurs within mitochondria . Several different proteases are present within mitochondria, but their normal protein substrates are largely unknown . Like proteases from bacteria, many of these proteases operate in concert with molecular chaperones . We hypothesize that a mutation in a gene encoding a mitochondrial protease or a mitochondrial molecular chaperone leads to impaired degradation of F-ATPase subunit 9 in BD . This proteolipid may then form intracellular aggregates that are eventually sequestered into lysosomes. Jpn J Med Sci Biol, 1996 Feb, 49(1), 15 - 27 Effective isolation of MPB64 from a large volume of culture filtrate of Mycobacterium bovis BCG Tokyo; Haga S et al.; MPB64, a secretory protein of Mycobacterium bovis BCG Tokyo, was isolated from a culture filtrate of the bacteria in Sauton synthetic medium harvested on day 8 . The protein was isolated by five steps; (i) concentration of the culture filtrate by cutting the molecules smaller than 5 kDa with the Millipore Pellicon Cassette system, (ii) affinity separation by a Phenyl Sepharose CL-4B column, (iii) separation with a DEAE-Sepharose CL-6B column with 3 M urea, (iv) separation with a Sephacryl S200HR column, and (v) separation with a DEAE-Sepharose column without urea . MPB64 in each fraction was determined by comparing the band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with that of standard MPB64 . The specificity of isolated MPB64 was tested by immunoblotting with anti-MPB64 antibody . The potency of isolated MPB64 in eliciting skin reaction in the BCG-sensitized guinea pigs was the same to that of standard MPB64 . The method described herein is an improved one for isolating MPB64 from a large volume of culture filtrate of M . bovis BCG Tokyo . The technique should be applicable to isolation of other mycobacterial secretory proteins. Curr Opin Cell Biol, 1996 Feb, 8(1), 30 - 7 The actin-related proteins; Frankel S et al.; A family of proteins has been discovered over the past three years whose members have clear sequence homology to actin but are distinguished from actin by their structural and functional diversity . The ranks of this family, whose members are known as the actin-related proteins (arps), are expanding rapidly . Arps are but one branch of a larger superfamily which includes the actins, hsp/hsc70s, sugar kinases and several cell cycle proteins from bacteria . The existence of the superfamily has been inferred from tertiary structural data . In the case of the arps, their identification and classification has been based upon primary structural data . Placing the arps in a functional context is proving a slower process, although genetic and biochemical analyses are converging in several cases . In the past year, different arps have been linked to functions mediated by actin filaments (arp2 and arp3), microtubules (arp1) and the structural elements of chromatin (arp4 and arp6). J Clin Microbiol, 1996 Feb, 34(2), 324 - 8 Multiprimer PCR system for differential identification of mycobacteria in clinical samples; Del Portillo P et al.; A novel multiprimer PCR method with the potential to identify mycobacteria in clinical samples is presented . The assay relies on the simultaneous amplification of three bacterial DNA genomic fragments by using different sets of oligonucleotide primers . The first set of primers amplifies a 506-bp fragment from the gene for the 32-kDa antigen of Mycobacterium tuberculosis, which is present in most of the species belonging to the genus Mycobacterium . The second set of primers amplifies a 984-bp fragment from the IS6110 insertion sequence of the bacteria belonging to the M . tuberculosis complex . The third set of primers, derived from an M . tuberculosis species-specific sequence named MTP40, amplifies a 396-bp genomic fragment . Thus, while the multiprimer system would render three amplification fragments from the M . tuberculosis genome and two fragments from the Mycobacterium bovis genome, a unique amplification fragment would be obtained from nontuberculous mycobacteria . The results obtained, using reference mycobacterial strains and typed clinical isolates, show that the multiprimer PCR method may be a rapid, sensitive, and specific tool for the differential identification of various mycobacterial strains in a single-step assay. Zhonghua Liu Xing Bing Xue Za Zhi, 1996 Feb, 17(1), 44 - 6 {A comparison of purified urease antigen and whole cell antigen of Helicobacter pylori by ELISA test--study on the application and serum diagnoses of Helicobacter pylori urease diagnostic reagent}; Chen JJ et al.; A useful assay for epidemiological survey of H . pylori infection was reported, using the urease antigen of H . pylori to detect the anti-urease antibody in sera from 676 patients suffered from gastropathy with ELISA technique, and compared with whole cell antigen . Results showed that the purified urease antigen was better than whole cell antigen . The partially purified urease antigens rapid diagnostic reagent of H . pylori was examined in hospital/institution and compared with whole-cell antigens . Results of sera from 676 H . pylori-positive gastritis and non-ulcer dyspepsia patients being tested showed that specificity and sensitivity of ELISA were 96% and 98% respectively . It seemed to be very useful for epidemiological studies on H . pylori infection . The use of ELISA in the detection of IgG antibodies against H . pylori was also sensitive, specific and rapid in assessing the improvement of both acute and chronic inflammation, cleaning of bacteria and the antibody titers after treatment, so as recognized an ideal diagnostic method. Phys Med Biol, 1996 Feb, 41(2), 205 - 22 From targets to genes: a brief history of radiosensitivity; Steel GG; The biological work of Douglas Lea spanned the period from 1934 to his early death in 1947, and during this short period he made important contributions to the theory of radiation action . He interpreted experimental data relating to the effects of radiation on viruses, bacteria, bean roots, etc in terms of the inactivation of discrete targets, which he identified with cellular genes . He thus laid the foundation of much subsequent research . It is now well recognized that mammalian cells differ substantially in radiosensitivity, especially in the low-dose region of the survival curve . The dependence of radiosensitivity on dose rate has been widely studied; this has practical significance for clinical radiotherapy as well as mechanistic implications . Since Lea's time there have been a number of efforts to describe models that can relate cell killing to radiation dose, dose rate, and track structure . So far these have not led to a comprehensive and widely accepted picture . Microdosimetric considerations lead to the concept of differing severity of lesions induced in DNA . Much is known about the sequence of processes that subsequently lead to cell inactivation: this can be divided into phases of induction, processing, and manifestation . Chromosomal events are currently attracting much attention, as they did in Lea's time . Considerable progress has also been made in identifying genes that control the repair of radiation damage . It has been found that mutation is frequently associated with the loss of a large segment of the genome around the damage site and this will have important implications for interactive processes between particle tracks. Int J Dev Biol, 1996 Feb, 40(1), 323 - 30 Signals and signal-transduction systems in the control of development in Hydra and Hydractinia; Hassel M et al.; Pattern control in Hydra has traditionally been assigned to the determining influence of morphogens and neuropeptides . However, at present, arachidonic acid and its derivative 12-S-HETE are the only identified, potential signal molecules known to promote head and bud formation . More potent factors might exist but are not yet identified . Nonetheless, it is possible to evoke the development of an almost unlimited number of supernumerary head structures and to induce ectopic foot formation by interference with the PI-PKC signal transducing system . Such an interference can also rescue the regeneration-deficient mutant reg-16 . Regarding signals in the development of Hydractinia, metamorphosis is induced by an external key stimulus, i.e . a lipid derived from environmental bacteria . The reception of this stimulus involves PKC-mediated responses . Upon its reception, a neuropeptide is released as an internal, synchronising signal . Members of the novel LWamide family of peptides appear to represent this internal signal . In postmetamorphic development, a glycoprotein SIF serves as an inducer of stolon formation. Bull Environ Contam Toxicol, 1996 Feb, 56(2), 279 - 86 Assessment of the impact of naphthalene contamination on mangrove fauna using behavioral bioassays; Mackey AP et al.; Pollution of the marine and estuarine environments by petroleum hydrocarbons is a world wide phenomenon (Connell and Miller 1980) and whilst large scale crude oil spills are the most obvious source of pollution, since the 1970's the impact of chronic, low level hydrocarbon input from sources such as oil refineries has been recognised as having long term ecological consequences, even when there may be no visible evidence of acute effects (Connell and Miller 1980) . Mangroves are perhaps the dominant and most important intertidal habitat along subtropical and tropical coastlines and estuaries and as such are located in areas of high risk of acute or chronic petroleum hydrocarbon pollution . Further, once contamination occurs, high levels of hydrocarbons may be expected to remain in mangrove sediments as conditions are not favorable for hydrocarbon depletion by sediment transport or degradation by aerobic bacteria . Much research has focused on determining the acute toxicity of the water soluble fraction of crude or fuel oil to aquatic fauna but relatively little attention has been given to individual hydrocarbons . The medium to low boiling point aromatics such as naphthalene and its alkyl derivatives are the most toxic petroleum fraction to marine organisms (Anderson et al . 1974; Moore and Dwyer 1974) and they are known to provoke behavioral responses in marine animals at sublethal concentrations (Hargrave and Newcomb 1973; Linden 1977; Nagarajah et al . 1985) . The goal of this investigation was to investigate the effects of a single aromatic petroleum hydrocarbon, naphthalene, in a subtropical mangrove environment, through the use of behavioral bioassays . The test organism chosen was the intertidal gastropod Ophicardelus quoyi, which is abundant in mangroves throughout eastern Australia. Zhonghua Hu Li Za Zhi, 1996 Feb, 31(2), 68 - 9 {The effect of air purification on 10,000 degree clean room in supply rooms}; Yang YL et al.; This article introduced that purifying equipments were installed in the major operating departments of supply rooms in hospital, and that experimental tests were done on the packing department . In statil state, the air humidity and the number of bacteria in the room after purification was obviously lower than that before purification (P < 0.01) . In trending conditions, the main technical index on 10,000 degree clean rooms were examined, all of the results were up to the national standards. Thorax, 1996 Feb, 51(2), 150 - 4 Induction of interleukin 6 and interleukin 8 expression by Broncho-Vaxom (OM-85 BV) via C-Fos/serum responsive element; Keul R et al.; BACKGROUND: Broncho-Vaxom (OM-85 BV) increases the resistance of the respiratory tract to bacterial infections by modulating host immune responses . The compound increases serum IgG levels but decreases IgE levels in patients suffering from chronic bronchitis or chronic obstructive pulmonary disease . It increases concentrations of gamma-interferon (IFN-gamma), IgA, and interleukin (IL)-2 in bronchoalveolar lavage fluid of patients with bronchitis . Treatment with OM-85 BV increases the number of T helper and natural killer cells . In this study the effects of OM-85 BV on transcription of cytokines is investigated in human lung fibroblasts . METHODS: Transcription and synthesis of IL-6 and IL-8 were assessed in cultured primary human lung fibroblasts using standard methods of Northern blot analysis for the level of mRNAs and enzyme linked immunosorbent assay for proteins . RESULTS: Broncho-Vaxom (OM-85 BV) at different concentrations induced transcription of IL-6 and IL-8 . The effect of the drug on transcription of IL-6 and IL-8 genes correlated with secretion of the proteins into cell supernatants . OM-85 BV-dependent expression of the interleukin genes involved C-Fos/serum responsive element (C-Fos/SRE) . CONCLUSIONS: The data suggest that the various immunopharmacological activities of OM-85 BV that have been described in clinical studies may be explained by its ability to induce expression of IL-6 and IL-8. J Heart Lung Transplant, 1996 Feb, 15(2), 196 - 205 Diagnostic yield and therapeutic impact of flexible bronchoscopy in lung transplant recipients; Chan CC et al.; BACKGROUND: Bronchoalveolar lavage and transbronchial biopsy are often used for definitive diagnosis of lung rejection and infection in lung transplant recipients . Although protected specimen brushing is of value in nosocomial bacterial pneumonia, its role in lung transplant recipients had not been widely reported . The aim of the study is to review the diagnostic yield and therapeutic impact of flexible bronchoscopy with the use of a combination of bronchoalveolar lavage, protected specimen brushing, and transbronchial biopsy in lung transplant recipients . METHODS: We reviewed flexible bronchoscopy data in 83 transplant recipients between February 1990 and March 1995 . Only those with bronchoalveolar lavage, protected specimen brushing, and transbronchial biopsy were included in the analysis . There were 282 bronchoscopies performed for clinically suspected lung rejection or infection (clinical bronchoscopy) and 38 bronchoscopies for follow-up of a previously detected histologic abnormality (follow-up bronchoscopy) . RESULTS: The total yields for rejection and infection for clinical and follow-up bronchoscopies were 67.4% and 58.9%, respectively . Acute rejection was detected with transbronchial biopsy in 26.2% and 34.2% of clinical and follow-up bronchoscopies, respectively . Cytomegalovirus pneumonitis was detected with transbronchial biopsy in 4.0% and 11.4% of clinical and follow-up bronchoscopies, respectively . Overall, bacteria was the most common cause of lower respiratory tract infection . When used together, protected specimen brushing and bronchoalveolar lavage were complementary techniques for detection of bacterial lower respiratory tract infection with a significantly higher proportion detected with protected specimen brushing ( > or = 10(3) colony forming units/ml) compared with bronchoalveolar lavage ( > or = 10(5) colony forming units/ml) (p < 0.001) . Complications were hemorrhage (1.9%), pneumothorax (2.5%) and transient hypoxemia (10.5%) . The results had an impact on management of rejection and infection in 57.8% of clinical and 39.5% of follow-up bronchoscopies . CONCLUSIONS: We conclude that bronchoscopy, with the use of a combination of bronchoalveolar lavage, protected specimen brushing, and transbronchial biopsy, is safe with a high diagnostic yield and therapeutic impact for treating lung transplant recipients. Curr Eye Res, 1996 Feb, 15(2), 151 - 5 Binding of Acanthamoeba to hydrogel contact lenses; Gorlin AI et al.; The numbers of Acanthamoeba binding to new hydrogel contact lenses of different polymer and water content were determined with two quantitative methods, a radiolabeled-cell method and a detaching-fluid method . Numbers of amoebae retained on nonionic lenses increased with increasing water content of the lenses . With both nonionic and ionic lenses numbers of associated amoebae decreased with successive rinsing steps . The retentions of amoebae on unworn hydrogel lenses, in contrast to the irreversible adhesion of bacteria, were tenuous and appeared to be effected mainly by surface tension, surface charge and water content. J Hosp Infect, 1996 Feb, 32(2), 99 - 104 Contamination of central venous catheters . The skin insertion wound is a major source of contamination; Egebo K et al.; In a prospective controlled trial we compared the rates of catheter-tip contamination in central venous catheters inserted with or without skin contact . The study was designed so that each patient was their own control . All patients had a single-lumen central venous catheter and a Swan-Gantz sheet inserted through the skin . A Swan-Gantz catheter was inserted and retracted through the sheet thus avoiding contact with skin or subcutaneous tissue . Catheter-tip cultures were performed on removal of catheters . Thirty-three Swan-Gantz catheters were cultured and all were sterile . In the corresponding 33 sheets 16 (48.6%) yielded bacterial growth . Four of the sheets showed growth of more than 15 cfu . In the 26 single-lumen catheters, eight (30.8%) catheter-tips grew bacteria, and four of them had more than 15 colonies . The study supports the theory that the skin-insertion wound is a major source of catheter-contamination. Aust Vet J, 1996 Feb, 73(2), 45 - 9 Effects of posture and accumulated airway secretions on tracheal mucociliary transport in the horse; Raidal SL et al.; Tracheal mucociliary clearance was determined in horses by measuring the rostrad transport of the radiopharmaceutical 99mtechnetium-sulphur colloid following deposition on the tracheal epithelium by intratracheal injection . The effects of head position (head elevated to normal standing position vs head lowered) and of accumulated purulent secretions on tracheal mucociliary clearance were evaluated for the first time in the horse . In normal horses tracheal mucociliary clearance was greatly accelerated by lowering the head so that the cranial trachea was lower than the caudal trachea . Horses confined with their heads elevated for 24 hours developed an accumulation of purulent airway secretions (and associated increased numbers of bacteria) in the lower respiratory tract and showed a decrease in tracheal mucociliary clearance when compared with their previously measured rate when the lower airway contained only normal secretions . These findings have implications for management practices where horses are prevented from lowering their heads, such as transportation and cross-tying, which may therefore contribute to lower respiratory tract disease in horses. Z Gastroenterol, 1996 Feb, 34(2), 116 - 22 Helicobacter pylori produces histamine and spermidine; Velasquez RD et al.; The mechanism by which Helicobacter pylori associates with the development of upper gastrointestinal diseases is still not well understood . Toxic metabolites of H . pylori are discussed as possible factors . We were interested to investigate, whether biogenic amines might be involved . Ten monocultures of H . pylori from the antrum of patients with H . pylori associated diseases were analyzed for the content of the biogenic amine histamine . The bacteria were isolated on Columbia blood agar with Skirrow's supplement in a microaerophilic environment . After three passages onto fresh agar plates the bacteria were harvested in their sonicated suspensions analyzed by reversed-phase high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) . From the HPLC analysis, histamine was found in six cultures . Analysis with GC-MS, however, revealed the presence of histamine in all the cultures (0.1-1.63 nmol histamine/10(8) bacteria) . Additionally to histamine, all cultures were found to contain spermidine in concentrations of 0.010-7.912 nmol/10(8) bacteria . It is discussed that histamine produced by H . pylori may be involved in the pathogenesis of H . pylori associated gastrointestinal diseases. Mod Pathol, 1996 Feb, 9(2), 126 - 32 Post-lung transplant biopsies: an 8-year Loyola experience; Husain AN et al.; A total of 125 transplant procedures involving the lung have been performed at Loyola University of Chicago in 120 patients . There were 67 single (40 right, 27 left), 44 bilateral single, 2 double lung, and 12 heart-lungs (HL) transplant procedures . This paper summarizes the pathologic findings in 565 transbronchial, 102 endobronchial, 20 open lung, and 92 endomyocardial biopsies and compares them with the recommendations in the published literature . The lung biopsies were evaluated according to the Working Formulation, Lung Rejection Study Group, International Society of Heart Transplantation . In transbronchial biopsies, all of which were from the transplanted lungs, the number of alveolated lung fragments ranged from 0 to 14 (mean, 5) . Two hundred twelve biopsies showed no rejection, 113 had minimal rejection, 133 had mild rejection, 34 had moderate rejection, and 1 had severe acute rejection . Active airway damage (Grade B) was seen in 48 biopsies, which were graded from minimal to severe based on the amount of inflammation . Chronic rejection (Grade C) was diagnosed in 23, chronic vascular rejection (Grade D) in 8, and acute vasculitis (Grade E) in 9 biopsies . Routine trichrome and elastic van Gieson stains did not add to the diagnosis . All biopsies were routinely stained with immunoperoxidase for cytomegalovirus . Cytomegalovirus was diagnosed in 84 biopsies, 54 by both H&E and immunoperoxidase, 23 by immunoperoxidase alone, and 5 by H&E alone . The endobronchial biopsy of the anastomotic site had nonspecific inflammation in 46 biopsies . Twenty-nine had infection with a specific organism, Aspergillus and Candida in each of 8 biopsies by Gomori's methenamine silver stain, cytomegalovirus in 7 (4 by H&E and immunoperoxidase; 3 by immunoperoxidase), bacteria in 4, and fungal hyphae in 2 biopsies . In the 12 patients with heart-lung transplants, a total of 92 endomyocardial, 35 transbronchial, and 1 endobronchial biopsies were obtained . Acute rejection was seen only in 2 endomyocardial biopsies, whereas the transbronchial biopsy showed acute mild or moderate rejection in 10, chronic rejection in 1, and cytomegalovirus infection in six biopsies . We conclude that: (a) all biopsies with alveolated lung parenchyma can be evaluated for rejection and infection yielding clinically significant diagnoses; (b) sections from three levels stained by H&E are essential for evaluation; (c) routine Gomori's methenamine silver, elastic van Gieson, and trichrome stains are not required for transbronchial biopsy, however, routine Gomori's methenamine stain is recommended for all anastomotic site biopsies; (d) routine immunoperoxidase for cytomegalovirus is extremely helpful; (e) Grade B rejection should be further graded; and (f) endomyocardial biopsy played no significant role in the management of heart-lung recipients. J Clin Pathol, 1996 Feb, 49(2), 176 - 7 Helicobacter pylori infection in patients with Barrett's oesophagus: a prospective immunohistochemical study; Ricaurte O et al.; The prevalence of Helicobacter pylori infection in patients with Barrett's oesophagus was studied prospectively . A sensitive immunohistochemical staining of H pylori was performed in oesophageal and gastric biopsies of 73 patients from a surveillance group with this condition . H pylori was detected in 11 cases of Barrett's mucosa (15%) and in 26 gastric mucosa specimens (35.6%) . All cases positive in Barrett's mucosa were also positive in the stomach . In Barrett's oesophagus, H pylori was never found on specialised epithelium . The percentage of Barrett's mucosa showing inflammatory changes was similar in specimens with and without H pylori, both for chronic (81% v 79%) and acute (9% v 10%) infiltrates . These results indicate that H pylori infection does not play an aetiological role in Barrett's oesophagus and that colonisation of the metaplastic mucosa by this bacteria is related with the presence of gastric type mucosa in the oesophagus and of H pylori infection in the stomach. J Pediatr Gastroenterol Nutr, 1996 Feb, 22(2), 144 - 7 Correlation of hydrogen and methane production to rice carbohydrate malabsorption in Burmese (Myanmar) children; Bolin TD et al.; Rice carbohydrate malabsorption is common in Burmese village children and adults and may contribute to diminished growth . Its diagnosis depends on a rice breath hydrogen test, which has limitations . Almost 20% of Burmese children under age 5 produce methane, compared with less than 7% of children in Africa and Hong Kong . If an increased carbohydrate load in the colon due to rice malabsorption provides increased substrate for methanogenic bacteria in the left colon, higher fasting breath methane concentrations might be a simpler method of diagnosing rice malabsorption . We tested breath hydrogen and methane over a 4-h period and did anthropometric measurements in 142 subjects, 79 children, and 63 adults . Seventy percent of children were rice-malabsorbers . Methane production occurred in 20% of children under 5 years of age and increased to 60% of adults . There is an association of rice malabsorption with reduced length . There was not correlation between rice malabsorption and breath methane, and the concentration of breath methane does not, therefore, indicate rice absorption status and cannot replace rice breath hydrogen tests. Scand J Immunol, 1996 Feb, 43(2), 181 - 92 Intranasal inoculation of Bordetella bronchiseptica in mice induces long-lasting antibody and T-cell mediated immune responses; Gueirard P et al.; Humoral and cellular immune responses were analysed in mice inoculated intranasally with Bordetella bronchiseptica . After infection, the number of bacteria that colonized the respiratory tract of the mice increased during the first day and decreased thereafter . Total IgG levels increased as early as 14 days after infection and decreased with time after infection, whereas total IgA and IgM levels were lower but remained stable . Specific antibodies to the bacteria were mainly IgG2a and IgA and persisted up to 10 months after infection . Some of these specific antibodies were directed against adenylate cyclase-haemolysin, the bacterial factor that had been shown to be necessary for initiation of infection . The proliferation of Bordetella bronchiseptica-reactive spleen cells occurred during the acute phase of infection . T cells from infected mice produced increasing amounts of IFN gamma and IL-2 after infection . Although very low levels of IL-10 were produced, no IL-4 was detected after bacterial stimulation in vitro . These results suggest that Bordetella bronchiseptica infection induces primarily a Th1-type T-cell response . Importantly, the authors demonstrated that antibody and T-cell responses directed against bacterial determinants of the virulent strain and to purified adenylate cyclase-haemolysin were long-lasting . This observation could be due to the fact that Bordetella bronchiseptica may persist intracellularly in the host as it was demonstrated in vitro. Endod Dent Traumatol, 1996 Feb, 12(1), 33 - 7 Effect of various sealers with gutta-percha as root-end fillings on healing after replantation; Pitt Ford TR et al.; This study examined histologically the effect of three sealers used with gutta-percha as root-end fillings placed in a replantation model . A total of 14 molar teeth were used in seven monkeys . After extraction, root ends were resected, the canals contaminated with oral bacteria, root-end cavities prepared, and fillings of gutta-percha and sealer placed prior to replantation . After 8 weeks the teeth and surrounding jaw were removed and prepared for histological examination . Six roots filled with gutta-percha plus a fortified zinc oxide-eugenol cement (IRM), 13 with gutta-percha plus zinc oxide-eugenol (EP), and eight with gutta-percha plus Kloropercha, were available for examination . The tissue response to gutta-percha with either zinc oxide material was characterized by little or no inflammation of limited extent . In contrast, more severe inflammation was observed against root ends filled with gutta-percha and Kloropercha . Gutta-percha plus IRM or zinc oxide-eugenol had a much more favourable response than that to amalgam reported previously in a similar experiment. Clin Immunol Immunopathol, 1996 Feb, 78(2), 172 - 9 Cultured Kaposi's sarcoma tumor cells fail to stimulate T cell proliferation; Foreman KE et al.; Prior to the AIDS epidemic, Kaposi's sarcoma (KS) was a rare neoplasm . However, in the context of immunosuppression, cutaneous KS lesions more frequently develop and express various surface molecules recognized by T cells such as intercellular adhesion molecule-1 (ICAM-1; CD54) and HLA-DR . The KS tumor cells are thought to arise locally from endothelial cells via a transdifferentiation process . To determine if KS tumor cells can stimulate resting T cell proliferation, we asked whether the tumor cells express the critically important T cell costimulatory molecules B7-1 (CD-80) and B7-2 (CD-86) . In contrast to cytokine-activated endothelial cells, which were induced to express B7-1, but not B7-2 and could function in bacteria-derived superantigen-driven T cell proliferation, four different KS tumor cell lines failed to express either B7-1 or B7-2 and were unable to stimulate allogeneic T cell proliferation upon addition of bacteria-derived superantigen . These results suggest that KS tumor cells behave differently in their response to cytokines compared with endothelial cells and may be able to evade the local immune response by not expressing costimulatory molecules necessary for T cell proliferation. Virology, 1996 Feb 1, 216(1), 184 - 96 The novel protein kinase of the RR1 subunit of herpes simplex virus has autophosphorylation and transphosphorylation activity that differs in its ATP requirements for HSV-1 and HSV-2; Peng T et al.; The large subunit of herpes simplex virus (HSV) ribonucleotide reductase (RR1) designated ICP6 and ICP10 for HSV-1 and HSV-2, respectively, has a novel protein kinase (PK) enzymatic activity . ICP10 is localized on the cell surface, a localization that depends on an intact transmembrane (TM) segment . We used immunocomplex PK assays to examine the PK activity of ICP10 in stably transfected eukaryotic cells . Activity was distinct from that of casein kinase II (CKII) in that it did not require monovalent ions and was not inhibited by zinc sulfate . PK activity was eliminated by deletion of the conserved PK catalytic motifs or of the TM segment and it was significantly impaired by mutation of the invariant Lys (Lys176) . Loss of PK activity by Lys176 mutation resulted in the failure to bind ATP . A truncated ICP10 PK expressed in bacteria (pp29 1a1) retained auto- and transphosphorylating activity (for calmodulin) after purification to apparent homogeneity . PK activity was also absent in cells infected with a recombinant virus (ICP10 delta PK) deleted in the ICP10 PK catalytic motifs . In cells infected with HSV-1 or HSV-2, RR1 had auto- and transphosphorylating activity for the small subunit of HSV ribonucleotide reductase (RR2) and immunoglobulin G (IgG) . Comparing the PK activity of ICP6 and ICP10 we found that ICP6 requires five-fold higher concentrations of {gamma-32P}ATP than ICP10 and both enzymes are Mn2+ dependent, which is also different from CKII that is primarily Mg2+-dependent . Similar results were obtained for various HSV strains and in different cell lines . The data are consistent with the conclusion that the RR1 PK activity is intrinsic. Oral Microbiol Immunol, 1996 Feb, 11(1), 42 - 50 Prevotella intermedia fimbriae mediate hemagglutination; Leung KP et al.; Our earlier studies demonstrated that clinical strains of Prevotella intermedia, isolated from human periodontal pockets, possess various types of fimbria (surface appendages) as determined ultrastructurally . These bacteria have the ability to agglutinate selected mammalian erythrocytes . Hemagglutinating activity exhibited by these cells may be attributable to these surface structures . Strain 17, which possess fimbriae of 8 nm in diameter and readily agglutinates human, monkey, sheep, rabbit, and mouse erythrocytes was selected to determine whether these fimbriae possessed the hemagglutinating activity . Fimbriae were mechanically sheared, concentrated by ammonium sulfate precipitation, solubilized in 10 mM Tris-HCl buffer containing 0.5% deoxycholate and partly purified by ultracentrifugation in a 10-50% linear sucrose gradient . Isolated fimbriae banded at a density of 1.20-1.15 g/ml, appeared fairly uniform ultrastructurally, and possessed hemagglutinating activity . The hemagglutinating activity of P . intermedia whole cells and isolated fimbriae was reduced by treatment with proteases and eliminated by treatment with heat at 80 degrees C for 10 min . The optimal pH for the hemagglutination was 7.0 . In the process of hemagglutination, P . intermedia whole cells and isolated fimbriae bound to rabbit erythrocytes as observed by: (a) decrease in the hemagglutinating activity of bacterial whole cells and isolated fimbriae after incubation with rabbit erythrocytes, (b) increase in hemagglutinating activity of the red cells used for absorption, and (c) the presence of P . intermedia whole cells and fimbriae on red cells after absorption as revealed by immunoelectron microscopy . Both the immune immunoglobulin G of the rabbit polyclonal anti-fimbriae antibody and the immune immunoglobulin G and its Fab fragments of the murine monoclonal anti-fimbriae antibody were effective inhibitors of hemagglutination mediated by isolated fimbriae . Immunogold labeling showed that the monoclonal antibody bound specifically to P . intermedia fimbriae . These results collectively suggest that the hemagglutination exhibited by P . intermedia may be attributable to the fimbriae or the fimbrial-associated components. Dig Dis Sci, 1996 Feb, 41(2), 263 - 71 Mucus production of choledochal epithelial cells due to bile stagnation in the rat; Kurumado K et al.; Mucus secretion is increased by various causative agents of inflammation in the mucosa . Although mucus production of the choledochal epithelial cells (except goblet cells) has never been noted in the rat in normal physiological conditions, it is seen as an inflammatory reaction in animals with choledochoenterostomy, which brings bacteria and active digestive enzymes into the common bile duct through regurgitation of intestinal contents . It is known that stagnant bile alone can cause inflammation in the mucosa of the gallbladder in patients with aseptic acalculous cholecystitis . In this study, aseptic bile stagnation was caused by choledochal dilation made by detaching the common bile duct from the hepatoduodenal ligament in five rats, and histological changes of the choledochal epithelium were observed by light and transmission electron microscopy 17 months after the operative procedure to determine whether stagnant bile could cause mucus production in choledochal epithelial cells . Mucus production was noted in two rats by light microscopy and in all the rats by transmission electron microscopy, and so it was demonstrated that stagnant bile could give rise to this phenomenon in the rat choledochal epithelium. Mutat Res, 1996 Feb 1, 349(2), 263 - 88 Series: current issues in mutagenesis and carcinogenesis, No . 65 . The genotoxicity and carcinogenicity of paracetamol: a regulatory (re)view; Bergman K et al.; The publication of several studies reporting genotoxic effects of paracetamol, one of the world's most popular over-the-counter drugs, has raised the question of regulatory action . Paracetamol does not cause gene mutations, either in bacteria or in mammalian cells . There are, however, published data giving clear evidence that paracetamol causes chromosomal damage in vitro in mammalian cells at high concentrations and indicating that similar effects occur in vivo at high dosages . Available data point to three possible mechanisms of paracetamol-induced genotoxicity: (1) inhibition of ribonucleotide reductase; (2) increase in cytosolic and intranuclear Ca2+ levels; (3) DNA damage caused by NAPQI after glutathione depletion . All mechanisms involve dose thresholds . Studies of the relationship between genotoxicity and toxic effects in the rat (induction of micronuclei in rat bone marrow including dose-response relationship, biotransformation of paracetamol at different dosages, concomitant toxicity and biochemical markers) have recently been completed . These studies, which employed doses ranging from the dose resulting in human therapeutic peak plasma levels to highly toxic doses, give convincing evidence that genotoxic effects of paracetamol appear only at dosages inducing pronounced liver and bone marrow toxicity and that the threshold level for genotoxicity is not reached at therapeutic dosage . Reliable studies on the ability of paracetamol to affect germ cell DNA are not available . However, based on the amount of drug likely to reach germ cells and the evidence of thresholds, paracetamol is not expected to cause heritable damage in man . Various old and poorly designed long-term studies of paracetamol in the mouse and rat have given equivocal results . A few of these studies showed increased incidence of liver and bladder tumours at hepatotoxic doses . National Toxicology Program (U.S.A.) feeding studies have shown that paracetamol is non-carcinogenic when given at non-hepatotoxic doses up to 300 mg/kg/d to the rat and up to 1000 mg/kg/d to the mouse . Taking into account the knowledge of the hepatotoxicity and metabolism of paracetamol and the existence of thresholds for its genotoxicity, the animal studies do not indicate a carcinogenic potential at non-hepatotoxic dose levels . Based on this updated assessment of the genotoxicity and carcinogenicity of paracetamol, it is concluded that there is no need for regulatory action. Appl Environ Microbiol, 1996 Feb, 62(2), 668 - 75 Isolation and identification of methanogen-specific DNA from blanket bog peat by PCR amplification and sequence analysis; Hales BA et al.; The presence of methanogenic bacteria was assessed in peat and soil cores taken from upland moors . The sampling area was largely covered by blanket bog peat together with small areas of red-brown limestone and peaty gley . A 30-cm-deep core of each soil type was taken, and DNA was extracted from 5-cm transverse sections . Purified DNA was subjected to PCR amplification with primers IAf and 1100Ar, which specifically amplify 1.1 kb of the archaeal 16S rRNA gene, and ME1 and ME2, which were designed to amplify a 0.75-kb region of the alpha-subunit gene for methyl coenzyme M reductase (MCR) . Amplification with both primer pairs was obtained only with DNA extracted from the two deepest sections of the blanket bog peat core . This is consistent with the notion that anaerobiosis is required for activity and survival of the methanogen population . PCR products from both amplifications were cloned, and the resulting transformants were screened with specific oligonucleotide probes internal to the MCR or archaeal 16S rRNA PCR product . Plasmid DNA was extracted from probe-positive clones of both types and the insert was sequenced . The DNA sequences of 8 MCR clones were identical, as were those of 16 of the 17 16S rRNA clones . One clone showed marked variation from the remainder in specific regions of the sequence . From a comparison of these two different 16S rRNA sequences, an oligonucleotide was synthesized that was 100% homologous to a sequence region of the first 16 clones but had six mismatches with the variant . This probe was used to screen primary populations of PCR clones, and all of those that were probe negative were checked for the presence of inserts, which were then sequenced . By using this strategy, further novel methanogen 16S rRNA variants were identified and analyzed . The sequences recovered from the peat formed two clusters on the end of long branches within the methanogen radiation that are distinct from each other . These cannot be placed directly with sequences from any cultured taxa for which sequence information is available. Mol Cell Biol, 1996 Feb, 16(2), 593 - 602 Identification of human proteins functionally conserved with the yeast putative adaptors ADA2 and GCN5; Candau R et al.; Transcriptional adaptor proteins are required for full function of higher eukaryotic acidic activators in the yeast Saccharomyces cerevisiae, suggesting that this pathway of activation is evolutionarily conserved . Consistent with this view, we have identified possible human homologs of yeast ADA2 (yADA2) and yeast GCN5 (yGCN5), components of a putative adaptor complex . While there is overall sequence similarity between the yeast and human proteins, perhaps more significant is conservation of key sequence features with other known adaptors . We show several functional similarities between the human and yeast adaptors . First, as shown for yADA2 and yGCN5, human ADA2 (hADA2) and human GCN5 (hGCN5) interacted in vivo in a yeast two-hybrid assay . Moreover, hGCN5 interacted with yADA2 in this assay, suggesting that the human proteins form similar complexes . Second, both yADA2 and hADA2 contain cryptic activation domains . Third, hGCN5 and yGCN5 had similar stabilizing effects on yADA2 in vivo . Furthermore, the region of yADA2 that interacted with yGCN5 mapped to the amino terminus of yADA2, which is highly conserved in hADA2 . Most striking, is the behavior of the human proteins in human cells . First, GAL4-hADA2 activated transcription in HeLa cells, and second, either hADA2 or hGCN5 augmented GAL4-VP16 activation . These data indicated that the human proteins correspond to functional homologs of the yeast adaptors, suggesting that these cofactors play a key role in transcriptional activation. J Bacteriol, 1996 Feb, 178(3), 881 - 7 Induction of the gap-pgk operon encoding glyceraldehyde-3-phosphate dehydrogenase and 3-phosphoglycerate kinase of Xanthobacter flavus requires the LysR-type transcriptional activator CbbR; Meijer WG et al.; In a previous study, a gene (pgk) encoding phosphoglycerate kinase was isolated from a genomic library of Xanthobacter flavus . Although this gene is essential for autotrophic growth, it is not located within the cbb operon encoding other Calvin cycle enzymes . An analysis of the nucleotide sequence upstream from pgk showed the presence of a gene encoding glyceraldehyde-3-phosphate dehydrogenase and the 3' end of an open reading frame encoding a protein which is 50% identical to transketolase encoded by cbbT of X . flavus . Gene fusions between pgk and lacZ demonstrated that the gap and pgk genes are organized in an operon . Induction of the Calvin cycle in heterotrophically growing cells resulted in a sixfold increase in phosphoglycerate kinase activity in parallel with the appearance of ribulosebisphosphate carboxylase activity . This superinduction of phosphoglycerate kinase did not occur in an X . flavus strain in which cbbR, encoding the transcriptional activator of the cbb operon, was disrupted . The failure to superinduce the gap-pgk operon is not caused by the absence of a functional Calvin cycle, since the expression of this operon in an X . flavus strain with a defective ribulosebisphosphate carboxylase enzyme was the same as the expression in the wild type . It is therefore concluded that the expression of both the cbb and gap-pgk operons is controlled by CbbR. J Chromatogr A, 1996 Jan 26, 722(1-2), 233 - 43 Determining the characteristics of water pollutants by neural sensors and pattern recognition methods; Yatsenko V; In this paper we have researched the influence of pollutants on such biological objects as photosynthesizing systems in order to reveal the capabilities and features of their application as the controlling sensor in integral ecological monitoring microsystems . It is proposed to elaborate upon the intelligent sensor on the basis of: (1) neural network technologies; (2) the possibility to separate the characteristics of the substances dissolved in water by means of the methods which recognize patterns in a functional space of the fluorescence curves; (3) the results of the chromatographic analysis of standard water samples . This sensor allows to predict water state and to make the optimal decisions for correcting an ecosystem's condition . The efficiency of such a system for water analysis can be improved using the dual measurement principle . This principle suggests identification of a biosensor model according to experimental data. J Biotechnol, 1996 Jan 26, 44(1-3), 171 - 81 Induction of protective immunity by aerosol or oral application of candidate vaccines in a dose-controlled pig aerosol infection model; Hensel A et al.; In order to outline basic concepts for the design of a bacterial aerosol infection model, the development of a pig model with Actinobacillus pleuropneumoniae is described . First, reproducibility of aerosol parameters should be maintained by optimizing generating and sampling conditions . Survival rates of the chosen strain must be predictable . Secondly, inhalation conditions for the recipients have to be standardized to enable the determination of deposition sites and the dose administered . Subsequently, dose-response relationship should be evaluated to find a suitable challenge dose . Furthermore, it seems necessary to establish methods to obtain local specimens for determination of the local immune responses . The present study demonstrates that after aerosol challenge pigs were completely protected after inhalation and partially protected after oral application of A . pleuropneumoniae vaccines and describes techniques to administer bacteria in a dose-dependent, viable way . Using the infection model several stages of the disease from acute pleuropneumonia to chronic infection can be induced for research purposes. J Biotechnol, 1996 Jan 26, 44(1-3), 5 - 11 Antigen handling in the gastrointestinal lamina propria; Stokes CR et al.; Enteric infections associated with bacteria, viruses and parasites are a world health problem accounting for at least one billion episodes of diarrhoea and up to 10 million deaths in the Third World each year . The widely held conclusion that responses which are protective in the gastrointestinal tract are best stimulated by mucosal presentation of antigen remains largely unchallenged . Despite this there are but a few effective mucosal vaccines . The mucosal immune system of the gastrointestinal tract is presented with a contrasting array of antigens, ranging from harmless dietary components to highly pathogenic organisms, and has evolved a battery of responses from which an appropriate response may be orchestrated . We have postulated that the microenvironment in which an antigen is processed and presented to the mucosal immune system may profoundly influence the nature of the response and the purpose of this review is discuss a possible role for the lamina propria in this process. Biochemistry, 1996 Jan 23, 35(3), 698 - 703 Proline pipe helix: structure of the tus proline repeat determined by 1H NMR; Butcher DJ et al.; The structure of a 22 amino acid peptide, TPPI {Nedved, M . L., Gottlieb, P . A., & Moe, G . R . (1994) Nucleic Acids Res . 22, 5024-5030}, that is similar to the proline repeat segment of the replication arrest protein, Tus, has been determined by 1H NMR in 50% trifluroethanol . The structure is a novel left-handed helix having 5.56 residues per turn and a regular hydrogen bonding network that is limited to one side of the helix and contains a channel that runs down the helix axis . The latter feature gives the structure an overall pipe-like appearance; hence, the structure has been designated a proline pipe helix . The Tus proline pipe is also amphiphilic with one side consisting of proline and other nonpolar residues while the other side contains mostly basic and other polar residues . Tus and several other proteins that contain a similar proline repeat sequence are DNA binding proteins . It is shown here that the proline pipe helix of TPPI can be accommodated within the major grove of B-form DNA in a manner that positions nearly all of the basic residues near phosphate groups in the DNA backbone . The proline pipe helical motif may be a structural element of many other proteins including integral membrane receptor proteins. Toxicology, 1996 Jan 22, 107(1), 9 - 15 2,4-Dichlorophenoxyacetic acid affects the attachment of Azospirillum brasilense Cd to maize roots; Jofre E et al.; 2.4-Dichlorophenoxyacetic acid (2,4-D) is a herbicide widely applied to forage, grain and cereals . We previously determined that 1 mM 2,4-D diminished cell growth and cellular activity of Azospirillum brasilense Cd . The present work was designed to determine the possible effect of this herbicide--at concentrations used on crops--on the attachment of the bacteria to maize roots, since this step is of prime importance for the growth stimulation of the plant obtained with Azospirillum brasilense . In this paper we demonstrate that 2,4-D alters the bacterial adhesion to maize roots. Arch Intern Med, 1996 Jan 22, 156(2), 137 - 42 The interface between research and the diagnosis of an emerging tick-borne disease, human ehrlichiosis due to Ehrlichia chaffeensis; Dawson JE et al.; Two new ehrlichial species that cause human disease have recently been identified: Ehrlichia chaffeensis and the currently unnamed agent of human granulocytic ehrlichiosis . Our objective was to review data on the clinical presentation, laboratory and epidemiological findings, therapy, and diagnostic procedures of patients with human ehrlichiosis due to E chaffeensis . From 1986 through 1994, 400 case patients were identified from 30 US states . Most patients had a nonspecific illness, characterized by fever and headache . Severe illness and death occurred, primarily in the elderly . Laboratory findings most commonly included leukopenia, thrombocytopenia, and elevated liver function test results . Antibody response was the basis for diagnosis, although polymerase chain reaction testing has been useful in research settings . Empirical treatment with tetracycline or its analogues should be begun as soon as possible after the onset of symptoms . Clinicians need to be alert for this illness when evaluating febrile patients whose history includes possible recent tick exposure. AIDS Res Hum Retroviruses, 1996 Jan 20, 12(2), 99 - 106 Compositional similarities between the human immunodeficiency virus and surface antigens of pathogens; Fitzgerald DJ et al.; The genome of the human immunodeficiency virus (HIV) is rich in A but not U and deficient in C but not G . This asymmetric nucleotide bias is the major factor in determining the unusual composition of HIV proteins . In this report, we have identified the cellular genes in the GenBank database that are compositionally similar to HIV in order to further understand the significance of the nucleotide bias of the viral genome . A total of 101 genes in the bacterial and invertebrate subdivisions of the database were found to have a base composition that is similar to the composition of the HIV genome . The identified cellular sequences represent a discrete subset of the database since 81 of the 101 entries code for antigens from pathogens and nearly all of these organisms infect humans . The amino acid compositions of these surface antigens are also similar to the unusual composition of HIV proteins, which are deficient in proline and rich in lysine and other polar residues encoded by A-rich codons . The similarities between the HIV proteins and the immunodominant antigens from other pathogens may indicate a common pathogenic strategy for the promotion of immune dysregulation. Vet Rec, 1996 Jan 20, 138(3), 53 - 8 Feline tuberculosis: a literature review and discussion of 19 cases caused by an unusual mycobacterial variant; Gunn-Moore DA et al.; The literature relating to feline mycobacterial disease is reviewed and 19 cats with tuberculosis caused by a previously unknown strain of mycobacterium are discussed . The bacteria were found to have characteristics between those of Mycobacterium tuberculosis and M bovis . The paper considers the clinical signs, epidemiology and diagnosis of the cases, and discusses the possible origins of the organism, treatment regimens and zoonotic potential. J Biol Chem, 1996 Jan 19, 271(3), 1441 - 7 Identification of chitinase as the immunodominant filarial antigen recognized by sera of vaccinated rodents; Adam R et al.; Acanthocheilonema viteae is a parasitic nematode of rodents . We identified the chitinase of A . viteae infective stage larvae (L3) as the main target of the humoral immune response of jirds, which were protected against challenge infection after vaccination with irradiation attenuated L3 . The cDNA of the L3 chitinase has been sequenced, and the deduced amino acid sequence shows significant homologies to chitinases of Brugia malayi microfilariae, insects, yeast, bacteria, and Streptomyces sp . The protein has been characterized by monoclonal antibodies and substrate activity gels . The chitinase of L3 may contribute to degrading the nematode cuticle during molting and thus represents a target of protective immune responses in a phase where the parasite is highly vulnerable . In addition, it has been shown that a similar enzyme exists in uterine microfilariae, which probably has a role in casting the egg shell. Med J Aust, 1996 Jan 15, 164(2), 94 - 5 "Natural" therapy for infectious diseases; Golledge CL et al.; Introduce the right kind of bacteria and several pathogens lose their niche in the human host--that's the rationale behind probiotic therapy . These and some other "natural" therapies are popular with patients and increasingly supported by research results. Genes Dev, 1996 Jan 15, 10(2), 129 - 41 Switching transcription on and off during the yeast cell cycle: Cln/Cdc28 kinases activate bound transcription factor SBF (Swi4/Swi6) at start, whereas Clb/Cdc28 kinases displace it from the promoter in G2; Koch C et al.; When yeast cells reach a critical size in late G1 they simultaneously start budding, initiate DNA synthesis, and activate transcription of a set of genes that includes G1 cyclins CLN1, CLN2, and many DNA synthesis genes . Cell cycle-regulated expression of CLN1, CLN2 genes is attributable to the heteromeric transcription factor complex SBF . SBF is composed of Swi4 and Swi6 and binds to the promoters of CLN1 and CLN2 . Different cyclin-Cdc28 complexes have different effects on late G1-specific transcription . Activation of transcription at the G1/S boundary requires Cdc28 and one of the G1 cyclins Cln1-Cln3, whereas repression of SBF-regulated genes in G2 requires the association of Cdc28 with G2-specific cyclins Clb1-Clb4 . Using in vivo genomic footprinting, we show that SBF (Swi4/Swi6) binding to SCB elements (Swi4/Swi6 cell cycle box) in the CLN2 promoter is cell cycle regulated . SBF binds to the promoter prior to the activation of transcription in late G1, suggesting that Cln/Cdc28 kinase regulates the ability of previously bound SBF to activate transcription . In contrast, SBF dissociates from the CLN2 promoter when transcription is repressed during G2 and M phases, suggesting that Clb1-Clb4 repress SBF activity by inhibiting its DNA-binding activity . Switching transcription on and off by different mechanisms could be important to ensure that Clns are activated only once per cell cycle and could be a conserved feature of cell cycle-regulated transcription. J Immunol, 1996 Jan 15, 156(2), 532 - 8 Ubiquitin-like moiety of the monoclonal nonspecific suppressor factor beta is responsible for its activity; Nakamura M et al.; Monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by murine T cell hybridoma, possesses pleiotrophic Ag-nonspecific suppressive functions . Most recently, a cDNA clone encoding MNSF beta (a subunit of MNSF) was isolated and characterized . The MNSF beta cDNA encodes a 14.5-kDa fusion protein with MNSF-like activity consisting of an 8-kDa ubiquitin-like segment (ubi-L) and the ribosomal protein S30 (6.5 kDa) . To determine whether ubi-L itself has biologic activity, cDNA encoding the ubi-L region was expressed in bacteria and the recombinant product was tested for the activity . Ubi-L showed MNSF-like biologic activity without any cytotoxic action . Interestingly, the addition of ubiquitin to the assay inhibited ubi-L-induced suppression . IFN-gamma, which is known to enhance the expression of MNSF receptor, induced splenocytes to secrete ubi-L by increasing mRNA . Ubi-L has species-specific action and the ability to selectively inhibit the B cell proliferative response stimulated by soluble but not by Sepharose-bound anti-Ig Ab . In addition, okadaic acid, a serine/threonine phosphatase 1 and 2A inhibitor, showed a synergistic inhibitory action with ubi-L, indicative of the possible involvement of phosphatase(s) in the regulation of ubi-L action. J Chromatogr A, 1996 Jan 12, 720(1-2), 3 - 25 High-resolution thin-layer chromatography of gangliosides; Muthing J; In this review an updated overview of current improvements on thin-layer chromatography (TLC) of gangliosides over the past decade is provided . Basic general techniques and special advice is given for successful separation of glycosphingolipids . New approaches concerning continuous and multiple development, and several preparative TLC methods are also included . Emphasis is placed on TLC immunostaining and related techniques, i.e . practical applications of carbohydrate-specific antibodies, toxins and bacteria, viruses, lectins and eukaryotic cells . Thus, this review on ganglioside TLC summarizes its power as an analytical tool for a wide range of purposes. Curr Opin Ophthalmol, 1996 Feb, 7(1), 17 - 20 Anesthesia and preoperative and postoperative medications; Fichman RA; Although certain methods such as retrobulbar blocks are used extensively, improvements in procedure can always be implemented . The use of ultrasound, low concentrations of anesthesia, careful monitoring, and, in the case of risk patients, anesthesia standby are all important considerations to ensure uneventful treatments . Topical anesthesia eliminates needle risk as well as risk of ptosis and bruising . Because it has been demonstrated that bacteria routinely enter the anterior chamber during uncomplicated cataract surgery, certain irrigation solutions are helpful, but still debatable . Postoperatively, diclofenac, flurbiprofen, and timolol have all been proven to be effective in reducing ocular inflammation, reducing incidence of CME, and controlling pressure increase, respectively. Oncogene, 1996 Jan 4, 12(1), 213 - 20 Identification of domains required for transcriptional activation and protein dimerization in the human papillomavirus type-16 E7 protein; Zwerschke W et al.; To analyse the potential of the E7 oncogene of HPV-16 to activate transcription, we constructed hybrid proteins containing various portions of the HPV-16 E7 protein fused to the DNA binding region of the bacterial LexA repressor . We found that full length HPV-16 E7 is capable to mediate activation of two different reporter genes, which carry LexA binding sites in their promoters . In contrast, E7 from HPV-11, a low-risk type papillomavirus, was unable to activate transcription, when analysed in the same assay . Mutations in the transforming domains of HPV-16 E7 did not affect the ability of the protein to activate transcription, indicating that it represents a novel function of the oncoprotein, which is not sensitive to any known inactivating mutations . Analysis of E7 subdomains revealed that the N-terminal part of HPV-16 E7 retains the capacity to activate transcription . A second trans-activation domain is located in the C-terminal part of E7; however, in the context of the full length E7 protein this activity is blocked by an adjacent domain . These results reveal a second pathway for transcriptional activation by HPV-16 E7, independent of its interaction with pRB-E2F complexes . Using the E7-LexA hybrid proteins, it is shown that E7 can form homodimers and this property involves a zinc finger structure in the C-terminal part of the protein, partially overlapping with the domain that negatively regulates transcriptional activation by E7. FEBS Lett, 1996 Jan 2, 378(1), 93 - 7 A sequence analysis of the beta-glucosidase sub-family B; Rojas A et al.; This computational study is a summary of structural properties of the beta-glucosidase subfamily B . Computations were carried out using GCG package programs . All sequences used in this analysis were taken from the protein data bank . The multialignment and the phylogenetic tree of the beta-glucosidase sub-family B are shown . The conserved patterns: DGP, GRNFE, DPYL, KHF, SDW, GLD, VLLKN in the N-terminal region and FGYGLSY in the C-terminal part should be pointed out . C-terminal parts of the Butyrivibrio fibrisolvens and Ruminoccocus albus beta-glucosidase sequences can be aligned to the N-terminal region of the other members of the subfamily . A crossed homology model in sub-family B beta-glucosidases is described. J Exp Biol, 1996, 199(Pt 8), 1743 - 56 Mucus entrapment of particles by a suspension-feeding tilapia (Pisces: Cichlidae) Sanderson S, Stebar M, Ackermann K, Jones S, Batjakas I I, Kaufman L. A miniature fiberoptic endoscope was used to observe the processes of particle encounter and retention inside the buccopharyngeal cavity of suspension-feeding tilapia . Small particles (38 µm to 1.0 mm in diameter) were trapped in strands and aggregates of mucus, which usually slid posteriorly on the ceratobranchials of arches I­IV towards the esophagus while the fish pumped water through the buccopharyngeal cavity . During stage 1 of periodic reversals of water flow inside the buccopharynx, mucus-bound particles usually lifted off the arch surfaces and travelled a short distance in an anterior or anterodorsal direction . During stage 2 of a reversal, the mucus usually resumed travel in a posterior or posteroventral direction and exited the field of view . Mucus was present less often during feeding on large particles (3­10 mm in diameter) than on small particles, and large particles were rarely observed to be attached to mucus . We discuss the advantages to suspension-feeding fishes of using aerosol filtration by mucus entrapment rather than sieving, and predict that many cichlid and cyprinid suspension feeders that consume bacteria and phytoplankton use mucus for aerosol filtration. J Exp Biol, 1996, 199(Pt 6), 1343 - 52 Sulfide may directly modify cytoplasmic hemoglobin deoxygenation in Solemya reidi gills Kraus D, Doeller J, Powell C. The clam Solemya reidi, which survives in sulfide-rich sediments, houses intracellular sulfide-oxidizing bacteria as symbionts in its gills . The gill bacteriocytes also contain a high concentration of cytoplasmic hemoglobin . Although the in situ hemoglobin optical spectrum was not altered in the presence of hydrogen sulfide, hemoglobin deoxygenation was significantly slowed and incomplete when sulfide was present . A sulfide-mediated decrease in oxygen consumption rate, a shift in intracellular pH or the conversion of hemoglobin to an unusual derivative could all slow in situ hemoglobin deoxygenation . However, under low sulfide levels at which deoxygenation is incomplete, oxygen consumption rate was not inhibited, intracellular pH decreased by less than 0.1 units and the only hemoglobin derivatives present were deoxyhemoglobin and oxyhemoglobin . These results and preliminary measurements of the isolated gill hemoglobin dissociation rate constants suggest that sulfide or a rapidly formed oxidation product may directly influence the rate of Solemya reidi gill hemoglobin deoxygenation. Teratog Carcinog Mutagen, 1996, 16(5), 253 - 68 Mechanisms of myricetin mutagenicity in V79 cells: involvement of radicalar species; Silva ID et al.; Myricetin is a flavonol that is widely distributed in edible plants and although it has been proved to be genotoxic in bacteria and to induce significant concentration-dependent nuclear DNA degradation concurrent with lipid peroxidation, very little is known about its mechanisms of genotoxicity . In this work we tried to evaluate the role of rat cytochromes P450 in the genotoxicity of myricetin and to study the role that radicalar species may have in its mutagenicity . The results obtained show that the genotoxicity of myricetin as assessed by the induction of chromosomal aberrations is not different in V79 cells lines genetically engineered for the expression of rat cytochromes P450 1A1, 1A2, and 2B1, compared to parental cell lines . We have also been able to show that reactive oxygen species resulting from the autooxidation of myricetin at pH values above neutrality have an important role in its mutagenicity . Therefore, under some conditions, myricetin can act as a prooxidant. Riv Biol, 1996, 89(1), 139 - 56 A "sensitive" cell system . Its role in a new evolutionary paradigm; Sara M; The cell property of processing the environmental information is called sensitivity and referred to a "sensitive" cell system in which physical signals and specialized molecules work in relation to peculiar cell structures . It is suggested that, in addition to its role in cell behaviour, the sensitive system performs a basic role in morphogenesis in all cells . It interacts with the genetic system in gene expression and regulation directly or indirectly through epigenetic processes . Epigenetic inheritance, adaptive mutations and genetic assimilation allow environmental information to be embodied in new traits, thereby becoming relevant in evolution . This is consistent with an evolutionary mechanism with two alternate and recurrent phases: the first, epigenetic and Lamarckian at cell and organism levels and the second, selective and Darwinian, at population level. Mol Biol Rep, 1996, 23(3-4), 253 - 60 The interaction of C-reactive protein and serum amyloid P component with nuclear antigens; Du Clos TW; The pentraxins are a family of proteins characterized by cyclic pentameric structure, calcium-dependent ligand binding and sequence homology . The two main representatives of this family are the serum proteins, C-reactive protein (CRP) and serum amyloid P component (SAP) . In man CRP is an acute phase reactant which increases up to 1,000 fold during the acute phase response whereas SAP is a constitutive protein expressed at about 30 micrograms/ml . These proteins activate complement through the classical pathway and participate in opsonization of particulate antigens and bacteria . In the past several years it has been determined that both of these pentraxins interact with nuclear antigens including chromatin and small nuclear ribonucleoproteins (snRNPs) . Both CRP and SAP have nuclear transport signals which facilitate their entry into the nuclei of intact cells . Furthermore, these pentraxins have been shown to affect the clearance of nuclear antigens in vivo . It is now believed that one of the major functions of the pentraxins could be to interact with the nuclear antigens released from apoptotic or necrotic cells . This interaction could mitigate against deposition of these antigens in tissue and autoimmune reactivity. Scand J Gastroenterol Suppl, 1996, 221, 18 - 20 Quality of life and Helicobacter pylori eradication; Wilhelmsen I; BACKGROUND: It has convincingly been shown that relapses of duodenal ulcers are significantly reduced after eradication of Helicobacter pylori . The aim of this study was to examine the long-term effect of bacteria eradication on Quality of Life (QOL) and subjective somatic symptoms . METHODS: Seventy-four Helicobacter pylori-positive patients with recurrent duodenal ulcer disease were treated with triple treatment . Relapses, H . pylori status, QOL and subjective somatic symptoms were registered at yearly intervals for 4 years . RESULTS: Preliminary results show that the patients were more satisfied with the healthcare system, had better sexual relationships and less psychological distress up to 3 years after eradication of the bacteria . They also had fewer symptoms of exhaustion and less stomach discomfort than before triple treatment . CONCLUSION: Long-term follow-up of patients after eradication of H . pylori shows that QOL is improved and feeling of exhaustion and stomach discomfort reduced. Langenbecks Arch Chir Suppl Kongressbd, 1996, 113, 1144 - 8 {Surgical prevention of post-traumatic infection by immediate necrectomy of burn wounds}; Pallua N et al.; Sepsis is the commonest cause of death following burn injuries . The main source of the bacteria which cause the onset of sepsis is the burn wound itself . We evaluated the question of whether immediate necrectomy versus early necrectomy leads to a decrease in septic complications, as well as posttraumatic lethality . We evaluated 66 patients, 33 of whom underwent immediate necrectomy (i.e., within the first 3 days posttraumatically, group 1); the remaining 33 patients underwent necrectomy in the early posttraumatic phase (from the fourth posttraumatic day, group 2) . Following immediate necrectomy (group 1), septic complications developed in 12.1%, as compared to 33.3% in group 2 (p < 0.01) . Lathality was significantly reduced in group 1 with 9.1% compared to 21.2% in group 2 (p < 0.01) . In this study it was demonstrated that immediate necrectomy versus early necrectomy in young patients leads to a significant decrease of septic complications and lethality following burn injury. Genes Cells, 1996 Jan, 1(1), 17 - 27 The probabilities of supercoil removal and decatenation by yeast DNA topoisomerase II; Roca J et al.; BACKGROUND: In yeast a single type II DNA topoisomerase is involved in both the removal of DNA supercoils and the unlinking of intertwined pairs of newly replicated chromosomes or plasmids; in bacteria, two type II enzymes, DNA gyrase and DNA topoisomerase IV, function separately in the passage of DNA segments in cis and in trans . To deduce the molecular characteristics of these enzyme-mediated reactions, the efficiencies of supercoil removal and decatenation by the yeast enzyme upon the addition of a nonhydrolysable ATP analogue were determined . RESULTS: The probability that a bound enzyme transports a DNA segment in cis increases with positive or negative supercoiling of the DNA, and transport is nearly quantitative at high degrees of supercoiling . The relative probabilities of transporting a contiguous and noncontiguous DNA segment by a yeast enzyme bound to one member of a singly linked pair of 3.6-kb rings were calculated from the observed efficiency of decatenation . When the enzyme-bound ring is highly supercoiled, transport of a noncontiguous segment is more probable than a contiguous one . CONCLUSION: A DNA-bound yeast enzyme has no intrinsic bias in its selection of a contiguous or noncontiguous DNA segment for transport, and the selection is determined by DNA conformations . For the singly linked dimeric catenane studied, a high degree of supercoiling of the enzyme-bound DNA does not make supercoil removal more favourable than decatenation . In the case of bacterial gyrase, however, wrapping of a DNA segment around the enzyme is expected to strongly favour the transport of a contiguous segment. Postepy Hig Med Dosw, 1996, 50(5), 525 - 8 {Studies of a glycyl component of lipopolysaccharides}; Mieszala M et al.; The aminoacyl analysis of endotoxic lipopolysaccharides (LPS) isolated from several bacteria revealed essential amounts of glycine, among the inherent LPS components . Glycine as a single amino acid was found only in a core part of LPS . The results indicate that amino acid is an integral constituent of core oligosaccharide in lipopolysaccharide. Wien Klin Wochenschr, 1996, 108(21), 669 - 76 {Glutamine: effects on the immune system, protein balance and intestinal functions}; Roth E et al.; Glutamine is the most abundant free amino acid of the human body . In catabolic stress situations such as after operations, trauma and during sepsis the enhanced transport of glutamine to splanchnic organs and to blood cells results in an intracellular depletion of glutamine in skeletal muscle . Glutamine is an important metabolic substrate for cells cultivated under in vitro conditions and is a precursor for purines, pyrimidines and phospholipids . Increasing evidence suggests that glutamine is a crucial substrate for immunocompetent cells . Glutamine depletion in the cultivation medium decreases the mitogen-inducible proliferation of lymphocytes, possibly by arresting the cells in the G0-G1 phase of the cell cycle . Glutamine depletion in lymphocytes prevents the formation of signals necessary for late activation . In monocytes glutamine deprivation downregulates surface antigens responsible for antigen preservation and phagocytosis . Glutamine is a precursor for the synthesis of glutathionine and stimulates the formation of heat-shock proteins . Moreover, there are suggestions that glutamine plays a crucial role in osmotic regulation of cell volume and causes phosphorylation of proteins, both of which may stimulate intracellular protein synthesis . Experimental studies revealed that glutamine deficiency causes a necrotising enterocolitis and increases the mortality of animals subjected to bacterial stress . First clinical studies have demonstrated a decrease in the incidence of infections and a shortening of the hospital stay in patients after bone marrow transplantation by supplementation with glutamine . In critically ill patients parenteral glutamine reduced nitrogen loss and caused a reduction of the mortality rate . In surgical patients glutamine evoked an improvement of several immunological parameters . Moreover, glutamine exerted a trophic effect on the intestinal mucosa, decreased the intestinal permeability and thus may prevent the translocation of bacteria . In conclusion, glutamine is an important metabolic substrate of rapidly proliferating cells, influences the cellular hydration state and has multiple effects on the immune system, on intestinal function and on protein metabolism . In several disease states glutamine may consequently, become an indispensable nutrient, which should be provided exogenously during artificial nutrition. Injury, 1996, 27 Suppl 3, SC27 - 33 Infection after open reduction and internal fixation with dynamic compression plates--clinical and experimental data; Arens S et al.; Infection rates for open reduction and internal fixation (ORIF) with the DCP in clinical studies are based on heterogeneous data on general risk factors and do not take into account the direct effect of the implant (material, design, surface, technique) . The initial degree of bacterial contamination is generally unknown and the applied definition of the term infection is not mentioned . In our own prospective randomized clinical study including 281 cases of ORIF with the DCP (154 steel vs 127 titanium), the influence of the implant material on susceptibility to local infection was examined in relation to initial bacteria contamination (109 non-contaminated / 172 contaminated) . Although in the group of contaminated DCPs the difference in the infection rates for stainless steel (sSt) and commercially pure titanium (cpTi) showed no statistical significance, a tendency was apparent . In an animal experiment, the lower rates of infection for c.p.Ti-DCP compared to sSt-DCP in the presence of a local bacterial challenge could be demonstrated with statistical significance . The need for further experimental research in the field of implant related local infection after ORIF will be discussed and strategies for further investigations proposed. Injury, 1996, 27 Suppl 3, SC5 - 8 Pathophysiology of infection--a theoretical approach; Hansis M; The manifestation of postoperative wound infection has a tri-factorial basis: the overall systemic trauma and the additional effects of premorbidity (age, diabetes, etc.), the local host damage resulting from both the accident and surgery, and the bacterial contamination of the wound . The first factor is only moderately open to intervention, however, the amount of local host damage caused during the operation can be influenced directly by the surgeon who must ensure that his operating techniques are non-aggressive and in line with current knowledge . The factor of the intraoperative bacterial inoculum can be modified by attention to hygiene . The latter two factors are in direct relation to the following two hypotheses: Every wound is able to tolerate some local host damage and some bacterial inoculum without manifestation of infection . The bacterial wound flora is the product of the bacterial invasion force and the local wound conditions . The bacterial wound flora and the local condition of the wound are interrelated . If either factor exceeds the tolerable threshold, infection will become manifest, i.e . there will be an uncontrollable proliferation of bacteria . The level of this breaking point may depend upon certain systemic host factors such as age, diabetes, or immunodeficiency . Consequently, the prevention of infection must focus simultaneously on minimizing the local bacterial inoculum and optimizing local wound conditions . Future studies should concentrate on identifying the exact nature of the individual factors promoting infection, their quantification, and their relative importance. Rom J Morphol Embryol, 1996 Jan-Jun, 42(1-2), 117 - 21 Clinical value of Helicobacter pylori identification by histochemical methods in patients with chronic gastritis; Raica M et al.; There were studied 68 patients admitted with clinical signs of chronic gastritis . Biopsies were taken by endoscopical route from the antral and fundic levels in all cases, and paraffin slides were stained with haematoxylin-eosine, modified Giemsa, Warthin-Starry and blue cresyl methods . Microscopical results were compared with those obtained with the urease test performed from the gastric mucus . Results noted the presence of Helicobacter pylori in 41 from 68 cases in the antral mucosa, like curved bacteria located in the mucus on the excretory segment and often on the surface, covering the epithelium . Intracellular Helicobacter pylori was not noted . Its presence is related especially with the active chronic gastritis and rarely with inactive forms; it was not possible to establish a relationship between the morphology, incidence and features of their infiltrate, and the presence of Helicobacter pylori . The modified Giemsa method is easier to perform, faster and cheaper, and it may be currently applied in practice, but from the sensitivity viewpoint, best results have been achieved with Warthin-Starry silver impregnation. Microbios, 1996, 87(351), 103 - 12 Is the coccoid form of Helicobacter pylori viable? Hua J, Ho B. Helicobacter pylori was grown on solid medium for up to 5 weeks . Morphological conversion from spiral to coccoid forms began after 7 days incubation under microaerophilic conditions . Similar to the exponential cultures, the ageing bacteria produced alkaline phosphatase, acid phosphatase, leucine arylamidase and naphthol-AS-beta 1-phosphohydrolase, although there was a reduction in the levels of the latter two enzymes . Unlike the other enzymes, urease was not detected in 5-week-old cultures . By using primers based on urease subunit C and 26 kD protein genes for polymerase chain reaction (PCR) these two important gene fragments remained conserved despite the morphological conversion from spiral to coccoid forms . Furthermore, PCR-based random amplified polymorphic DNA (RAPD) fingerprinting showed similar DNA banding patterns from bacteria of various ages, demonstrating the conservation of the DNA composition despite morphological changes . This study shows that the aging coccoid form of H . pylori, although reportedly non-culturable in vitro, remains genetically unchanged indicating that it is likely to be viable. Microbios, 1996, 87(351), 77 - 87 Role of iron in the pathogenesis of Mycobacterium avium infection in mice; Dhople AM et al.; Mycobacterial infections are of serious concern to HIV-infected patients, and take a heavy toll of such patients . Mycobacterium avium is the most common opportunistic bacterial infection in patients with AIDS . The overload of iron in serum has been implicated in the pathogenicity of a number of bacterial infections . Since iron storage in cells such as macrophages is increased in AIDS, the role of iron as a possible factor in the pathogenesis of M . avium infection was examined . Supplementing iron to normal laboratory chow resulted in accelerated M . avium infection in mice inoculated earlier with the same organism . The bacterial loads in liver, spleen and lungs were approximately 12-fold higher in mice receiving iron supplementation compared with control groups . This is attributed to an increased percentage saturation of iron in the sera of the mice, thus making more iron available for the replication of bacteria . The addition of beef fat to the diet, together with high iron supplementation, further enhanced the infection . Using smaller inocula, mice receiving chow supplemented with high iron and fat developed disseminated M . avium infection faster than control mice . The results provide strong evidence that iron may play a major role in the pathogenesis of M . avium infection. Acta Neurochir (Wien), 1996, 138(12), 1431 - 6 A flexible metal ventricular catheter for treatment of complicated and protracted infections of the cerebrospinal fluid spaces: preliminary experiences; Vieweg U et al.; In the management of shunt infection, the use of ventricular catheters made of silicone rubber for the temporary external drainage of cerebrospinal fluid (CSF) is general practice . However, the eradication of the primary source of infection may be hindered by the affinity of bacteria to silicone-based material . Compared to silicone catheters, a metal drainage device for temporary ventriculostomy appears to offer more favourable conditions for successful eradication of the infection . Since metal needles cannot be implanted permanently and since their screw-type fixation precludes attachment to the skulls of infants or small children, we developed a flexible metal catheter . This catheter was used exclusively for the treatment of particularly serious or chronic infections of the CSF spaces . The catheter is made of implantation steel and consists of a corrugated tube that renders it flexible . Cerebrospinal fluid drains into a receptable bulb at the tip of the tube . Tubing of other materials may be connected to the end of the metal catheter for either external or internal drainage . It was implanted as a temporary and later permanent CSF drainage in 7 male patients aged from 4 to 60 years, who suffered from chronic, recurrent ventriculitis (n = 5) with an average of 7 previous surgical revisions, as well as from complex infections (n = 2; basal tuberculous meningitis, brain abscess) . The infections were successfully eliminated in 6 patients . In the remaining patient, the metal catheter for external ventriculostomy had to be removed after 4 days due a leakage of CSF; it was replaced by a silicone catheter and later on by a needle drainage . Other complications, such as secondary infection or intracerebral haemorrhage, did not occur . The average duration of external CSF drainage via the flexible metal catheter was 27 days (range 4-50 days) . In 4 patients, the CSF drainage was converted to a permanent ventriculoperitoneal shunt using a new flexible metal catheter . At the time of post-operative follow-up examination (average = 34 weeks), all shunts were functioning and there was no evidence of infection . In cases of especially complicated and protracted CSF infections, the flexible metal ventricular catheter is a promising device for treatment. Adv Exp Med Biol, 1996, 410, 59 - 64 PO2 affinities, heme proteins, and reactive oxygen intermediates involved in intracellular signal cascades for sensing oxygen; Acker H; The oxygen partial pressure field in different organs ranging from 0 to 100 Torr is likely to be a mirror of oxygen sensitive intracellular signal cascades determining ion channel open probability, metabolic pathway activities and gene expression . High or low PO2 affinities of the particular signal cascade optimize the oxygen sensitive cellular response for adapting organ functions to variations of the oxygen supply conditions . The signal cascades are triggered by an oxygen sensor which is believed to be a heme protein . In some cases reactive oxygen intermediates (ROI) are acting as second messengers revealing these signal cascades as an evolutionary highly conserved principle first described in bacteria. Probl Tuberk, 1996, (6), 25 - 7 {Efficacy of clinical follow-up of patients with tuberculosis of the respiratory organs in terms of their cure under present-day epidemiological conditions}; Kovaleva SI et al.; A total of 884 newly detected patients with tuberculosis of the respiratory organs with degradation were followed up for 9 years (1987-1995) . Control group consisted of 1419 patients with respiratory tuberculosis without degradation . As a result of treatment, the destruction focus was healed in 3 years in 73.4% patients and bacteria no longer released in 77.4% . Remote results (after 3, 6, and 9 years) indicate that adequate treatment helps attain cure during the second and third year of the follow-up . Untimely transfer of the patients into inactive groups (40.4% after 3 years) leads to early relapses of tuberculosis, and thus groups of chronic patients are formed both among tuberculosis patients with initially detected destruction and without it (4.5 and 2.2%, respectively) . The number of clinically cured patients by the end of the ninth year was 56.6 and 77%, respectively; 7.4 and 4.7% patients were still registered in the active groups. Crit Rev Ther Drug Carrier Syst, 1996, 13(3-4), 185 - 223 Current applications of polysaccharides in colon targeting; Hovgaard L et al.; Polysaccharides have over the years been used widely in pharmaceutical, chemical, and biochemical drug delivery . This family of natural polymers has an appeal to the area of drug delivery as it is comprised of polymers with a large number of derivatizable groups, a wide range of molecular weights, varying chemical compositions, and for the most part, a low toxicity and biodegradability, yet a high stability . The main scope of this review is to relate the polysaccharides available now to the rapidly growing field of colonic drug delivery . Polysaccharides have been applied to the area as controlled release coatings, matrices, macromolecular carriers, and biodegradable carriers . Bacterial sources of polysaccharidases as well as a detailed treatise of the enzymatic flora of the colonic region are discussed, followed by a presentation of the polysaccharides available for the purpose of colon-specific drug delivery . A final overview of the various approaches to obtain colon-specific delivery by using polysaccharides and a summary of available in vitro and in vivo testing methods will lead to the conclusion that polysaccharides at this point appear to be very promising compounds for use in obtaining colon-specific drug delivery systems. Boll Chim Farm, 1996 Jan, 135(1), 24 - 30 Synthesis of indole-N-propionitrile, N-propionic acid and N-propionamide derivatives of potential biological activity; El-Desokey SI et al.; Cyanoethylation of a selected group from indole derivatives 1a-i using acrylonitrile in pyridine and potassium hydroxide afforded the corresponding indole-1-propionitrile 2a-h derivatives which readily hydrolysed in basic medium to the corresponding indole-1-propionic acid derivatives 4a-c . A several attempts to cyclize the above products-to the corresponding dihydropyrroloquinolin-4-one derivatives . The indole-1-propionamide derivatives reacted with diethyl carbonate and ethyl acetate under Claisen condensation also, with diazonium salts and nitric acid. Retina, 1996, 16(6), 525 - 9 Sensitivity and predictability of vitreous cytology, biopsy, and membrane filter culture in endophthalmitis; Sharma S et al.; PURPOSE: Forty-seven consecutive patients with endophthalmitis were prospectively studied to: 1) compare the undiluted vitreous biopsy culture and membrane filter culture of vitrectomy cassette fluid; 2) determine the sensitivity and specificity of initial smear examinations in relation to final culture; and 3) characterize vitreous cytology in culture positive and negative endophthalmitis . METHODS: Examinations included smears (Gram stains, Diff Quik {Bacto Laboratories Pvt . Ltd., Liverpool, Australia} and unstained) and aerobic/anaerobic culture of undiluted vitreous biopsy specimens and diluted vitreous collected in the cassette . The cassette fluid was passed through 5-microns polycarbonate filter for cytology and a 0.22-microns polyvinylidene difluoride millipore filter for culture . RESULTS: By either method culture results were positive in 27 (57.4%) patients . Culture positive of the vitreous biopsy alone was 44.6% and cassette fluid alone was 49% (P > 0.05) . In the initial smear examination the sensitivity and specificity of the Gram stain were 66.6% and 84.2%, respectively . comparatively, Diff Quik provided lower sensitivity and specificity at 40.7% and 80.9% respectively . Examination of wet film of vitreous samples did not provide any useful information . Although polymorphs were seen in large numbers in all cultures that yielded positive results on examination, they also were seen in 80% of the sterile samples . There was no significant difference in the quantity of macrophages observed in infected and noninfected samples . CONCLUSIONS: Initial smear examination and cytology have limited roles in the diagnosis of infectious endophthalmitis . Obtaining cultures of both an undiluted vitreous biopsy sample and the vitrectomy cassette fluid has a significant advantage compared with culture of only one sample. Folia Biol (Praha), 1996, 42(5), 261 - 6 Ultrastructure of the tracheal epithelium in rabbits after acetylcholine administration; Konradova V et al.; The ultrastructure of the tracheal epithelium was studied 5 and 20 min after intravenous (i.v.) administration of 0.5 mg of acetylcholine . As a result of this treatment, the goblet cells were overstimulated and damaged, and the mechanism of their secretion was accelerated . Only a mild pathological alteration was encountered in the ciliated cells . According to our classification, the degree of secretory element damage was severe, the injury to the ciliary border was moderate . The morphological signs of the severe impairment of the self-cleaning ability of the epithelium were inspissated mucus and very numerous bacteria in the area among free cilia. Acta Vet Scand, 1996, 37(3), 343 - 50 Recovery rate and quality of embryos from mares inseminated at the first post-partum oestrus; Huhtinen M et al.; The pregnancy rate is lower in mares inseminated at the first post-partum (p.p.) oestrus (40-50%) compared with pregnancy rates in subsequent oestrous cycles (55-65%) . The causes of the lowered pregnancy rate are not fully understood . The aim of the present study was to examine if embryonic defects could be one of the reasons for lowered pregnancy rate . A total of 23 p.p . and 14 non-lactating control mares were flushed 7 days after detection of ovulation . Embryo recovery rate was 48% and 71% in p.p . and control mares, respectively (p = 0.16) . Embryos were photographed, measured, graded and stained with fluorescein diacetate to assess their viability . Thereafter embryos were bisected and stained with Hoechst 33342 to count the cell nuclei . Embryos in both groups were equally viable and the cell numbers were not significantly different . According to morphological evaluation all embryos were classified as excellent or good . Embryos aged 7.3 to 7.6 days (+/-0.25 days) were smaller in the p.p . group than in the control group (p < 0.05) . Forty-seven (9/19) and 8% (1/13) of the uterine swabs, taken before the first insemination, yielded bacteria and neutrophils in p.p . and control mares, respectively . The amount of neutrophils and/or bacteria had no statistically significant effect on embryo recovery rate (p > 0.10) . Recovery of embryos was not related to histological findings in uterine biopsies taken after embryo recovery . Embryo recovery rate in p.p . mares (48%) was similar to previously reported foal heat pregnancy rates (40-50%) . Hence, early embryonic death in utero would not be the most likely reason for lowered pregnancy rate in mares inseminated at the first p.p . oestrus . Sperm transport and oviductal conditions by the time of the first p.p . oestrus would need to be studied to clarify the role of fertilisation failure as the cause of lower pregnancy rate in mares inseminated at foal heat. Chirurgie, 1996, 121(6), 453 - 60 {Genetic control of humoral immune response to xenografts . A monoclonal antibody that causes the hyperacute rejection of cardiac xenografts uses the genes in a native form}; Borie DC et al.; The early phases of the rejection of xenografts exchanged between closely-related species are dominated by a vigorous humoral immune response . We have recently used a linkermediated polymerase chain reaction to generate Ig heavy and light chain specific cDNA libraries to examine the Ig gene control of a prototypic IgM monoclonal antibody, HAR-1, that causes the hyperacute rejection of hamster xenografts . Recombinant clones from the library were screened directly from bacterial colonies by PCR and the nucleic acid sequences of the clones established . Our results demonstrate that the HAR-1 hybridoma is encoded by Ig VH and JH genes in a germline configuration . Comparison of the cDNA sequence for HAR1-VH with the germline equivalent of the gene isolated from newborn LEW liver (provisionally designated VH1.1) showed that the two VH sequences share a nucleic acid identity of 99,3% . Similarly, the HAR-1 monoclonal uses a Ig JH gene that is 98,2% identical with the JH1 nucleic acid sequence available in the GenBank . The use of Ig VH and JH genes in a germline configuration is similar to that seen with polyreactive natural antibodies to infectious agents and autoantibodies . These humoral responses are thought to be the results of the stimulation of a T cell-independent subset of B cells, the B-1a/B-1b subset, that are responsible for producing antibodies that serve as a primitive humoral (natural antibody) defense mechanism against infectious diseases . Our results suggest that the humoral component of the rejection of xenografts in the hamster-to-rat model may represent the stimulation of this type of B cell antibody response by xenogeneic target antigens that share antigenic epitopes with bacteria and other infectious agents. Nephrologie, 1996, 17(6), 333 - 6 {Occurrence of extracapillary glomerulonephritis during Mediterranean boutonneuse fever}; Durrbach A et al.; Boutonneuse spotted fever is a summer and late spring infection caused by Rickettsia Conorii mainly in Middle East countries adjacent to the mediterranean coast (1) . Malignant forms have been described in debilitated patients associated with prerenal azotemia, acute tubular necrosis or renal vasculitis (2, 3) . We report for the first time to our knowledge, the association of an extracapillary glomerulonephritis with a Boutonneuse spotted fever . This observation suggest that Rickettsia Conorii, as other bacteria, could participate to the development of glomerulonephritis. Annu Rev Cell Dev Biol, 1996, 12, 305 - 33 Mitotic chromosome condensation; Koshland D et al.; In this chapter, we review the structure and composition of interphase and mitotic chromosomes . We discuss how these observations support the model that mitotic condensation is a deterministic process leading to the invariant folding of a given chromosome . The structural studies have also placed constraints on the mechanism of condensation and defined several activities needed to mediate condensation . In the context of these activities and structural information, we present our current understanding of the role of cis sites, histones, topoisomerase II, and SMC proteins in condensation . We conclude by using our current knowledge of mitotic condensation to address the differences in chromosome condensation observed from bacteria to humans and to explore the relevance of this process to other processes such as gene expression. Ann Ist Super Sanita, 1996, 32(1), 123 - 31 Genetic instability and methylation tolerance in colon cancer; Aquilina G et al.; Microsatellite instability was first identified in colon cancer and later shown to be due to mutations in genes responsible for correction of DNA mismatches . Several human mismatch correction genes that are homologous to those of yeast and bacteria have been identified and are mutated in families affected by the hereditary non-polyposis colorectal carcinoma (HNPCC) syndrome . Similar alterations have been also found in some sporadic colorectal cancers . The mismatch repair pathway corrects DNA replication errors and repair-defective colorectal carcinoma cell lines exhibit a generalized mutator phenotype . An additional consequence of mismatch repair defects is cellular resistance, or tolerance, to certain DNA damaging agents. Med Tr Prom Ekol, 1996, (8), 16 - 9 {Physiologic and hygienic characteristics of the work of road-building machine operators in hot climate}; Retnev VM et al.; Constructing automobile roads in hot climate area, operators of road construction machines face high neuropsychic loads, heating microclimate, intensive noise and vibration, workplace pollution with dust, carbon and nitrogen oxides, bacteria and fungi . Those factors induce marked stress of central nervous functions, thermoregulatory processes and cardiovascular system of the operators. Adv Exp Med Biol, 1996, 404, 415 - 28 Biological activity of saponins from two Dracaena species; Okunji CO et al.; Many species of the west African "soap tree" Dracaena are used in traditional medicine for the treatment of a variety of diseases . In continuation of our search for anti-infective agents from plants implicated in traditional medicine, we evaluated the biological activities of saponins from extracts of Dracaena mannii and Dracaena arborea by using a battery of test systems such as radiorespirometry, Cytosensor bioautography, and agar dilution methods and molluscicidal tests . Bioassay-directed fractionation of the methanol extracts of seed pulp using a combination of chromatographic techniques, gel filtration, droplet countercurrent chromatography (DCCC), and low-pressure liquid chromatography (Lobar), led to the isolation and characterization of spiroconazole A, a pennogenin triglycoside {3 beta-O-{(alpha-L-rhamnopyranosyl(1-->2), alpha-L-rhamnopyranosyl(1-->3)-beta-D-glucopyranosyl}-17 alpha-hydroxyl-spirost-5-ene} (Fig . 1) . As the active constituent, spiroconazole A exhibited pronounced antileishmanial, antimalarial, and molluscicidal activities . This paper also reports on the fungistatic, fungicidal and bacteriostatic activity of spiroconazole A against 17 species of fungi and 4 of bacteria. Vet Res, 1996, 27(6), 561 - 8 Furunculosis control using a paraimmunization stimulant (Baypamun) in rainbow trout; Ortega C et al.; The effect of Baypamun, a non-specific immune system stimulant, on the treatment and control of furunculosis was studied in experimentally infected rainbow trout . A statistically significant association between the Baypamun treatment and a reduction in cumulative incidence of the clinical symptoms of Furunculosis and the infection level, despite the presence of the bacteria in blood and organs was observed . A reduction in the mortality rate in Baypamun-treated fish was also observed but this was not statistically significant . The results suggest that the treatment of fish diseases with non-specific immune stimulants could be, in the future, a good defense strategy leading to a reduction in disease symptoms and a greater control of the asymptomatic carriers after an outbreak. Crit Rev Immunol, 1996, 16(4), 423 - 40 Structure and function of human IgA Fc receptors (Fc alpha R); Morton HC et al.; Receptors for the Fc region of IgA are expressed by many human cell types, especially phagocytes located in mucosal areas, where IgA is the prevalent antibody isotype . Binding of IgA-opsonized particles (e.g., bacteria, viruses) to Fc alpha R may trigger a plethora of cell-mediated immune effector functions designed to rid the body of the foreign invader . The IgA receptor present on myeloid cells such as neutrophils, eosinophils, and monocytes (Fc alpha RI or CD89) is a transmembrane glycoprotein that binds both IgA isotypes with similar affinity . Genetic characterization showed Fc alpha RI to be a more distantly related member of the Ig receptor gene family . Recently, Fc alpha RI was found to associate with the FcR gamma-chain signaling molecule through a unique charge-based mechanism . Fc alpha RI is, thus, connected to the intracellular machinery via the ITAM signaling motifs located within the cytoplasmic tail of FcR gamma-chain . Evidence exists in support of receptors for IgA (distinct from Fc alpha RI) on human T and B cells . IgA Fc receptors may, therefore, play a role in both the induction and control of an efficient (mucosal) immune response. Rapid Commun Mass Spectrom, 1996, 10(14), 1802 - 6 In situ methylation of nucleic acids using pyrolysis/mass spectrometry; Abbas-Hawks C et al.; Curie-point pyrolysis/triple quadrupole mass spectrometry (Py/MS/MS) has been used with tetramethylammonium hydroxide (TMAH) to conduct in situ methylation of nucleic acid bases . Nitrogen bases in free nucleotides, oligonucleotides, calf thymus DNA and whole bacterial cells reacted in situ during pyrolysis with TMAH to form the methylated bases . Derivatization increased the volatility of the nitrogen bases and the mass of the diagnostic base peaks, thereby removing them from the positions of lower-mass background peaks . The degree of methylation as a function of TMAH concentration for the oligonucleotide, calf thymus DNA, and the whole bacteria samples was determined and found to correlate with the nature of DNA . The methylated bases were identified by their positive-ion electron ionization fragmentation patterns and confirmed with tandem mass spectrometry . The detection of the methylated bases by Py/MS/MS facilitates the goal of identifying the nucleic acids in a complex mixture (i.e . whole bacterial cells) without extraction and prior derivatization. Life Sci, 1996, 59(22), 1891 - 8 Presence of casein immunoreactive epitopes in molluscs, fish and frog; Franchini A et al.; The presence of a group of peptides derived from milk proteins (caseins) was examined by immunocytochemistry in various tissues from invertebrates and lower vertebrates . Phagocytic hemocytes from different species of molluscs, and cells located in the intestine wall or in related glands of invertebrates and lower vertebrates showed immunoreactivity to antibodies to whole casein and related fragments . Several functional tests (cell migration, inhibition test, phagocytosis) using these peptides were performed on the mollusc hemocytes . Only ovine caseinoglycopeptide was able to increase the phagocytic activity of the hemocytes towards bacteria. Pediatr Neurosurg, 1996, 24(5), 275 - 6 A new catheter for use in severe shunt infections and pyocephalus . A technical note; Grant JA et al.; A catheter is described for use in severe shunt infection where the white blood cell count in the CSF exceeds 1,000 cells/mm3 . The catheter allows for simultaneous irrigation and drainage of the ventricular system. IARC Sci Publ, 1996, (139), 189 - 201 Blocking the endogenous formation of N-nitroso compounds and related carcinogens; Bartsch H et al.; Humans are exposed to a wide range of nitrogen-containing compounds and nitrosating agents, such as nitrite, nitrate and nitrogen oxides (NOx), that can react in vivo to form potentially carcinogenic N-nitroso compounds (NOCs), as well as several carcinogenic C-nitro(so) or reactive diazo compounds . Nitrosating agents can also be synthesized endogenously by bacteria and activated macrophages via the nitric oxide (NO) synthase pathway . Endogenous nitrosation can thus occur at many locations in the body, including sites of chronic infection or inflammation . Ascorbic acid, alpha-tocopherol, phenolic compounds, and fruit, vegetable and plant extracts inhibit NOC formation by destroying nitrosating agents . Fresh fruits and vegetables (sources of nitrosation inhibitors) exert a protective effect against various epithelial cancers . In addition to giving rise to nitrosating agents, overproduction of NO in chronic inflammatory conditions leads to the generation of peroxynitrite (a source of oxidative DNA damage and lipid peroxidation) and aldehydes and epoxides derived from lipid peroxidation that yield miscoding exocyclic DNA adducts . Inhibition of the inducible NO synthase and strengthening of the cellular defence system against oxidative stress should block NO-associated DNA damage . This chapter summarizes mechanistic, experimental and human studies that address the inhibition of endogenous formation of NOCs and related carcinogens as a method of chemoprevention. IARC Sci Publ, 1996, (139), 61 - 90 Vegetables, fruit and phytoestrogens as preventive agents; Potter JD et al.; The practice of medicine-both past and present-often involves the prescription of specific foods (almost always plants) or their potent derivatives, to treat a wide spectrum of illnesses . Foods that have been ascribed healing properties include the Cruciferae, the allium family, celery, cucumber, endive, parsley, radish and legumes . Review of the epidemiological data, including both cohort and case-control studies, of all cancer sites strongly suggests that plant foods also have preventive potential and that consumption of the following groups and types of vegetables and fruits is lower in those who subsequently develop cancer: raw and fresh vegetables, leafy green vegetables, Cruciferae, carrots, broccoli, cabbage, lettuce, and raw and fresh fruit (including tomatoes and citrus fruit) . Other data suggest that foods high in phytoestrogens, particularly soy (which contains isoflavones), or high in precursor compounds that can be metabolized by gut bacteria into active agents, particularly some grains and vegetables with woody stems (which contain precursors to lignans), are plausibly associated with a lower risk of sex-hormone-related cancers . The human evidence for these latter associations is not strong . There are many biologically plausible reasons why consumption of plant foods might slow or prevent the appearance of cancer . These include the presence in plant foods of such potentially anticarcinogenic substances as carotenoids, vitamin C, vitamin E, selenium, dietary fibre (and its components), dithiolthiones, isothiocyanates, indoles, phenols, protease inhibitors, allium compounds, plant sterols, and limonene . Phytoestrogens are also derived from some vegetables and berries as well as grains and seeds . Most of the data for the observations on the anticarcinogenic potential of all of these compounds have come from animal and in vitro studies . At almost every one of the stages of the cancer process, identified phytochemicals are known to be able to alter the likelihood of carcinogenesis-occasionally in a way that enhances risk but usually in a favourable direction . For example, glucosinolates and indoles, thiocyanates and isothiocyanates, phenols, and coumarins can induce a multiplicity of phase II (solubilizing and usually inactivating) enzymes; ascorbate and phenols block the formation of carcinogens such as nitrosamines; flavonoids and carotenoids act as antioxidants, essentially disabling the carcinogenic potential of specific compounds; lipid-soluble compounds such as carotenoids and sterols may alter membrane structure or integrity; some sulphur-containing compounds suppress DNA and protein synthesis; carotenoids can suppress DNA synthesis and enhance differentiation; and phytoestrogens compete with estradiol for estrogen receptors in a way that is generally antiproliferative . Consumption of diets low in plant foods results in a reduced intake of a wide variety of those substances that can plausibly lower cancer risk . In the presence of a diet and lifestyle high in potential carcinogens (whether derived from fungal contamination, cooking or tobacco) or high in promoters (such as salt and alcohol), overall risk of cancer at many epithelial sites is elevated . Plant foods appear to exert a general risk-lowering effect; the patterns of exposure to cancer initiators and promoters and of genetic susceptibility may determine the variations in the site-specific risks of cancer seen across populations. Biochimie, 1996, 78(6), 474 - 87 Group II introns: elaborate ribozymes; Jacquier A; Group II introns are found in organelle genomes of plants, fungi and algae as well as in some bacteria . Some group II introns have been shown to self-splice in vitro and thus constitute examples of ribozymes . Their splicing pathway is analogous to the splicing pathway of nuclear pre-mRNA introns . They thus constitute simple models to analyze RNA catalysis of this type of splicing reactions . In this review article, I will summarize our current state of understanding of the ribozyme activity of group II introns and show that their large size correlates with their ability to perform complex tasks . After discussing the similarities found between group II and nuclear pre-mRNA introns, I will briefly evoke how the ribozyme activity of group II introns might be involved in their transposition at the DNA level. Adv Exp Med Biol, 1996, 405, 171 - 82 Chemistry and biological activity of triterpenoid glycosides from Medicago sativa; Timbekova AE et al.; Ten triterpenoid glycosides have been isolated from the roots of Medicago sativa L . (Leguminosae) including eight new glycosides and two previously known, medicagenic acid 3-O-beta-D-glucopyranoside and hederagenin 3-O-{beta-D-glucopyranosyl(1-->2)-alpha-L-arabinopyranoside} . Four new saponins were identified as glycosides of medicagenic acid, four from hederagenin . Their structures were elucidated by a combination of chemical and spectroscopic techniques (IR, EI/MS, NMR 1H and 13C) . In some cases we used fast atom bombardment mass spectra (FAB/MS), NMR 1H difference spectra (homonuclear double resonance, Overhauser effect), NMR 13C partially decoupled spectra (off resonance), and two-dimensional 1H-1H correlating spectra (COSY 1H-1H) . The inhibitory effects of these glycosides on growth and seed germinations and on some phytopathogenic bacteria have also been tested. Electrophoresis, 1996 Jan, 17(1), 12 - 9 Extended theory of the electrophoretic mobility-shift analysis of nonspecific protein-DNA complexes, featuring cooperativity; Coombs RO et al.; The simulated electrophoretic mobility-shift behavior of a model system, in which the nonspecific binding of a protein to a DNA fragment is cooperative, was compared with the experimental behavior of the DNA: histone-like bacterial protein (HU) system . It was concluded that the binding of HU to an 88 bp DNA fragment is, at least, not highly cooperative . The theory of mobility-shift analysis was extended even further to encompass high affinity sequence-specific binding of protein to a DNA fragment followed by weak nonspecific binding, the latter governed by conditional probabilities . In addition to featuring a ladder of incremental protein-DNA complexes, the computed mobility-shift patterns placed emphasis upon stabilization of weak, nonspecific complexes in gel cages. Probl Tuberk, 1996, (1), 40 - 3 {Immunoenzyme analysis for the identification of Mycobacterium bovis by using monoclonal antibodies}; Androsova MV et al.; Three monoclonal antibodies (MAb) were obtained after fusion of mouse BALB/c splenocytes immunised with gamma-irradiated field strains M . bovis and cells of mouse myeloma . Mab specificity was determined at enzyme immunoassay of the bacteria . The antibodies were able to identify various epitopes of the protein (molecular mass under 31 kD) . One of the antibodies obtained served the basis for the test-system intended for rapid identification of M . bovis . The system requires 0.5-1 mg of the bacterial mass. Adv Exp Med Biol, 1996, 398, 285 - 9 Utilization of superoxide anion by indoleamine oxygenase-catalyzed tryptophan and indoleamine oxidation; Hayaishi O; The following is our current working hypothesis concerning the biological significance of IDO induction . When tissues are invaded by virus, bacteria, or parasites, leukocytes and lymphocytes will accumulate at the site and interferon will be produced by these cells in the inflammatory loci . The interferon thus produced is released and interacts with the cell surface to trigger IDO induction in the same or other types of cells . As a consequence of inflammation, superoxide anion is liberated and serves as a substrate for IDO . Although it is possible that some trytophan metabolites may activate the immune system or act as bacteriostatic agents, available evidence does not support this hypothesis . We therefore tentatively conclude that tryptophan is degraded by IDO and depleted, whereby the growth of viruses, bacteria and certain parasites is inhibited, because tryptophan is the least available and therefore most important essential amino acid for their growth. Rev Alerg Mex, 1996 Jan-Feb, 43(1), 13 - 7 {Superantigens and their behavior in the immune response}; Lopez H et al.; Superantigens are a group of molecules produced by bacteria, viruses and parasites that stimulate T lymphocytes in a very potent way . They join the TCR-HLA complex from the outside, not through the classical peptide-bound mechanism, so that they can easily trigger the citokyne production network and produce tissue damage, they can also lead to clonal anergy over the CD4 T cell subpopulations (Th0/Th1/Th2). Scand J Gastroenterol Suppl, 1996, 220, 128 - 31 Ulcer bleeding and perforation: non-steroidal anti-inflammatory drugs or Helicobacter pylori; Svanes C et al.; Non-steroidal anti-inflammatory drug (NSAID)-exposure increases the risk for ulcer perforation by a factor of 5-8 . Recent data from Norway indicate that NSAID exposure is more common in gastric than in prepyloric, pyloric and duodenal perforation . Twenty to 40% of patients with gastric perforation have used NSAIDs; attributable risks are not published . The risk for ulcer bleeding is increased by a factor of 3-5 in NSAID users, with similar effects for stomach and duodenum . NSAID exposure accounts for 20-35% of ulcer bleedings . There is little knowledge about the role of Helicobacter pylori in ulcer complications, a limited importance of the bacteria is indicated in the etiology of both perforation and bleeding . Ulcer complications have a multifactorial origin . NSAIDs account for a limited part of the events . H . pylori infection may play a still undocumented role . Smoking, alcohol and aspirin are other important causal factors. Scand J Gastroenterol Suppl, 1996, 220, 83 - 6 Inflammatory bowel disease: a new assessment; Gitnick G; The etiologies of Crohn's disease and inflammatory bowel disease remain undefined . A growing body of evidence suggests that genetic factors play at least a permissive role while a variety of initiating agents, varying from bacteria to viruses, to a vast array of inert antigens have been postulated, but clear-cut cause and effect relationships have not been established . Recently, a resurgence of interest in Mycobacteria paratuberculosis has developed . Regardless of the initiating cause, growing evidence continues to suggest an immunomodulatory role for the immune system in perpetuating the chronicity of these illnesses . Treatment approaches have been directed against possible initiating agents as well as against the elements which may establish chronicity . Thus currently antibiotics and anti-mycobacterial drugs are being utilized . A variety of anti-inflammatory and/or immunosuppressive agents, including prednisolone, cyclosporine, 6-mercaptopurine, as well as many 5-aminosalicylic acid products comprise a growing armamentarium. J Bone Joint Surg Br, 1996 Jan, 78(1), 92 - 4 Air contamination during skin preparation and draping in joint replacement surgery; Brown AR et al.; Despite the use of ultraclean air, there are still cases of infection in total joint arthroplasty . One possible route by which bacteria may enter the wound is indirectly by contamination of instruments during skin preparation and draping . We found that bacterial air counts were 4.4 times higher during preparation and draping for hip or knee arthroplasty using an unscrubbed, ungowned leg holder than during the operation itself . With the leg holder scrubbed and gowned during preparation and draping, the air counts were reduced but were still 2.4 fold greater than intraoperatively . On some occasions, the air counts during preparation and draping exceeded the standards for ultraclean air irrespective of the attire of the leg holder . We recommend that the leg is held by a scrubbed and gowned member of the team . More importantly, we consider that instrument packs should be opened only after skin preparation and draping have been completed. Scand J Infect Dis, 1996, 28(4), 361 - 6 Evaluation of serological response to Bartonella henselae, Bartonella quintana and Afipia felis antigens in 64 patients with suspected cat-scratch disease; Dupon M et al.; The serological response to Bartonella henselae, B . quintana, and Afipia felis was assessed by an indirect fluorescence antibody test (IFAT) in 64 patients with suspected cat-scratch disease (CSD) recruited from the Bordeaux area in France . Blood samples were collected from 57 patients with chronic lymphadenopathy who underwent lymph-node biopsy with suggestive histopathologic features of CSD, and from an additional 7 patients with suspected CSD who underwent surgical incision and drainage because of lymph-node tenderness . Of the patients, 31 were male and 33 female, with a median age of 27 years (range 2-89) . 69.8% reported cat and/or dog contact . Of the 26/64 (40.6%) patients, serum samples were positive at a titer of 1:100 or more for immunoglobulin G (IgG) antibodies (17 only to B . henselae, 1 only to B . quintana, 3 only to Afipia felis, and 5 to both B . henselae and B . quintana) . IgM or IgA antibodies were also detected in 10 patients with IgG antibodies to B . henselae . 11 (17.2%) of the 64 patient serum samples were positive at a low titer of 1:50 . These data suggested that serological response assessed by standard IFAT is not enough to confirm a CSD diagnosis. Physiol Chem Phys Med NMR, 1996, 28(1), 29 - 33 Viruses--a conundrum; Pandit HM; To understand the pathophysiology of a disease, it is important to know the origin, causes and effects . This also helps to control and to treat the disease . In virus infections, it is difficult and confusing when the origin is sought . In this article it is hypothesized that the viruses, which are nucleoproteins, arise as fragments or broken segments of DNA or RNA . Various factors, such as radiation, toxic chemicals, pollution, are listed as possible causes of such fragmentations . It is logical that these DNA or RNA fragments must come from the genomes or the genes essential for the proliferation or cell division . The symbiotic and parasitic interrelationships of bacteria, plants and animals make the problem more complex and confusing, because all of them thrive and grow in each other cells, thus producing more nucleoproteins of each . Virions contain a small quantum of energy as the initial source of bioenergy to ignite and initiate the complex chemical reactions needed to use the potential energy reservoirs from the host . In this respect viruses can be considered as borderline between the living and the non-living . If one has to develop an effective drug or method for treating virus infections or cancers, the drug must have an antimitotic activity without affecting other normal functions . Such seems to be the case of globin derivatives of sickle cell and thalassemia red blood cells. Hum Mol Genet, 1996, 5 Spec No, 1489 - 94 Mismatch repair defects in human carcinogenesis; Eshleman JR et al.; Mismatch repair defects are carcinogenic . This conclusion comes some 80 years after the original description of a type of familial colorectal cancer in which mismatch repair defects are involved, and from decades of dedicated basic science research into fundamental mechanisms cells use to repair their DNA . Mismatch repair (MMR) was described first in bacteria, later in yeast and finally in higher eukaryotes . In bacteria, one of its roles is the rapid repair of replicative errors thereby providing the genome with a 100-1000-fold level of protection against mutation . It also guards the genome by preventing recombination between non-homologous regions of DNA . The information gained from bacteria suddenly became relevant to human neoplasia in 1993 when the RER phenotype of microsatellite instability was discovered in human cancers and was rapidly shown to be due to defects in mismatch repair . Evidence supporting the role of MMR defects in carcinogenesis comes from a variety of independent sources including: (i) theoretical considerations of the requirement for a mutator phenotype as a step in multistage carcinogenesis; (ii) discovering that MMR defects cause a 'mutator phenotype' destabilizing endogenous expressed genes including those integral to carcinogenesis; (iii) finding MMR defects in the germline of HNPCC kindred members; (iv) finding that such defects behave as classic tumor suppressor genes in both familial and sporadic colorectal cancers; (v) discovering that MMR 'knockout' mice have an increased incidence of tumors; and (vi) discovering that genetic complementation of MMR defective cells stabilizes the MMR deficiency-associated microsatellite instability . Models of carcinogenesis now must integrate the concepts of a MMR defect induced mutator phenotype (Loeb) with the concepts of multistep colon carcinogenesis (Fearon and Vogelstein) and clonal heterogeneity/selection (Nowell). Microbiol Immunol, 1996, 40(1), 27 - 32 A new primer pair for detection of Chlamydia pneumoniae by polymerase chain reaction; Kubota Y; In order to improve the detection and identification of Chlamydia pneumoniae, new primers for polymerase chain reaction (PCR) were designed based on the DNA base sequence within the 53-kDa protein gene, which is specific for C . pneumoniae . The primers permitted the identification of 24 C . pneumoniae strains collected from different geographical locations, but no reaction was observed with C . trachomatis, C . psittaci nor C . pecorum . The primers were unable to amplify the DNA of bacteria commonly related to respiratory tract infections . The positive amplification was achieved with only 9 EBs/assay . Therefore, the new primers seem to be useful in the diagnosis of C . pneumoniae infections. Arch Med Res, 1996 Autumn, 27(3), 319 - 25 Zinc administration prevents wasting in stressed mice; Garcia Tamayo F et al.; Experimentally induced chronic stress can produce severe retardation on the physical development of young animals . Moreover, the chronic stress and its associated secondary malnutrition cause a variable depression on immunity, whose pathogenesis has been related to the excessive production of cytokines and glucocorticoids . When stressful stimuli are excessive, animals increment their anorexia and express a progressively installed wasting syndrome, associated with hypozincemia and susceptibility to infections with high mortality rate . In this work, chronically stressed mice were studied to observe the prophylactic effect of a zinc treatment on the evolution of both their malnutrition and their immune competence . Stress was induced in newborn Balb/c mice by intraperitoneal (IP) injections with heat-killed bacteria for 4 weeks . Following this inductive period, almost all the stressed mice showed a transient wasting syndrome characterized by anorexia, deficient gain of corporal weight, diarrhea, skin infection, reduced antibody response against antigens of red blood sheep cells, and a decreased proliferative response in their Con-A stimulated splenic lymphocytes . However, when the stressed mice received an additional IP treatment with zinc acetate, their clinical condition showed a significant improvement and their immunocompetence was similar to that exhibited by non-stressed mice from the control groups . The results suggest that zinc supplementation can ameliorate the effects of chronic stress on the growth, corporal weight, and immunocompetence of young mice. Eur Urol, 1996, 30(1), 28 - 33 Prostatic abscesses in sub-Saharan Africa: a hospital-based experience from Cameroon; Angwafo FF 3rd et al.; Abscesses of the prostate are encountered less frequently today and the bacteria responsible have changed over the years . Herein we report our experience with 6 patients with varying presentations . The role of imaging techniques in diagnosis and percutaneous needle aspiration of prostatic abscesses is highlighted. Chirurg, 1996 Jan, 67(1), 1 - 6 {Ulcer--an infection?}; Eissele R et al.; In the past gastric acid was considered to be the major factor in the pathogenesis of peptic ulcers . For the first time bacteria were found in the stomach at the end of the last century . However, Helicobacter pylori could be detected and characterized not before 1983 . Specific factors of virulence enable H . pylori to colonize the gastric mucosa . Infection occurs during childhood and due to a cohort-phenomenon the prevalence increases continuously with age . H . pylori infection can be diagnosed in about 95% of patients with duodenal ulcer and 70% of gastric ulcers . The important pathogenetic role of H . pylori infection could be demonstrated in eradication trials . According to our present knowledge gastric acid and H . pylori are independent prerequisits in peptic ulcer pathogenesis . H . pylori negative ulcer derive from NSAIDs or from the rare Zollinger-Ellison syndrome. Fundam Appl Toxicol, 1996 Jan, 29(1), 40 - 7 Effect of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on influenza virus host resistance in mice; Burleson GR et al.; 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) causes numerous immunotoxic effects including thymic involution and an immunosuppression of nonspecific as well as specific cell- and humoral-mediated immunity . TCDD administration to laboratory animals also results in a decreased resistance to numerous bacteria, viruses, and parasites . Effects on virus host resistance appear to be among the most sensitive effects of TCDD immunotoxicity . However, previous studies have not achieved a no effect level . The present studies utilized an influenza virus host resistance model in mice to quantify the sensitivity of this model to TCDD and to determine the NOAEL (no observed adverse effect level) of TCDD for influenza virus . Results indicated that a single dose of TCDD at 0.10, 0.05, or 0.01 microgram/kg resulted in an increased mortality to Hong Kong influenza virus when mice were challenged 7 days after TCDD administration . Increased mortality was not correlated with increased virus titers in the lungs . TCDD at 0.005 or 0.001 micrograms/kg had no effect on influenza-induced mortality . TCDD alone did not affect thymus weight at any dose administered in this study . TCDD also did not alter the virus-enhanced increase in lung weight:body weight ratio nor the virus-induced decrease in thymus weight . Thus, low levels of TCDD exposure lead to enhanced mortality to influenza virus; however, the mechanism of this effect remains to be elucidated . Nonetheless, enhanced mortality to influenza virus in mice following a single dose of 10 ng TCDD/kg represents the most sensitive adverse effect yet reported for TCDD. J Heart Valve Dis, 1996 Jan, 5(1), 58 - 65 In vitro endothelialization of bioprosthetic heart valves; Fischlein T et al.; BACKGROUND AND AIMS OF THE STUDY: The main disadvantage of xenograft bioprosthetic heart valves is limited durability due to tissue degeneration . METHODS: To study possible alternatives to conventional bioprosthetic heart valves, glutaraldehyde preserved porcine aortic valve leaflets were precoated with fibronectin-heparin and acidic fibroblast growth factor (aFGF) to enhance cell proliferation . Furthermore, different methods of storage and preservation (1.0% benzoic acid, 1.0% sorbic acid, 0.05% and 0.5% dialdehyde starch) were compared to conventional preservation procedures . Valve leaflets were lined with adult human saphenous vein endothelial cells (1.0 x 10(4) AHSVEC/cm2) . The different methods of preservation were compared to leaflets precoated additionally with 1.0 microgram/cm2 aFGF protein, derived from gene manipulated bacteria . Endothelial growth kinetics were studied and verified by an increase of activation following 3H-thymidine incorporation, while the maintained metabolic cell activity was demonstrated by prostacyclin (PGI2) release measurements . In subsequent experiments in vivo degeneration and mineralization were determined by subcutaneous implantation in rats for up to 40 days . RESULTS: Our results favored alternatively treated valve leaflets; endothelial cells grew to persistent monolayers in six to 12 days in contrast to conventional processed leaflets where no endothelial cell growth was possible . Atomic absorption spectroscopic assessment of subdermal valve implants showed significantly lower contents of calcium, magnesium and phosphate in alternatively treated leaflets . Furthermore, precoating of alternatively preserved valve leaflets with aFGF protein and endothelial cells improved in vitro and in vivo results significantly . CONCLUSIONS: Our study shows that endothelial cell growth as well as significantly reduced in vivo degeneration and mineralization of valve leaflets may be feasible if bioprosthetic heart valves are processed according to alternative, non-toxic conservation procedures and are precoated with angiogenic growth factor protein. Int Orthop, 1996, 20(3), 142 - 6 Preparation of bank bone using defatting, freeze-drying and sterilisation with ethylene oxide gas . Part 1 . Experimental evaluation of its efficacy and safety; Kakiuchi M et al.; We devised a method of sterilising bone allografts which consists of defatting in chloroform and methanol, freeze-drying and sterilisation with ethylene oxide gas . The purpose of defatting and freeze-drying was to facilitate subsequent sterilisation by eliminating the barrier to diffusion of the gas into bone, to lower residual levels of ethylene oxide and its toxic by-products, to eliminate alloantigens and to make storage possible at room temperature . The efficacy and safety of the method were evaluated by testing the sterilisation of infected bone from 6 patients with active chronic osteomyelitis, the penetration of ethylene oxide into human femoral heads treated by this or by freeze-drying or freeze-thawing, and the desorption of ethylene oxide and its toxic by-products from pieces of bone treated by these methods . All the samples of infected bone tested negative for bacteria after treatment . The gas penetrated into the central area of the femoral heads in a few hours . Residual levels of ethylene oxide and its toxic by-products were much lower in the treated bone than in freeze-dried or freeze-thawed bone, and decreased quickly in flowing air . Prior defatting and freeze-drying facilitated penetration of ethylene oxide into bone during sterilisation and the desorption of ethylene oxide and its toxic by-products after sterilisation . Preparation under clean, but not sterile, conditions and storage at room temperature make bone banking more practical and efficient. J Pak Med Assoc, 1996 Jan, 46(1), 5 - 7 Study the presence of fibronectin binding protein (FnBp) in tuberculosis (TB) patients by elisa; Ahmad A et al.; Fibronectin-binding protein (FnBp) antigens are a prominent secretory protein of short term culture supernatants of M . tuberculosis and M . bovis (BCG) and is conserved within the genus Mycobacterium . The 30/31 kDa antigen of M . tuberculosis is one of the major secretory molecules and is probably routinely recognised by the host immune system in the early stage of tuberculosis infection . Serum immune complexes, prepared from TB patients and normals, were analysed for the presence of FnBp by ELISA using an anti-30/31 kDa (FnBp) monoclonal antibody (CF8) and by western blotting using Fibronectin-HRP . A significant difference was seen between normals and TB patients (p < 0.05) . This test was found to have a specificity of 80% and a positive predictive value of 73% . This is a preliminary finding and the test needs to be evaluated further for its performance on a larger number of confirmed TB patients and controls. Acta Vet Hung, 1996, 44(1), 1 - 8 Bovine leukocyte adhesion deficiency: a brief overview of a modern disease and its implications; Gerardi AS; Bovine Leukocyte Adhesion Deficiency (BLAD) is a genetic disease of cattle affecting the hematopoietic system . In the last decade BLAD has become a disease of economic importance in the dairy industry . As such, this overview describes the chronological developments and thinking that led to the elucidation of BLAD as a distinct disease entity from previous models in canine and human populations . All species affected exhibit symptoms of chronic and recurrent infections . Necrotic and/or gangrenous infections of soft tissues are prevalent, as well as secondary infections with bacteria or fungi . Low birthweight and unthriftiness are key symptoms of neonates in all species affected by LAD . Dermatomycoses and impaired pus formation are also common findings . The physiological basis for BLAD is a deficiency in leukocyte (particularly neutrophil) chemotactic and phagocytic properties . The inhibition of diapedesis in the inflammatory response prevents normal immune reactions to invading pathogens . Chronic infections are a consequence of the faulty immune mechanisms . The biochemical etiology of BLAD involves cell surface glycoprotein molecules known as integrins . These are responsible for cell-cell interactions necessary for neutrophils to adhere to vascular endothelium in a normal individual . Experiments using monoclonal antibodies to block LFA-1, Mac-1, and p150,95 (three integrins vital for cell-cell interactions) mimic BLAD symptomatology and have led to the discovery of the reciprocal Intercellular Adhesion Molecule (ICAM) . Through pedigree analysis and biochemical detection with restrictive endonucleases BLAD has been isolated genetically to a single gene locus . The economic significance and prophylaxis are briefly discussed . In addition, the beneficial aspects of the study of BLAD are addressed . There are advantages of producing a BLAD-like state in preventing transplant rejection, ischemia-reperfusion injury, and other scenarios arising from the deleterious effects of the inflammatory response. Mol Microbiol, 1996 Jan, 19(2), 397 - 405 The TraB protein, which mediates the intermycelial transfer of the Streptomyces plasmid pSN22, has functional NTP-binding motifs and is localized to the cytoplasmic membrane; Kosono S et al.; The traB gene on the Streptomyces conjugative plasmid pSN22 is required for intermycelial plasmid transfer and the mobilization of chromosomal markers (Cma) . The predicted amino acid sequence of TraB contains one Walker type-A and two type-B NTP-binding motifs . Site-directed mutagenesis revealed that the type-A motif and one of the type-B motifs, 109 amino acid residues downstream of the type-A motif, were essential for both plasmid transfer and Cma . The second type-B sequence could be changed without any phenotypic effect . A modified traB gene was constructed, resulting in the production of a functional protein with an amino-terminal c-Myc epitope tag for immunological analysis . This protein was associated with the cytoplasmic membrane, suggesting that TraB is a membrane protein that uses energy from ATP hydrolysis to transport DNA between mycelia . The c-Myc tagging of TraB decreased the efficiency of intramycelial plasmid spread, suggesting that TraB is involved in both inter- and intramycelial transfer processes. Clin Infect Dis, 1996 Jan, 22(1), 59 - 62 Effect of inactivated poliovirus vaccine on the antibody response to Bordetella pertussis antigens when combined with diphtheria-pertussis-tetanus vaccine; Halperin SA et al.; To determine if inactivated poliovirus vaccine combined with diphtheria and tetanus toxoids and whole-cell pertussis vaccine interferes with the immunogenicity of pertussis vaccine, we performed a randomized trial of diphtheria and tetanus toxoids and pertussis vaccine combined with inactivated poliovirus vaccine given as a single injection or as two separate injections at the same visit to infants immunized at 2, 4, and 6 months of age . A total of 84 infants were enrolled in the study; 44 received the single injection, and 40 received separate injections . Before immunization, there were no differences in antibody values between the two groups . After two vaccine doses, infants immunized with the single injection had significantly lower values of antibody to filamentous hemagglutinin (8.3 vs . 23.7 ELISA units; P < .001), fimbriae (266.5 vs . 771.8 ELISA units; P < 0.01), and the 69-kD membrane protein (442.2 vs . 1,352 ELISA units; P < .001) . After the third dose, these differences persisted, and differences were also detected for antibody to pertussis toxin (10.0 vs . 35.5 ELISA units; P < .001) and a whole-bacteria antigen preparation (2,667 vs . 3,829 ELISA units; P < .05) . We conclude that there is a diminished antibody response to the pertussis vaccine when inactivated poliovirus vaccine is combined with the diphtheria and tetanus toxoids and pertussis vaccine. Mol Microbiol, 1996 Jan, 19(1), 187 - 95 Purification of the Tn10-specified tetracycline efflux antiporter TetA in a native state as a polyhistidine fusion protein; Aldema ML et al.; The bacterial tetracycline-resistance determinant from Tn10 encodes a 43 kDa membrane protein, TetA, responsible for active efflux of tetracyclines . The tetA gene was cloned behind a T7 promoter/lac operator in a plasmid that provided fusion of TetA to a polyhistidine-carboxy terminal tail . A second plasmid provided a regulated T7 RNA polymerase . The specific activity of the TetA fusion protein was between 10-40% that of the wild-type protein as assayed by tetracycline resistance in cells and by transport in membrane vesicles . The fusion protein, overproduced approximately 3-13-fold, was purified by nickel chelation chromatography . Calculations from circular dichroism spectra of the purified protein solubilized in dodecylmaltoside gave an alpha-helix content of 54-64%, close to the 68% predicted from the amino acid sequence by hydropathy analysis (12 membrane-spanning helices) for the native protein in the membrane bilayer . Fluorescence studies showed binding activity of the purified protein to its substrate, the tetracycline analogue 13-(cyclopentylthio)-5-hydroxy-6-alpha-deoxytetracycline . These findings suggested that the purified protein was in a native state. Shock, 1996 Jan, 5(1), 72 - 5 Hepatic reticuloendothelial system dysfunction after intestinal ischemia-reperfusion; Haglind E et al.; The functional capacity of the hepatic reticuloendothelial system (RES) was studied under conditions of intestinal shock . Wistar rats underwent intestinal ischemia (60 min)-reperfusion or a sham procedure . One-half of each group received intravenous infusion of 2.6 x 10(-6) g/kg lipopolysaccharide B . Phagocytic clearance and phagocytic killing by the hepatic RES were quantitated using a double label bacteria clearance assay . Hepatic phagocytosis was unchanged 3 h after reperfusion of the ischemic intestine but hepatic killing was significantly decreased compared with the sham situation . The results were not different in animals who had received LPS B infusion . The plasma endotoxin levels correlated inversely to the hepatic killing efficiency (r = -.801) in the shocked animals given exogenous endotoxin . The decreased capacity of the hepatic RES to degrade bacteria present in the portal circulation could be an important pathophysiologic mechanism in the development of bacterial translocation. Microsc Res Tech, 1996 Jan 1, 33(1), 2 - 11 Alveus, gen . nov . (Bacillariaceae, Bacillariophyta), a heavily silicified diatom found in warm water oceans; Kaczmarska I et al.; Scanning electron microscope images of complete and damaged frustules of Nitzschia marina Grun . revealed an unusual chambered structure of the valve . This structure is not present in any existing nitzschioid genus and warrants an independent generic status for this taxon, which is therefore separated into Alveus gen . nov . Generic and emended specific definitions are provided and presented in the context of their relationship to other genera within the family Bacillariaceae . Deep relief of the inner surface of Alveus valves is similar to that of genera Fragilariopsis, Denticulopsis, Neodenticula, and Crucidenticula . It differs from all these diatoms in the alveolar striation . This heavily silicified diatom was found in samples from the US Joint Global Oceanographic Flux Studies cruise TT007 stations 2 and 4, (9 degrees and 5 degrees N respectively, 140 degrees W) in the equatorial Pacific . Presently, this uncommon species is restricted to warm waters . It is very common in the surface sediments of the equatorial Pacific and Indian Ocean and in the tropical and subtropical Atlantic . The sedimentary record of Alveus marinus goes back to the Middle Late Miocene (approximately 8 Ma years). J Clin Lab Anal, 1996, 10(4), 193 - 204 Epitope mapping of a recombinant human TSH receptor extracellular domain: identification of a predominant epitope using animal sera; Hunt N et al.; The extracellular domain of the TSH receptor (TSHR-561, amino acids #78-389) was expressed as a hexa-histidine fusion protein in bacteria . The recombinant protein was purified to homogeneity and used to immunize porcine and ovine species . High titre antibodies were obtained from both species that recognized the recombinant protein in Western blot analysis but failed to interfere with the TSH radio receptor assay . An epitope library was constructed and screened with affinity purified ovine and porcine antisera and detected a number of positive clones . Sequence analysis revealed that all of the epitopes contained sequences derived from the carboxyl terminus of the recombinant immunogen . One clone defined an epitope covering 16 amino acids from the carboxyl terminus and was the common epitope found in all of the other clones . Western blot screening of a large panel of Graves' sera with recombinant TSH receptor protein identified one patient sera that also recognized linear epitopes in the TSHR-561 protein . Experimentation demonstrated that the linear epitope recognized by this human sera was identical to the sequence recognised by the animal antisera . This sequence is unique to the TSH receptor and will be useful in further studies to analyze the TSH receptor protein. Annu Rev Biochem, 1996, 65, 441 - 73 Structural basis of lectin-carbohydrate recognition; Weis WI et al.; Lectins are responsible for cell surface sugar recognition in bacteria, animals, and plants . Examples include bacterial toxins; animal receptors that mediate cell-cell interactions, uptake of glycoconjugates, and pathogen neutralization; and plant toxins and mitogens . The structural basis for selective sugar recognition by members of all of these groups has been investigated by x-ray crystallography . Mechanisms for sugar recognition have evolved independently in diverse protein structural frameworks, but share some key features . Relatively low affinity binding sites for monosaccharides are formed at shallow indentations on protein surfaces . Selectivity is achieved through a combination of hydrogen bonding to the sugar hydroxyl groups with van der Waals packing, often including packing of a hydrophobic sugar face against aromatic amino acid side chains . Higher selectivity of binding is achieved by extending binding sites through additional direct and water-mediated contacts between oligosaccharides and the protein surface . Dramatically increased affinity for oligosaccharides results from clustering of simple binding sites in oligomers of the lectin polypeptides . The geometry of such oligomers helps to establish the ability of the lectins to distinguish surface arrays of polysaccharides in some instances and to crosslink glycoconjugates in others. Am J Ind Med, 1996 Jan, 29(1), 3 - 13 Endotoxin exposure-response in a fiberglass manufacturing facility; Milton DK et al.; Peak expiratory flow (PEF) and workplace exposure to endotoxin, phenolic resin, and formaldehyde were measured to investigate asthma symptoms and medication use among employees in a fiberglass wool manufacturing plant . Self-recorded PEF was obtained from 37 workers, for a total of 181 days off work and 187 days at work with concurrent personal exposure monitoring . Pre- and post-shift spirometry were obtained on at least 2 days . The 8 hr time-weighted average personal exposure ranges were endotoxin; 0.4-759 ng/m3; phenolic resin, 5.7-327 micrograms/m3; and formaldehyde, 1.2-265 micrograms/m3 . Amplitude percent mean peak flow was associated with years since starting regular work in the highest endotoxin exposure area, although current assignment in that area was associated with reduced amplitude--evidence for a healthy worker effect . Exposure-response was analyzed by regression of lung function change on exposure using generalized estimating equations with robust variance estimates . Endotoxin exposure above 4 ng/m3 (8 hr time-weighted average) was associated with a decline in lung function across the work shift, and with drops in lung function 16-20 hr after exposure . Phenolic resin exposure was not consistently associated with decrements, and formaldehyde was not associated with decrements in lung function. Arch Virol Suppl, 1996, 11, 171 - 9 Relative importance of the enterically transmitted human hepatitis viruses type A and E as a cause of foreign travel associated hepatitis; Langer BC et al.; Hepatitis contracted during a stay abroad may be caused by a wide range of pathogens including viruses, bacteria, protozoa or helminths . In many cases, the etiological agent primarily infects other target organs and tissues, involving the liver either as part of a disseminated infection or secondarily to mechanical biliary tract obstruction . The article focuses on enterically transmitted hepatitis caused by the primarily hepatotropic human hepatitis viruses type A and E and discusses their importance in travel-related disease. Int J Legal Med, 1996, 108(6), 323 - 6 The detection of picoplankton 16S rDNA in cases of drowning; Kane M et al.; Picoplankton belonging to the Synechococcus genus in cyanobacteria (approximately 1 micron in size) are found ubiquitously in Lake Biwa, Japan . However, they could not be morphologically discriminated from other bacteria by microscopy . In this study we attempted to use picoplankton for the diagnosis of drowning by PCR analysis of the 16S ribosomal DNA (rDNA) . We designed primers complementary to the variable regions of 16S rDNA of the picoplankton we had sequenced . A comparison was made of the PCR products from the three picoplanktons, five other cyanobacteria, Melosira (diatom), Staurdstrum (green alga), bacteria from Lake Baikal, and humans . The picogram order of template DNA from picoplankton was specifically amplified by the primers . When the template of picoplankton was mixed with human lung tissue, at least 10 ng of template DNA was needed to obtain a PCR product . The isolation of the picoplankton from human lung tissue increased the sensitivity of PCR more than a hundred-fold . The specific PCR products of the picoplankton were obtained from formalin-fixed drowning tissue . Molecular biological diagnosis of drowning was successful using picoplankton 16S rDNA. Am J Reprod Immunol, 1996 Jan, 35(1), 43 - 50 Identification of the complement regulatory protein CD59 in human colostrum and milk; Bjorge L et al.; PROBLEM: Complement lytic activity has been demonstrated, and a potential for its activation is present in human colostrum and milk . This necessitates the presence of regulatory mechanisms protecting epithelial cells in the oropharynx and the gastrointestinal tract of the infant, the milk cellular elements, and bacteria colonizing the oropharynx and the gastrointestinal tract . Lactoferrin and C1 inhibitor have been attributed such a role . However, it is likely that additional protection against the cytolytic activity of the membrane attack complex is required . This has lead us to investigate the presence of the complement regulatory protein CD59 in human colostrum and milk, and to further characterize the source of secretion . METHOD: Samples of human colostrum and milk were obtained from volunteers at different stages of lactation, and separated into fat, skim milk, and milk cellular elements by centrifugation . Normal human mammary gland tissues were obtained from patients undergoing biopsy for benign conditions . SDS-PAGE and Western blotting, and an immuno dot-blot assay were used to identify CD59 in human milk . Immunohistochemistry was performed on all tissue samples and cytospins of the milk cellular elements, using monoclonal antibodies to CD59 . RESULTS: CD59 was present in cell-free colostrum and milk as a 19-25 kDa glycoprotein . No variation in CD59 levels was detected between colostrum and milk . CD59 was present in great amounts in the cytoplasm and was highly expressed on the surface membrane on mammary gland acinar and ductal epithelial cells, while the milk cellular elements contained CD59 mainly in their cytoplasm . CONCLUSION: The complement regulatory protein CD59 present in cell-free human colostrum and milk may exert its effects both in the mammary gland and in the oropharynx and gastrointestinal tract of the infant . The lobuloalveolar epithelial cells in the mammary gland are the likely source of secretion. Am J Physiol, 1996 Jan, 270(1 Pt 1), C12 - 30 Water transport across mammalian cell membranes; Verkman AS et al.; This review summarizes recent progress in water-transporting mechanisms across cell membranes . Modern biophysical concepts of water transport and new measurement strategies are evaluated . A family of water-transporting proteins (water channels, aquaporins) has been identified, consisting of small hydrophobic proteins expressed widely in epithelial and nonepithelial tissues . The functional properties, genetics, and cellular distributions of these proteins are summarized . The majority of molecular-level information about water-transporting mechanisms comes from studies on CHIP28, a 28-kDa glycoprotein that forms tetramers in membranes; each monomer contains six putative helical domains surrounding a central aqueous pathway and functions independently as a water-selective channel . Only mutations in the vasopressin-sensitive water channel have been shown to cause human disease (non-X-linked congenital nephrogenic diabetes insipidus); the physiological significance of other water channels remains unproven . One mercurial-insensitive water channel has been identified, which has the unique feature of multiple overlapping transcriptional units . Systems for expression of water channel proteins are described, including Xenopus oocytes, mammalian and insect cells, and bacteria . Further work should be directed at elucidation of the role of water channels in normal physiology and disease, molecular analysis of regulatory mechanisms, and water channel structure determination at atomic resolution. Genetics, 1996 Jan, 142(1), 313 - 26 Bayesian procedures for the estimation of mutation rates from fluctuation experiments; Asteris G et al.; Bayesian procedures are developed for estimating mutation rates from fluctuation experiments . Three Bayesian point estimators are compared with four traditional ones using the results of 10,000 simulated experiments . The Bayesian estimators were found to be at least as efficient as the best of the previously known estimators . The best Bayesian estimator is one that uses (1/m2) as the prior probability density function and a quadratic loss function . The advantage of using these estimators is most pronounced when the number of fluctuation test tubes is small . Bayesian estimation allows the incorporation of prior knowledge about the estimated parameter, in which case the resulting estimators are the most efficient . It enables the straightforward construction of confidence intervals for the estimated parameter . The increase of efficiency with prior information and the narrowing of the confidence intervals with additional experimental results are investigated . The results of the simulations show that any potential inaccuracy of estimation arising from lumping together all cultures with more than n mutants (the jackpots) almost disappears at n = 70 (provided that the number of mutations in a culture is low) . These methods are applied to a set of experimental data to illustrate their use. Clin Diagn Lab Immunol, 1996 Jan, 3(1), 73 - 8 A sensitive and specific PCR method to detect Helicobacter felis in a conventional mouse model; Kong L et al.; Although many detection methods have been used to determine Helicobacter colonization in small animal models, the sensitivity and specificity of these detection methods are limited . To improve the Helicobacter felis conventional mouse model for accurate evaluation of therapeutic regimens, we developed a PCR for detection of, and a competitive PCR for quantitation of, H . felis in viral antibody-free (VAF) mice . The PCR was based on the H . felis 16S rRNA gene . An internal control DNA was used for competitive quantitation of the PCR . VAF conventional Swiss-Webster mice were infected with an H . felis culture by oral gavage . At various times after H . felis challenge and therapy, stomach mucosa was collected and evaluated by PCR . PCR detected approximately 50 to 100 H . felis cells per mouse stomach and showed no cross-reaction with other bacteria commonly found in mouse stomachs . Colonization of H . felis in the mouse stomach was confirmed by culture isolation from germfree mice and histological examination of VAF mice . Response to therapy in this H . felis model correlated well with results seen in human clinical trials with H . pylori . A model utilizing PCR detection which may be useful for discovering new antibiotics and/or vaccines against Helicobacter ulcer disease has been developed. Clin Diagn Lab Immunol, 1996 Jan, 3(1), 66 - 72 PCR detection of colonization by Helicobacter pylori in conventional, euthymic mice based on the 16S ribosomal gene sequence; Smith JG et al.; Many animal models of Helicobacter infection have been described, including infection in rhesus monkeys, ferrets, gnotobiotic piglets, and mice . These animal models utilize a combination of detection methods, including culture, urease testing, and histopathology, all of which may be unreliable, insensitive, or labor-intensive . Development of new animal models of Helicobacter pylori requires new methods of detection with increased sensitivity and specificity . We have developed sensitive and specific PCR primers based on the 16S ribosomal gene sequence of H . pylori . The primers detected single-copy 16S DNA representing 0.2 cell of pure H . pylori (2 cells in the presence of mouse stomach mucosal DNA) and did not cross-react with closely related bacteria . We were able to detect colonization by H . pylori in conventional, euthymic, outbred mice up to 4 weeks postinoculation with a high percentage of isolates tested . One isolate of H . pylori was detected by PCR in 100% of the mice at 6 months and 60% of the mice 1 year after inoculation . Approximately 10(3) to 10(4) H . pylori cells per stomach were detected by utilizing this PCR methodology semiquantitatively . These primers and PCR methodology have facilitated detection of H . pylori colonization in conventional, euthymic mice, colonization which may not have been detectable by other methods. Clin Diagn Lab Immunol, 1996 Jan, 3(1), 47 - 50 Differences between Taylorella equigenitalis strains in their invasion of and replication in cultured cells; Bleumink-Pluym NM et al.; The ability of Taylorella equigenitalis, the causative agent of contagious equine metritis, to invade and replicate in equine derm cells was studied . The kinetics of invasion and replication were determined for four T . equigenitalis strains . On the basis of these experiments, a simpler assay in which the invasive as well as the replicative properties of a particular strain could be determined was developed . This assay was used to characterize 32 strains, which had previously been typed by field inversion gel electrophoresis of genomic restriction fragments . The invasiveness of T . equigenitalis strains ranged from 3 to 0.015 bacteria per cell and seemed to be associated with the contagiousness of the infection . The replication index (number of intracellular bacteria per cell at 24 h after inoculation divided by the number of intracellular bacteria per cell at 4 h after inoculation) varied from 1 to 857 and seemed to be associated with the severity of the symptoms of contagious equine metritis . There was no association between the invasiveness and the replication index of the strains, nor was there an association of invasion and replication with field inversion gel electrophoresis grouping. Cell Motil Cytoskeleton, 1996, 34(2), 108 - 21 Partial characterization of zeugmatin indicates that it is part of the Z-band region of titin; Turnacioglu KK et al.; Zeugmatin is a muscle specific protein discovered by Maher et al . {1985: J . Cell Biol . 101:1871-1883} to be in Z-Bands of muscle and in the dense bodies of smooth muscle . Maher et al . {1985} generated a zeugmatin specific monoclonal antibody, McAb20, and then used immunoaffinity chromatography to isolate a 600-800 kD protein . During myofibrillogenesis of embryonic cardiac muscle, zeugmatin is detected in fully formed Z-bands in the mature myofibrils but not in the Z-bodies of premyofibrils {Rhee et al., 1994: Cell Motil . Cytoskeleton 28:1-24} . Rhee et al . {1994} have postulated that zeugmatin may be responsible for the fusion of the alpha-actinin containing Z-bodies to form the solid Z-Bands of the mature myofibrils . The current studies were undertaken to characterize the properties of zeugmatin . The McAb20 was used to probe a chicken heart lamba gt11 expression library, and three unique positive clones of 1.1, 1.4, and 1.7 kB were isolated . These were inserted into pcDNA3, sequenced, and assembled into a 1.8 kB ORF . A 60% identity with N-terminal region of the human cardiac titin sequence was revealed at the amino acid level . This region of the 1.8 kB zeugmatin sequence is located entirely in the Z-band region of the human cardiac titin molecule . The 1.1 kB clone of zeugmatin was subcloned into pTrcHisC and expressed in bacteria . Bacterial lysates were prepared and run over nickel columns to isolate a 46 kD fusion protein . This fusion protein formed a complex with purified alpha-actinin that could be immunoprecipitated with the zeugmatin specific antibody, McAb 20 . The 1.1 kB sequence was transfected into non-muscle cell lines, PtK2 and REF . Twenty-four hours after transfection, the 46 kD zeugmatin peptide, not present in control non-muscle cells, was localized in focal adhesions and in a punctate pattern along the stress fibers . Double immunofluorescence staining revealed that zeugmatin colocalized with the alpha-actinin in the dense bodies and focal contacts of the stress fibers . At longer time points, as the transfected cells accumulated more truncated zeugmatin molecules, the cells lost adhesion plaques and stress fibers, and became detached from the substratum . Our results indicate the zeugmatin is part of the titin molecule that is located within the Z-band and that this section of the titin molecule anchors the actin crosslinking alpha-actinin molecules. Verh K Acad Geneeskd Belg, 1996, 58(2), 93 - 116 {Significance of intestinal inflammation in the pathogenesis of spondylarthropathies}; Mielants H et al.; The concept of spondylarthropathy (SpA) gathers together a group of chronic diseases with common clinical, biological, genetic and therapeutic characteristics . The concept forms a distinct entity, different from other rheumatic diseases . The target organs are not only the joint, but also the axial skeleton, the enthesis, the eye, the gut, the urogenital tract, the skin and sometimes the heart . The prevalence of this entity in the general population is estimated 1%, equal to the prevalence of rheumatoid arthritis . Genetical predisposition (HLA-B27) is one of the clues to the pathogenesis of the disease . Since reactive arthritis is induced by specific urogenital or enterogenic bacteria, and since the gut is implied in different forms of spondylarthropathies, especially in IBD, it was clear that the gut could play an important role by permitting exogenous factors to enter the body . This hypothesis was the rationale for investigating the gut in the spondylarthropathies by performing ileo-colonoscopies . In the first ileo-colonoscopic studies of SpA patients, histological signs of gut inflammation were found in a relatively great number of patients, mostly not presenting any clinical intestinal manifestations . These lesions were not seen in other inflammatory joint diseases . Further ileo-colonoscopic studies confirmed the strong relationship between gut and joint inflammation . In patients in whom a second ileo-colonoscopy was performed, remission of the joint inflammation was always connected with a disappearance of the gut inflammation, whereas persistence of locomotor inflammation was mostly associated to the persistence of gut inflammation . The hypothesis was proposed that some patients with a spondylarthropathy had a form of subclinical Crohn's disease in which the locomotor inflammation was the only clinical expression . This hypothesis was confirmed in prospective long-term studies in which the ileo-colonoscopied patients were reviewed 2 to 9 years later:about 6% of SpA patients not presenting any sign of Crohn's disease at first investigation but demonstrating gut inflammation on biopsy, developed full-blown Crohn's disease . By performing electronmicroscopy it was described that in patients with SpA the number of membranous (M) cells, which are scarce in normal ileum, is increased in number in inflamed mucosa . They showed a thin rim of cytoplasm covering groups of lymphocytes . In chronic inflammatory lesions necrotic M-cells, rupture of M-cells and lymphocytes entering the gut lumen was observed . The bursting of M-cells at the top of the lymphoid follicles leads to interruption of the gut epithelial lining and gives the luminal content access to the lymphoid tissue . This can be responsible for an exponential increase of local antigen stimulation . Accelerated luminal antigen presentation through a break in the epithelial layer, together with cytokines released from activated monocytes, might induce a second line of defense aiming at elimination of the massive antigen penetration into the mucosa . The postulated switch from secretory local immunity to a systemic type of local immune reaction could have different consequences:the local down-regulation of J chain in the IgA immunocytes could shift the production of polymeric IgA to monomers, jeopardizing secretory immunity; the disproportionate increase of IgG-producing cells could favor further inflammation and tissue damage through complement activation and arming of the killer cells, and cause autoimmune responses locally and in target organs at a distance (e.g . joint organs) . The discovery of subclinical gut inflammation in the SpA had therapeutic consequences . Sulphasalazine (SASP) has been proven to be an active drug in the treatment of IBD . Since the gut could play a crucial role in SpA, it was logic to use this drug in the treatment of this disease . Multiple open and double-blind studies have proven the effectiveness of this drug in SpA. Prog Nucleic Acid Res Mol Biol, 1996, 54, 101 - 25 Homologous genetic recombination in Xenopus: mechanism and implications for gene manipulation; Carroll D; Appropriately designed DNA substrates undergo very efficient homologous recombination after injection into the nuclei of Xenopus laevis oocytes . The requirements for this process are that the substrate be linear, that it have direct repeats to support recombination, and that these repeats be at or very near the molecular ends . Taking advantage of direct nuclear injection, the large amounts of DNA processed in a single oocyte, and the accessibility of recombination intermediates, we were able to analyze the mechanism of recombination in detail . Molecular ends are resected by a 5'-->3' exonuclease activity . When complementary sequences are exposed from two ends, they anneal . Continued 5'-->3' degradation removes the redundant strands; the 3' ends pair with their complements and can be extended by DNA polymerase to fill any gap left by the exonuclease . Joining of strands by DNA ligase completes the process . This mechanism is nonconservative, in that only one of the two original repeats is retained, and it has been dubbed single-strand annealing, or SSA . The capability for SSA accumulates during the later phases of oogenesis and persists into the egg . This pattern suggests that, like many activities of full-grown oocytes, SSA is stored for use during embryogenesis . The same or a very similar mechanism is prevalent in many other species, including bacteria, yeast, plants, and mammals, where it often provides the predominant mode of recombination of extrachromosomal DNA . Lessons learned about SSA are applicable to methods of gene manipulation . It is plausible that SSA has a normal function in the repair of double-strand breaks, but proof of this awaits identification of genes and enzymes uniquely involved in this style of recombination. EXS, 1996, 75, 433 - 49 Pharmacological aspects and therapeutic applications of lysozymes; Sava G; The therapeutic effectiveness of lysozyme (large scale manufactured hen egg-white lysozyme) is actually based on its ability to control the growth of susceptible bacteria and to modulate host immunity against infections and depressions of immune responses . If the former is based on the first evidence of the biological activity of this enzyme, the second is a relatively recent acquisition of extreme importance for the possibilities offered in terms of the regulation of the functioning of the host's immune system . Antibotic activity and immune stimulating effects are also used together, as in the case of the treatment of gastrointestinal infections, including those originated by therapeutical treatments . Based on these biological properties, in addition to the wide range of therapeutic activities for which lysozyme was exploited in the past, at present the most promising data concern the prevention of bacterial cariogenesis and treatment of cancer patients to improve the effectiveness of anticancer drugs or to allow the host to recover from the immune suppression caused by anticancer treatments . However, lysozyme does not yet hold a clear place as an immune modulating agent, in spite of the fact that it has been shown to stimulate immunity with no difference between experimental animals and human beings . The hope is therefore that doctors will understand its potential and that they will take advantage of the existence of this simple and useful molecule. EXS, 1996, 75, 365 - 409 alpha-Lactalbumins and lysozymes; McKenzie HA; Lysozyme is ubiquitous in a variety of tissues and secretions . Chick-type (c-type) lysozymes lyse the cell walls of certain bacteria . In contrast, alpha-lactalbumin appears to occur only in mammalian milk and colostrum . It has the unusual property of acting as a modifier protein to modify the action of galactosyl transferase to a lactose synthase . Both proteins have diverged from a common ancestor . This is reflected in the striking relationship between their amino acid sequences, and the high conservation of disulfide bridges, their intron-exon organization, and three-dimensional structures . In studying their evolutionary relationships some important differences are noted, e.g., all alpha-lactalbumins strongly bind Ca(II), but only some c-type lysozymes do so . These properties point the way to future investigations that are necessary before firm conclusions can be made about their evolutionary history. EXS, 1996, 75, 105 - 10 Lysozyme substrates; Holtje JV; The natural substrate of lysozyme is the rigid layer of bacterial cell walls, the murein (peptidoglycan), which is a gigantic polymer of (GlcNAc-MurNAc)n polysaccharide strands crosslinked through short peptide bridges at the lactyl groups of the muramic acid residues . Thus, lysozyme lyses bacteria by degrading their protective exoskeleton, the murein sacculus . The high molecular weight murein is thereby hydrolysed to low molecular weight muropeptides, a process that can be followed quantitatively by different methods . However, due to the insolubility of the murein sacculus, the enzyme kinetics are rather complex . Therefore, a variety of different low molecular weight substrates have been prepared, both murein degradation products and synthetic compounds . These substrates allow a better characterization of the binding and catalytic mechanism of lysozyme . In addition, they are used in various photometric, isotopic and immunological lysozyme assays. Rev Pneumol Clin, 1996, 52(3), 192 - 5 {Eikenella corrodens pleuro-pulmonary infection . Apropos of a case}; Salem M et al.; A case of single bacteria pleuropulmonary infection was observed in a 21-year-old patient with heterozygous sicle-cell amenia . The role of this bacteria in pleuropulmonary suppuration, the pathophysiology and secondary lesions are discussed together with therapeutic modalities. Zhonghua Yu Fang Yi Xue Za Zhi, 1996 Jan, 30(1), 23 - 5 {Comprehensive cost-benefit evaluation for the improvement of rural water supply in Hunan province}; Tang M et al.; Comprehensive cost-benefit evaluation for the improvement of rural water supply was conducted according to the national standard methods for analysis of water quality . Results showed total bacteria count and coliform group count in the water declined by 51% after the improvement, and synthetic index of water quality declined to 19% of that before the Improvement . Environmental epidemiological studies showed yearly incidence of hepatitis A, typhoid fever, diarrhea and enteritis decreased by 2.20/1,000, 0.39/1,000, 2.70/1,000 and 14.30/1,000, respectively, as compared with those before the improvement, and with the manpower capital method, economic loss caused by the above-mentioned diseases lowered by 37,238 yuan, economic benefit gained by saving labor-time for water-taking reached 198,644 yuan, and family income increased 164,188 yuan per year . It suggests the above indicators all can be used in comprehensive cost-benefit evaluation of the improvement of rural water supply. Probl Tuberk, 1996, (3), 12 - 4 {Tuberculosis in refugees from foreign countries}; Rybka LN et al.; Screening for tuberculosis has revealed active disease in 30 of 168 examinees . They have arrived in Russia as refugees from Somalia, Afghanistan, Angola and other countries . 24 refugees had pulmonary tuberculosis . Destruction of pulmonary tissues and massive discharge of M . tuberculosis were found in 10, 2 and 1 patients with infiltrative, disseminated and fibrous-cavernous tuberculosis . A specific therapy with 3-4 drugs stopped discharge of the bacteria with sputum . It is stated that refugees from foreign countries may constitute a great epidemiological threat of tuberculosis for contacting Russian population. Cytokine, 1996 Jan, 8(1), 14 - 20 Avian stem cell factor (SCF): production and characterization of the recombinant His-tagged SCF of chicken and its neutralizing antibody; Bartunek P et al.; Recombinant chicken stem cell factor (SCF) was produced in bacteria as a histidine-tagged protein (His delta SCF) and purified by affinity chromatography on an Ni(2+)-NTA agarose column . His delta SCF is devoid of signal peptide and transmembrane domains and therefore mimics a soluble form of SCF . We demonstrate that the factor is biologically fully active in supporting sustained proliferation of SCF-dependent red blood progenitor cells of chicken in vitro . Polyclonal SCF-specific antibodies were raised in rabbits and shown to display potent SCF-neutralizing activity . Thus, recombinant chicken His delta SCF together with the SCF-specific antibody provide valuable tools for studying the role of SCF and c-kit receptor in development, growth and differentiation of avian haematopoietic cells. Baillieres Clin Endocrinol Metab, 1996 Jan, 10(1), 9 - 27 The TSH receptor and thyroid diseases; Paschke R et al.; Recent advances in the understanding of the molecular biology of the TSH receptor have had a considerable impact on several aspects of thyroidology . The identification and functional characterization of mutations in the TSH receptor gene which constitutively activate the TSH receptor in the absence of its ligand provide an explanation for the molecular mechanism which is most likely responsible for the majority of the hyperfunctioning thyroid adenomas . Moreover, these constitutively activating mutations also cause a new form of familial hyperthyroidism: non-autoimmune autosomal dominant hyperthyroidism and also sporadic cases of congenital non-autoimmune hyperthyroidism . TSH receptor mutations which cause a reduced sensitivity to TSH have been identified as the cause of non-autoimmune congenital hypothyroidism . TSH receptor mRNA variants have been found in thyroid associated ophthalmopathy . If protein expression for these variants can be demonstrated, this finding could advance our understanding of thyroid associated ophthalmopathy . The ability to produce large quantities of TSH receptor protein in bacteria has led to the generation of more sophisticated assays for TSH receptor antibodies and enabled the generation of an animal model for thyroid autoimmunity. Arq Bras Cardiol, 1996 Jan, 66(1), 1 - 3 {Infective endocarditis in 100 patients subjected to heart transplantation}; Uip DE et al.; PURPOSE: An analysis of occurence, etiology, clinical aspects and death rate of infectious endocarditis cases involving patients who underwent heart transplantation . METHODS: 100 consecutive heart transplant patients were analysed; follow-up varied from three to 90 (medium of 25.38, SD +/- 27.97) months . Diagnostic criteria for endocarditis were those of the epidemiology and quality control service, Heart Institute, Hospital das Clinicas da Faculdade de Medicina da Universidade de Sao Paulo, that agree with those of the Center for Disease Control . Diagnosis was established mostly by blood cultures, echocardiograms, either transthoracic or esophageal and autopsy . RESULTS: Six cases of endocarditis were recognized, with four deaths; in three of those we found severe systemic compromise . All cases had fever as an important symptom . CONCLUSION: Death risk seems high of infectious endocarditis after heart transplantation . The cause of this high risk appears to be linked to the bacteria themselves, to association with other clinical situations and to the use of immunosuppresive agents . We recommend a high index of suspition when risk factors are present in order to make a rapid diagnosis soon enough in the natural history of the disease; treatment has to be started as soon as possible. J Comp Pathol, 1996 Jan, 114(1), 81 - 91 Relationship of acid phosphatase activity to ultrastructural features in mice inoculated with Mycobacterium paratuberculosis; Tanaka S et al.; Macrophage activation, measured as increased acid phosphatase (AcPase)-positive areas by image analysis, and ultrastructural features were examined in granulomatous mycobacterial lesions of mice innately susceptible (BALB/c mice; Bcg) and innately resistant (C3H/HeJ mice; Bcg) to Mycobacterium paratuberculosis strain ATCC 19698 . In the liver and spleen of BALB/c mice 3 weeks after intraperitoneal inoculation with M . paratuberculosis, AcPase activity detected in epithelioid cell nodules was high; it had decreased, however, in the liver and spleen after a further 3 and 6 weeks, respectively . In C3H/HeJ mice, the size of epithelioid cell nodules in the liver and spleen was smaller than in BALB/c mice, and infiltrating macrophages, which had increased by week 9 after inoculation, showed high AcPase activity . Ultrastructurally, by week 32 in BALB/c mice, small phagolysosomes (SPLs) had greatly increased in number in the epithelioid cells . These SPLs contained a few AcPase-positive areas and a small number of bacteria, most of which were surrounded by an electron-translucent space (or electron-transparent zone {ETZ}) . In contrast, only a few SPLs were observed in C3H/HeJ mice at week 32; in the liver and spleen, large phagolysosomes (LPLs) showed high AcPase activity and contained many degenerated bacteria, which also had an ETZ . These results suggest that the enzymatic and ultrastructural differences in phagolysosomes between BALB/c mice and C3H/HeJ mice reflect the susceptibility of these mouse strains to M . paratuberculosis. J Photochem Photobiol B, 1996 Jan, 32(1-2), 59 - 65 Ex-vivo treatment of gastric Helicobacter infection by photodynamic therapy; Millson CE et al.; Attempts to develop PDT for eradication of Helicobacter infection have only been successful in vitro . We have investigated the effect of topical sensitization (except ALA) of Helicobacter mustelae on explanted ferret gastric mucosa using one of five sensitizers (methylene blue (MB), toluidine blue O (TBO), phthalocyanine, haematoporphyrin derivative and 5-aminolavulinic acid), followed by irradiation with an appropriately tuned copper vapour pumped dye laser . A 90% reduction in counts of bacteria sensitized with 0.75 mg TBO kg-1 were seen after irradiation with 200 J cm-2 . Concentrations of MB of 0.75 mg kg-1 and 7.5 mg kg-1 were not toxic to H . mustelae, but the further addition of 20 J cm-2 laser light reduced colony counts by more than 99% . MB at a concentration of 75 mg kg-1 exhibited significant dark toxicity towards H . mustelae, but further addition of 20 J cm-2 laser light resulted in near eradication of all colonies . The remaining three compounds were ineffective . Finally, we studied the microscopic fluorescence distribution of MB (7.5 mg kg-1) on ferret gastric mucosa after topical administration . Fluorescence was greatest in the superficial mucosal layer, upon which lies the bacteria . However, from experiments on rats, the energy required to kill the sensitized bacteria was insufficient to damage the underlying mucosa . We conclude that Helicobacter can be killed on host mucosal epithelium following topical administration of MB and subsequent exposure to laser light. Scand J Gastroenterol Suppl, 1996, 214, 2 - 4; discussion 9-12 History of Helicobacter infection; Rune SJ; Helicobacter pylori infection is now recognized as the cause of most cases of chronic gastritis, and a major causal factor in peptic ulcer disease . However, though the presence of intragastric bacteria has been recognized for many years, the connection between H . pylori and peptic ulcers has only been established during the last decade . Pathological studies have suggested that intragastric bacteria damage the gastric mucosa, reducing its resistance to acid, while epidemiological studies have suggested that a non-physiological factor, such as bacterial infection, is involved in the development of peptic ulcer . Subsequently, H . pylori, associated with chronic gastritis, was identified in gastric biopsies from patients with peptic ulcer . Eradication of H . pylori has been shown to reduce the recurrence of peptic ulcer after ulcer healing, and such treatment is now widely accepted. Schweiz Arch Tierheilkd, 1996, 138(3), 115 - 20 {Identification and diagnosis of Taylorella equigenitalis by a DNA amplification method (PCR)}; Miserez R et al.; A polymerase chain reaction (PCR) for identification of Taylorella equigenitalis was developed . The oligonucleotide primers are based on the DNA sequence of the rrs gene of T . equigenitalis, encoding for the 16S ribosomal RNA . Analysis of 21 strains of T . equigenitalis from England, USA and Switzerland showed an amplification product of 410 bp with identical Sau3A restriction profile . The sensitivity of the PCR-Assay was estimated to detect 50 to 500 bacteria of T . equigenitalis in a mixture with frequently found contaminants . Further analysis of culture from 60 genital swabs, taken in the course of the control of the contagious equine metritis in horses and donkeys, of experimental assays as well as of two positive cases from the diagnostic showed that this PCR-assay can be used to identify and to detect strains of T . equigenitalis . In addition, preliminary results indicate that the method is also applicable for direct in vitro establishment of the presence of T . equigenitalis in clinical samples. Zhonghua Hu Li Za Zhi, 1996 Jan, 31(1), 5 - 7 {Study on the collecting method of human umbilical cord blood}; Cai WZ et al.; The methods of cord blood collection was investigated in this study . The purpose of this study was to evaluate human umbilical cord blood as a alternative to bone marrow in the provision of transplantable progenitor cells for hematopoietic reconstitution . The results showed that using modified technique with a sterile and closed system, the volume of cord and placental blood would all exceed 120 ml . The average amount of blood collected was 132.2 +/- 12.13 ml and the highest was 158 ml . Cord blood culture for bacteria and fungi were negative . Blood clots were not present . And in the examination of each single cord blood, the contents of nucleated cells (NS) and mono-nucleated (MNS) were, respectively, 18.9 X 10 +/- 1.7 X 10, 8.2 X 10 +/- 0.8 X 10 . The result suggested that the collection of placental blood with the modified technique in a sterile and closed system is a simple, safe and efficient procedure . The average amount of blood collected and hematopoietic cells would satisfy the needs of cord blood transplantation . Immediate cut of the umbilical cord followed by a prompt puncturing is critical for the volume and quality of the sample. Gene, 1996, 173(1 Spec No), 89 - 98 Green fluorescent protein marks skeletal muscle in murine cell lines and zebrafish; Moss JB et al.; The green fluorescent protein (GFP) acts as a vital dye upon the absorption of blue light . When the gfp gene is expressed in bacteria, flies or nematodes, green fluorescence can be directly observed in the living organism . We inserted the cDNA encoding this 238-amino-acid (aa) jellyfish protein into an expression vector containing the rat myosin light-chain enhancer (MLC-GFP) to evaluate its ability to serve as a muscle-specific marker . Transiently, as well as stably, transfected C2C12 cell lines produced high levels of GFP distributed homogeneously throughout the cytoplasm and was not toxic through several cell passages . Expression of MLC-GFP was strictly muscle-specific, since Cos 7 fibroblasts transfected with MLC-GFP did not fluoresce . When GFP and beta Gal markers were compared, the GFP signal was visible in the cytoplasm of the living cell, whereas visualization of beta Gal required fixation and resulted in deformation of the cells . When the MLC-GFP construct was injected into zebrafish embryos, muscle-specific gfp expression was apparent within 24 h of development . gfp expression was never observed in non-muscle tissues using the MLC-GFP construct . Transgenic fish continued to express high levels of gfp in skeletal muscle at 1.5 months, demonstrating that GFP is an effective marker of muscle cells in vivo. Gene, 1996, 173(1 Spec No), 5 - 11 Chemistries and colors of bioluminescent reactions: a review; Hastings JW; Many different organisms, ranging from bacteria and fungi to fireflies and fish, are endowed with the ability to emit light, but the bioluminescent systems are not evolutionarily conserved: genes coding for the luciferase proteins (Lase) are not homologous, and the luciferins are also different, falling into many unrelated chemical classes . Biochemically, all known Lase are oxygenases that utilize molecular oxygen to oxidize a substrate (a luciferin; literally the "light-bearing' molecule), with formation of a product molecule in an electronically excited state . The color of the light may differ, even though the same luciferin/Lase system underlies the reaction . Filters or differences in Lase structure are responsible in some cases; in others a secondary emitter associated with a second protein is involved . In the coelenterates a green fluorescent protein, whose chromophore is derived from the primary amino-acid sequence, results in a red shift of the emission . In the bacteria accessory proteins causing either blue- or red-shifts have been isolated from different species; the chromophores are noncovalently bound . Although radiationless energy transfer has been implicated in the excitation of such accessory emitters, this may not be so in all cases. Exp Gerontol, 1996 Jan-Apr, 31(1-2), 13 - 9 A prospective view on phosphatases and replicative senescence; Bernier L et al.; Addition of phosphate to proteins by kinases, or its removal by phosphatases, is probably the control mechanism most often used by cells to maintain homeostasis . This mechanism presents the advantage of being fast, versatile, and easily reversible . It is used by all organisms from bacteria to man . Although more is known about the kinases, recent studies are beginning to shed light on the role of phosphatases, the enzymes that are responsible for terminating the effects of phosphorylation . These enzymes are perfect candidates for controlling all the crucial check points during cell cycle traverse, and as such, will be found to be responsible for many important decision in the life of a cell, including entry into replicative senescence. Vet Microbiol, 1996 Jan, 48(1-2), 1 - 7 An experimental ovine foot abscess model using a Fusobacterium necrophorum biotype AB; Corner LA et al.; An experimental procedure is described for the production of foot abscess in sheep that mimics the natural disease . Lesions were produced by the intradermal inoculation of suspensions of Fusobacterium necrophorum biotype AB containing from 5 x 10(2) to 5 x 10(8) bacteria, into interdigital skin devitalized by freezing with liquid nitrogen . A dose of 5 x 10(5) bacteria induced the development of foot abscess in 3 of 4 and 8 of 8 inoculated feet . It was found that to produce foot abscess in devitalized tissue required between 10(3) and 10(6) fewer bacteria than were necessary to produce similar lesions in healthy tissue. Life Sci, 1996, 59(3), 219 - 25 Constitutive expression and structural diversity of inducible isoform of nitric oxide synthase in human tissues; Park CS et al.; Nitric oxide (NO) produced by inducible nitric oxide synthase (iNOS) plays a major role in immune responses to bacteria and tumors, and the gene is induced by endotoxin and cytokines . However, we have detected iNOS cDNA sequences expressed constitutively at low level from human retinal, cerebellar and skeletal muscle tissues using northern-blot and RT-PCR analyses . In northern-blot analysis, two types (4.5 kb and 4.2 kb) of iNOS mRNA have been observed in retinal tissue, whereas only one type of mRNA was observed in cerebellum (4.5 kb) and skeletal muscle (4.2 kb) . This result indicates that the presence of differential expression and/or structural diversity of the iNOS gene in various tissues, and some cells can express iNOS gene constitutively . We have also demonstrated a structural diversity formed by alternative splicing in the open reading frame sequence of the iNOS cDNA cloned from retinal tissue, which may reflect functional differences of iNOS gene. Microb Pathog, 1996 Jan, 20(1), 31 - 40 Accelerated development of genital Chlamydia trachomatis serovar E in McCoy cells grown on microcarrier beads; Wyrick PB et al.; Chlamydia trachomatis serovar E is a major cause of bacterially-acquired sexually transmitted infections . Stock cultures of these obligate intracellular bacteria are often propogated in McCoy cells . We recently reported that greater infectious titers of chlamydiae could be obtained if the McCoy cells were cultured on collagen-coated microcarrier beads versus plastic flasks, although the reason for the difference in efficiency was not clear . This study analyzed the development of C . trachomatis grown in McCoy cells by the two methods . Transmission electron microscopy analysis revealed an accelerated chlamydial development, with maturation of reticulate bodies into elementary bodies sooner in McCoy cells grown on the porous substratum . Comparison of particle counts versus infectivity titers indicated the production of fewer numbers of elementary bodies but which were highly infectious sooner from the infected McCoy cell-microcarrier bead cultures than from duplicate infected McCoy cell cultures grown in plastic tissue culture flasks. Rev Stomatol Chir Maxillofac, 1996, 97(2), 121 - 4 {Transmission of infection in oral medicine . Evaluation of the risk of transmission in the office surgery}; Vacher C et al.; The present study was designed to estimate the risk of transmission of infectious diseases by dental equipment in stomatology . The protocol was carried out on 30 patients . The three following samples were collected from each patient: (a) oral flora before any treatment, (b) aerosol produced by the hand-piece before internal decontamination, (c) aerosol produced by the hand-piece after internal decontamination . The results of this study indicate there was a risk of transmission of infectious diseases in 3 cases (10%) with dental equipment, even if there were no consequences for healthy individuals . We have to improve the security in stomatology, with internal and external decontamination after each patient. Pediatr Med Chir, 1996 Jan-Feb, 18(1), 109 - 12 {Pertussis . Critical revision of its physiopathology . Implications}; Fiore D; The re-examination of Bordetella pathogenic power and of the mechanisms which regulate relations between these bacteria and men make us conclude that in pertussis and with intramuscular vaccines an antihistaminic "coverage" is needed and antipertussis vaccinations is to be made by natural way . Via aerosol, minor intrinsic toxicity of acellular vaccines will be determinant. Khirurgiia (Mosk), 1996, (1), 52 - 6 {New antiseptics and disinfectants in surgery (2)}; Pkhakadze TYa et al.; Various forms of plivasept, an agent produced by Pliva (Croatia), were studied in the laboratory and clinic and found to be highly effective . For the prevention of intrahospital infections the use of 0.02% and 0.05% water solutions of plivasept without PAV is advisable as an antiseptec for inyracavitary administration, 0.05% water solution of plivasept for disinfection of surfaces and equipment, 0.5% alcohol plivasept solution for disinfection of instruments, a plivasept tincture for surgical treatment of the skin on the hands and the injection and operative fields, and foam-forming plivasept for hygienic treatment of the hands. Antonie Van Leeuwenhoek, 1996 Jan, 69(1), 15 - 23 Simulation of constituent processes of anaerobic degradation of organic matter by the "methane" model; Vavilin VA et al.; The model of anaerobic digestion described earlier by the authors was used for analysis of the different phases of the process . It was shown that at the glucose conversion a coexistence of hydrogen-producing acidogenic bacteria and hydrogen-utilizing non-methanogenic bacteria causes a hydrogen partial pressure decrease at an increase of solids retention time (i), the intensity of the negative feed-back effect in sulfate-reduction through hydrogen sulfide formation is regulated by the pH level during an oscillation dynamics in acetate/sulfate system (ii), under the toxicity influence the processes of methanogenesis and acetogenesis together with hydrolysis may be rate-limiting steps in the anaerobic system with particulate substrate degradation (iii). Schweiz Med Wochenschr Suppl, 1996, 76, 21S - 27S The use of aminoglycosides in neutropenic patients; Cometta A et al.; Aminoglycoside antibiotics are widely used for the empirical treatment of febrile neutropenic patients . They are administered in combination with beta-lactam antibiotics and sometimes with glycopeptide antibiotics . This review sets out to determine which patient populations are most likely to benefit from the administration of aminoglycoside antibiotics . In addition, the efficacy and safety of aminoglycoside antibiotics administered in a single daily dose are discussed in the light of a recent trial. J Dairy Sci, 1996 Jan, 79(1), 98 - 104 Effects of forage percentage and canola seed on ruminal protein metabolism and duodenal flows of amino acids in steers; Hussein HS et al.; The objective was to determine the effects of dietary forage percentage and fat supplementation on ruminal N metabolism, duodenal flows of AA, and digestion of N . Six ruminally and duodenally cannulated steers were offered six isonitrogenous diets for ad libitum intake twice daily in a 6 x 6 Latin square design . Treatments were arranged as a 2 x 3 factorial with two forage percentages (70 vs . 30% of dietary DM as corn silage) and three forms of canola seed supplementation, including no canola seed or canola seed added at 10% of dietary DM as whole seed treated with alkaline H2O2 or as crushed seed . No interactions between dietary forage percentage and canola seed supplementation occurred for any of the measurements . Duodenal flows of NAN and AA were greater for diets containing low forage than for diets containing high forage . However, duodenal flows of total N and NAN did not differ among diets when corrected for differences in intake . Efficiency of bacterial protein synthesis and duodenal flows of bacterial N and AA were increased when treated whole canola seed was supplemented . Apparent total tract digestibility of N was not altered by dietary forage percentage or canola seed supplementation . Results indicate that fat supplementation from canola seed (at 5% of dietary DM), in either form, had no effects on ruminal N metabolism or flows of AA to the duodenum and suggest that treated whole canola seed may stimulate ruminal bacterial protein synthesis. Rocz Akad Med Bialymst, 1996, 41(1), 129 - 35 Unknown species of rickettsiae isolated from Ixodes ricinus tick in WaÅ‚cz; Kruszewska D et al.; Ticks Ixodes ricinus have been harvested in park down-town Walcz in June 1994 . Presence of Borrelia burgdorferi and rickettsiae has been tested with PCR . B . burgdorferi and Coxiella burnetii have not been detected in any tick whereas characteristic for other rickettsiae DNA fragment of gene encoding citrate-synthase has been found . Bacterial strain has been cultured from the tick . Biochemical properties of isolated strain has strongly suggested that these bacteria belong to genus Bartonella (Rochalimaea) . Further identification of bacterial DNA with RFLP-PCR (restriction fragment length polymorphism-PCR) has shown characteristics of Bartonella bacilliformis species not recognised in Poland until recently. Immunology, 1996 Jan, 87(1), 71 - 9 Bovine gamma/delta T-cell proliferation is associated with self-derived molecules constitutively expressed in vivo on mononuclear phagocytes; Okragly AJ et al.; Bovine gamma/delta T cells have been shown previously to proliferate when cocultured with gamma-irradiated bovine monocytes in the 'autologous mixed leucocyte reaction' (AMLR) . It was suggested that the response may be to culture-derived or culture-induced antigenic epitopes . Data presented here indicate that the gamma/delta T-cell stimulatory activity is attributable to a self-derived cell-surface molecule of mononuclear phagocytes that is constitutively expressed in vivo . The ability to induce an AMLR did not require in vitro culture or stress associated with in vitro isolation of cells or increased temperature since it could be induced by monocytes fixed by paraformaldehyde during blood collection from normal animals . Furthermore, stimulation by monocytes did not depend upon secreted molecules since fixed monocytes that had been incubated overnight at 37 degrees to allow secretion of preformed molecules, or subjected to hypotonic shock in H2O for 10 min before addition to the cultures, induced an AMLR as did plasma membranes prepared from ex vivo monocytes . In contrast, enzymatic treatment of monocytes to digest surface molecules followed by fixation destroyed their ability to stimulate an AMLR . The ability of monocytes to stimulate proliferation of gamma/delta T cells was distinguishable from their ability to stimulate alpha/beta T cells, since the former was destroyed by glutaraldehyde fixation whereas stimulation of alpha/beta T cells by major histocompatibility complex (MHC)-presented antigenic epitopes is not . Moreover, induction of proliferation of bovine gamma/delta T cells was not MHC-restricted . Finally, bovine alveolar macrophages, sheep monocytes and transformed bovine monocytes stimulated proliferation of bovine gamma/delta T cells whereas none of the following did so: human monocytes, murine macrophages, bovine myeloid cells other than mononuclear phagocytes, other nucleated cells found in bovine blood including activated MHC class II-bearing B cells, and a variety of species of bacteria . Thus, the stimulatory epitope is unique to and conserved among mononuclear phagocytes of ruminants . Demonstration of stimulation of bovine gamma/delta T cells by self-derived molecules is consistent with reports for murine gamma/delta T cells. Probl Tuberk, 1996, (2), 6 - 7 {Role of occupational factors in the incidence and course of respiratory system tuberculosis in fishery workers in the South Far East}; Medvedev VI et al.; Tuberculosis morbidity among fishery workers in the South of Russian Far East remains high . Moreover, it is higher in fishermen working in the sea than in those who are engaged in fish-factories . This conclusion on fishing as a factor of tuberculosis risk was made basing on the results of 91 patients with tuberculosis of the respiratory organs . 65.9% of them were members of fishing ship's crew (group 1), 34.1% were workers of fish-factories (group 2) . Pulmonary infiltrative tuberculosis running with exudation was typical for both groups, but group 1 subjects developed destruction of pulmonary tissue and discharged bacteria significantly more frequently . They had more compromised cellular immunity. Probl Tuberk, 1996, (2), 13 - 6 {Characteristics of the course and treatment of pulmonary tuberculosis associated with nonspecific respiratory diseases in children}; Mitinskaia LA et al.; Specific features of intrathoracic tuberculosis course, outcomes and treatment are outlined for 109 children . In 59 of them tuberculosis was associated with nonspecific respiratory diseases (NRD) . 28.8%, 25.4%, 18.6, 27.2% of patients had cystic hypoplasia, chronic hypoplasia, recurrent pneumonia, recurrent bronchitis, respectively . Complicated course of tuberculosis occurred 2 times more frequently in its combination with NRD (71.2%) . Destruction and discharge of bacteria were recorded in 49.2 and 47.5% of patients, respectively . Undulating running was 3.4 and side effects 1.6 times more frequent . Tuberculous children with NRD need longer antituberculous therapy using wide-spectrum antibiotics, symptomatic and exercise treatment, massage, surgical intervention if necessary . Complete resolution of lung lesions in NRD children were seen 4.8 times less frequently . In case of late diagnosis 54% of them retained residual changes in the form of lung tissues fibrosis, calcified foci in the lungs and lymph nodes. J Dent Res, 1996 Jan, 75(1), 529 - 34 Michael Buonocore and the Eastman Dental Center: a historic perspective on sealants; Handelman SL et al.; Dr . Michael Buonocore is known for his innovative research on the preparation of the enamel surface with a weak acid to enhance adhesion of an organic plastic chemical sealant and the polymerization in situ of a sealant with ultraviolet light . His co-workers at Eastman Dental Center aided and extended his research findings . The purpose of his original research was the development of a sealant to prevent occlusal caries on posterior teeth . However, the major impact of his work has been the development of adhesive restorative techniques . Although it has been demonstrated that (i) bacteria tend to die out and caries does not progress if early caries lesions are inadvertently sealed, (ii) sealant retention rates are favorable, and (iii) sealants are cost-effective, the use of sealants by the profession is still far short of early expectations. Gerontology, 1996, 42(1), 40 - 5 Are hydrogen breath tests valid in the elderly? Mac Mahon M, Gibbons N, Mullins E, O'Moore RR, Keane CT, Walsh JB, Coakley D. Hydrogen breath testing (HBT) is frequently used as an alternative to small bowel aspiration in the diagnosis of small intestinal bacterial overgrowth (SIBO) . The role of the glucose HBT was assessed in 30 elderly patients . A positive HBT was recorded in 15 of 20 SIBO cases and 7 of 10 culture negatives (sensitivity 75% and specificity 30%) . The correlation coefficients between hydrogen gas (H2) rise and total bacterial count (r = 0.21) and H2 rise and anaerobic count (r = 0) were not significant . Fasting H2 levels were raised in only 4 of the 20 SIBO cases . This study indicates that the HBT is not reliable in the diagnosis of SIBO in the elderly . There was no evidence from the data that different H2 levels or bacterial counts would significantly alter the reliability of the HBT . This work suggests that factors other than small bowel bacteria are involved in the production and expiration of H2 in the elderly, and that these factors need to be considered in the interpretation of this breath test. Eur J Clin Microbiol Infect Dis, 1996 Jan, 15(1), 71 - 5 Impact of using an indwelling introducer on diagnosis of Swan-Ganz pulmonary artery catheter colonization; Valles J et al.; A prospective study was conducted to determine how the use of an indwelling introducer influences the diagnosis of pulmonary artery catheter (PAC) colonization . Sixty-six consecutive PACs and introducers were aseptically removed over a 15-month period . Two segments of the catheter and the catheter tip (a proximal segment from the portion of the catheter beneath the introducer) and two segments of the introducer (a proximal intradermal segment and the introducer tip) were cultured using a semiquantitative technique . Nineteen of 66 (28.7%) PACs showed colonization, representing an incidence of 5.6 episodes per 100 catheterization-days . Catheter tip cultures identified only 68% of colonized PACs; this yield rose to 91% when introducer tip cultures were added . These results indicate a need to evaluate both introducer tip cultures and catheter tip cultures for an accurate diagnosis of PAC colonization when an indwelling introducer is used. Cytogenet Cell Genet, 1996, 72(4), 299 - 302 A third P-domain peptide gene (TFF3), human intestinal trefoil factor, maps to 21q22.3; Schmitt H et al.; Small peptides displaying a cysteine-rich module (termed P-domain or trefoil motif) form a recently increasing group of peptides abundantly expressed at mucosal surfaces of specific tissues and are associated with the maintenance of surface integrity . The estrogen-inducible pS2 gene (BCEI) and the human homolog to the porcine spasmolytic peptide (hsP) gene (SML1) appear synchronously expressed in healthy stomach mucosa and several carcinomas of the gastrointestinal tract . Both genes were shown to be located at 21q22.3 . A new trefoil peptide from human intestinal mucosa (hITF/hP1.B) and its gene (TFF3) were described recently . By PCR analysis of a somatic cell hybrid panel and FISH using two large genomic recombinants (110 kb, 210 kb) cloned in the Bacterial Artificial Chromosome (BAC) system, we show that this gene coding for the new member of human P-domain/trefoil peptides also maps to chromosome region 21q22.3 suggesting a physical linkage of all three trefoil peptide genes. Arch Virol, 1996, 141(2), 209 - 18 Frequency of morphological phage descriptions in 1995; Ackermann HW; At least 4500 bacterial viruses have been examined in the electron microscope since 1959 . About 4400 phages (96%) are tailed and only 162 phages (4%) are cubic, filamentous, or pleomorphic . Phages belong to 12 virus families and occur in about 130 bacterial genera . Phages are listed by morphotypes and host genera . Siphoviridae or phages with long, noncontractile tails include about 60% of tailed phages. Curr Opin Periodontol, 1996, 3, 3 - 9 Epidemiology of periodontal diseases; Beck JD et al.; This review summarizes findings from 46 reports selected from over 130 epidemiologic studies published in 1993 and 1994 . Recent findings from longitudinal studies are consistent with the concept that periodontal destruction proceeds in random bursts at specific sites and demonstrate that some individuals have an elevated risk of attachment loss . These and other studies provide further evidence that cigarette smoking is a significant risk factor . New studies have quantified the risk of tooth loss due to periodontal disease, and there is better knowledge about HIV-associated periodontal diseases . Familial studies suggest that both environmental and genetic factors contribute to individual variations in etiologic factors (such as plaque) and periodontal diseases . Several studies propose an intriguing link between periodontal diseases and coronary heart disease, which may be mediated through risk factors common to both diseases, and as a direct consequence of the contribution of periodontal bacteria and their products to atherosclerosis. J Antibiot (Tokyo), 1996 Jan, 49(1), 50 - 3 Herquline B, a new platelet aggregation inhibitor produced by Penicillium herquei Fg-372; Enomoto Y et al.; A new herquline analog, herquline B was isolated from the culture broth of Penicillium herquei Fg372 . Herquline B contains one piperazine and two cyclohexenones . The pyrrolidine ring of herquline A was cleaved to yield herquline B . The IC50 value of herquline B against platelet aggregation induced by ADP and platelet-activating factor were 1.6 and 5.0 microM, respectively. Life Sci, 1996, 58(2), PL 41 - 6 Agmatine treatment is neuroprotective in rodent brain injury models; Gilad GM et al.; Agmatine is a naturally occurring guanidino compound, found in bacteria and plants, with several proposed nervous system-related functions suggestive of beneficial effects in central nervous system injury . Here evidence is presented that agmatine can exert potent neuroprotection in both in vitro and in vivo rodent models of neurotoxic and ischemic brain injuries . The cumulative evidence lead us to suggest that agmatine, a relatively nontoxic compound, be tried for potential therapeutic use after neurotrauma and in neurodegenerative disorders. J Cancer Res Clin Oncol, 1996, 122(4), 199 - 206 DNA-repair methyltransferase as a molecular device for preventing mutation and cancer; Sekiguchi M et al.; Alkylation of DNA at the 0(6) position of guanine is regarded as one o f the most critical events leading to induction of mutations and cancers in organisms . Once 0(6)-methylguanine is formed, it can pair with thymine during DNA replication, the result being a conversion of the guanine.cytosine to an adenine.thymine pair in DNA, and such mutations are often found in tumors induced by alkylating agents . To counteract such effects, organisms possess a mechanism to repair 0(6)-methylguanine in DNA . An enzyme, 0(6)-methylguanine-DNA methyltransferase, is present in various organism, from bacteria to human cells, and appears to be responsible for preventing the occurrence of such mutations . The enzyme transfers methyl groups from 0(6)-methylguanine and other methylated moieties of the DNA to its own molecule, thereby repairing DNA lesions in a single-step reaction . To elucidate the role of methyltransferase in preventing cancers, animal models with altered levels of enzyme activity were generated . Transgenic mice carrying the foreign methyltransferase gene with functional promoters had higher levels of methyltransferase activity and showed a decreased susceptibility to N-nitroso compounds in regard to liver carcinogenesis . Mouse lines deficient in the methyltransferase gene, which were established by gene targeting, exhibited an extraordinarily high sensitivity to an alkylating carcinogen. Eur J Haematol, 1996 Jan-Feb, 56(1-2), 54 - 61 Radioprotective effects of combination broncho-vaxom, a macrophage activator, and indomethacin, an inhibitor of prostaglandin production: relationship to myelopoiesis; Fedorocko P et al.; The effects of the bacterial extract broncho-vaxom (BV; radioprotective immunomodulator; 500 micrograms/mouse i.p., -24 h) and indomethacin (INDO; inhibitor of prostaglandin production; 2 x 40 micrograms/mouse i.m., -24 h and -3 h) on the post-irradiation recovery of hemopoietic functions in mice were investigated . Both agents were administered either alone or in combination . Endogenous spleen colony formation was increased in all treatment groups, with combination-treated mice exhibiting the greatest effects . Similarly, 24 h after combined administration of BV and INDO (i.e . at the time of presumed irradiation) to the non-irradiated mice granulocyte-macrophage colony-forming cell (GM-CFC) numbers were greater in the bone marrow and spleen . Also, as determined by hydroxyurea injection, there was an increase in the number of GM-CFC in the S-phase of the cell cycle in the bone marrow . However, GM-CFC in the spleen of combination pretreated mice was not stimulated to significant proliferation as compared to GM-CFC in the spleen of mice injected with BV alone . Combined modality treatment was also more effective than single agent treatments in accelerating bone marrow cellularity and GM-CFC regeneration, but not in accelerating GM-CFC regeneration in the spleen . Combined administration of BV and INDO to mice prior to lethal irradiation exerted an additional radioprotective effect and protected 95% of the C57Bl/6 mice. Nucleic Acids Res, 1996 Jan 1, 24(1), 92 - 7 Database on the structure of large ribosomal subunit RNA; De Rijk P et al.; Our database on large ribosomal subunit RNA contained 334 sequences in July, 1995 . All sequences in the database are aligned, taking into account secondary structure . The aligned sequences are provided, together with incorporated secondary structure information, in several computer-readable formats . These data can easily be obtained through the World Wide Web . The files in the database are also available via anonymous ftp. Nucleic Acids Res, 1996 Jan 1, 24(1), 26 - 8 The metabolic pathway collection from EMP: the enzymes and metabolic pathways database; Selkov E et al.; The Enzymes and Metabolic Pathways database (EMP) is an encoding of the contents of over 10 000 original publications on the topics of enzymology and metabolism . This large body of information has been transformed into a queryable database . An extraction of over 1800 pictorial representations of metabolic pathways from this collection is freely available on the World Wide Web . We believe that this collection will play an important role in the interpretation of genetic sequence data, as well as offering a meaningful framework for the integration of many other forms of biological data. Nucleic Acids Res, 1996 Jan 1, 24(1), 236 - 7 The Ribonuclease P Database; Brown JW; Ribonuclease P is the endoribonuclease responsible for the removal of leader sequences from tRNA precursors . Ribonuclease P is a ribonucleoprotein, and in bacteria the RNA alone is capable of pre-tRNA processing in vitro, i.e . it is a catalytic RNA . The Ribonuclease P Database is a compilation of ribonuclease P sequences, sequence alignments, secondary structures, three-dimensional models and accessory information, in the form of a hypertext document available via the Worldwide Web. Lett Appl Microbiol, 1996 Jan, 22(1), 21 - 5 Modified procedure for recovery of Giardia cysts from diverse water sources; Bielec L et al.; A compact battery-operated, portable field filtration apparatus was developed for efficient recovery of parasites, including Giardia lamblia cysts, from large volumes of water . In conjunction with modified filter elution and processing procedures, the filtration device was evaluated in field trials . To monitor performance of the procedure, a known number of Giardia lambia cysts were added during filtrations through a control injection assembly . This allowed calculation of cyst recovery at each test site, thereby allowing comparison of percentage recoveries from many diverse water sources . This apparatus and modified method now provide the tools to investigate further extraction and purification techniques to improve overall yield of Giardia lamblia, other parasites and potentially other organisms such as bacteria and viruses. Phytochemistry, 1996 Jan, 41(1), 7 - 22 Bioactive acylphloroglucinol derivatives from Eucalyptus species; Ghisalberti EL; The acyphloroglucinol derivatives produced by Eucalyptus species are reviewed . Aspects of their chemistry, stereochemistry, biological activity and biogenesis are discussed. Nippon Rinsho, 1996 Jan, 54(1), 67 - 72 {Cariostatic mechanisms of fluoride and its effects on human beings}; Sato T et al.; Fluorine is ubiquitous in man's environment, rocks, soil, water, air, etc, and mainly is in the form of inorganic fluoride (compounds of fluorine) and utilized extensively for industrial and medical purposes . It is also known that fluoride has both beneficial and toxic effects in man . One of the most beneficial effects of fluorine on human health is the application of fluoride solution for the prevention or reduction the incidence of dental caries . The major cariostatic mechanisms is believed to be that fluoride reduces dental caries through influencing the morphology of teeth by reducing the solubility of the enamel and promoting remineralization, and through its effect on oral cariogenic bacteria . Recently, the possible beneficial effects of fluoride on osteoporosis have been studied . Chronic effects of fluoride, described by many epidemiological studies, are only dental fluorosis and skeletal fluorosis . Most cases of acute poisoning in human subjects have been associated with the suicidal, accidental or experimental (by researcher himself) ingestion of a fluoride solution. Int J Syst Bacteriol, 1996 Jan, 46(1), 329 - 31 Species-specific sequences at the omp2 locus of Brucella type strains; Ficht TA et al.; A DNA sequence analysis of the omp2 locus of Brucella type strains revealed nucleotide differences that can be used for species identification . We developed specific probes which were used to verify the observed differences among the type strains following PCR amplification of portions of the omp2 locus. Appl Environ Microbiol, 1996 Jan, 62(1), 61 - 6 Aliphatic and chlorinated alkenes and epoxides as inducers of alkene monooxygenase and epoxidase activities in Xanthobacter strain Py2; Ensign SA; The inducible nature of the alkene oxidation system of Xanthobacter strain Py2 has been investigated . Cultures grown with glucose as the carbon source did not contain detectable levels of alkene monooxygenase or epoxidase, two key enzymes of alkene and epoxide metabolism . Upon addition of propylene to glucose-grown cultures, alkene monooxygenase and epoxidase activities increased and after an 11-h induction period reached levels of specific activity comparable to those in propylene-grown cells . Addition of chloramphenicol or rifampin prevented the increase in the enzyme activities . Comparison of the banding patterns of proteins present in cell extracts revealed that polypeptides with molecular masses of 43, 53, and 57 kDa accumulate in propylene-grown but not glucose-grown cells . Pulse-labeling of glucose-grown cells with {35S}methionine and {35S}cysteine revealed that the 43-, 53-, and 57-kDa proteins, as well as two additional polypeptides with molecular masses of 12 and 21 kDa, were newly synthesized upon exposure of cells to propylene or propylene oxide . The addition to glucose-grown cells of a variety of other aliphatic and chlorinated alkenes and epoxides, including ethylene, vinyl chloride (1-chloroethylene), cis- and trans-1,2-dichloroethylene, 1-chloropropylene, 1,3-dichloropropylene, 1-butylene, trans-2-butylene, isobutylene, ethylene oxide, epichlorohydrin (3-chloro-1,2-epoxypropane), 1,2-epoxybutane, cis- and trans-2,3-epoxybutane, and isobutylene oxide stimulated the synthesis of the five propylene-inducible polypeptides as well as increases in alkene monooxygenase and epoxidase activities . In contrast, acetylene, and a range of aliphatic and chlorinated alkanes, did not stimulate the synthesis of the propylene-inducible polypeptides or the increase in alkene monooxygenase and epoxidase activities. Mol Pharmacol, 1996 Jan, 49(1), 49 - 57 Palytoxin-induced Na+ influx into yeast cells expressing the mammalian sodium pump is due to the formation of a channel within the enzyme; Redondo J et al.; Palytoxin forms ionic channels in animal cell membranes but does not have similar effects on bacteria or yeast cells . These channels appear to be associated with the sodium pump . Using a heterologous expression system for the mammalian sodium pump in the yeast Saccharomyces cerevisiae, we recently demonstrated palytoxin-induced K+ efflux from yeast cells . Using the same system, we now show that the palytoxin-induced Na+ influx measured by others in animal cells is also directly associated with the sodium pump . Under the influence of palytoxin, yeast cells that express the mammalian sodium pump exchange extracellular Na+ ions for intracellular K+ ions with a stoichiometry of approximately 1:1 . Both fluxes can be inhibited by ouabain . K+ efflux can also be observed when extracellular Na+ is replaced by Li+, Cs+, or NH4+ . These data suggest that all palytoxin-induced ion fluxes measured so far in various cell systems are directly associated with the sodium pump . Palytoxin-induced Na+ influx or K+ efflux does not occur with yeast cells that express a truncated form of the sodium pump that is missing 44 of the carboxyl-terminal amino acids of the alpha 1 subunit . Scatchard analysis reveals only a slightly lower affinity of the truncated form for {3H}ouabain compared with the affinity of the native enzyme . Yeast cells expressing the truncated enzyme can bind {3H}ouabain, which can be displaced by palytoxin . Therefore, the inability of the truncated form to conduct ions under the influence of palytoxin is not due to the removal of the palytoxin binding site but rather to the removal of a part of the enzyme that participates in a direct or indirect way in the formation of the palytoxin-induced channel . Based on these findings, we conclude that palytoxin opens a channel within and not merely in the vicinity of the sodium pump . This might be the same channel that under normal conditions actively transports Na+ and K+ ions. Curr Top Microbiol Immunol, 1996, 210, 167 - 79 Structure and function of natural antibodies; Casali P et al.; Natural antibodies arise independently of known antigenic stimulation, are mostly IGM, polyreactive, and are generally encoded by V genes in germline configuration . Polyreactive IgM natural antibodies are produced by mainly B-1 cells which account for most of the B cell repertoire in the fetus and neonate, and possibly play a major role in the development and physiology of the human B cell repertoire . Although endowed with self-reactivity, natural antibodies also bind exogenous antigens {73, 74} . Exposure to environmental antigens is not necessary for the emergence of natural antibody-producing cell precursor clones to exogenous antigens, as suggested by the significant population of B cells capable of producing antibodies to a variety of bacterial antigens in germ-free animals . Because of their ability to bind a variety of exogenous antigens, including those on bacteria and viruses, natural antibodies play a major role in the primary line of defense against infections . A central issue related to the understanding of the physiopathologic roles of natural antibodies is whether precursors of cells producing natural antibodies, B-1a and B-1b lymphocytes, are capable of undergoing an antigen-driven clonal selection process, thereby producing autoantibodies with a high affinity for the selecting antigen . In this respect, we have clearly established that B-1 cells can express a hypermutation mechanism similar to that of conventional (B-2) cells . Furthermore, we have shown by gene shuffling, site-directed mutagenesis, and in vitro human Ig gene expression, that the main structural correlate for antibody polyreactivity is provided by the somatically generated H chain CDR3 . We have also shown that this Ig V region provides the main structural correlate for antigen-binding in monoreactive antigen-induced autoantibodies . These findings in the human are at the basis of our proposed structure-function model in which the antigen binding features of the germline template antibody are dictated by the somatically generated H chain CDR3, and perhaps, but at a lower degree, L chain CDR3; the point-mutation changes underlying the antigen-driven affinity maturation process would impact mainly the Ig V gene encoded segments . This structure-function model is being tested in our laboratory by analyzing the antigen binding activity of somatically mutated polyreactive autoantibodies that have been structurally reverted to their original putative unmutated configuration . Precise identification of the Ig gene and/or somatic recombination products mediating recruitment of unmutated B cell clonotypes, as well as those that are preferentially modified by an antigen-dependent selection process, should further our understanding of the mechanisms that shape the B cell repertoire in development and disease. Br J Haematol, 1996 Jan, 92(1), 200 - 11 The thromboxane mimetic, U-46619, induces the aggregation of fish thrombocytes; Hill DJ et al.; Fish blood contains nucleated cells termed thrombocytes which are thought to be functionally analogous to mammalian platelets . The present study was undertaken to assess the aggregatory response of peripheral blood thrombocytes from the rainbow trout, Oncorhynchus mykiss . Incubation of Percoll density gradient purified thrombocytes with the thromboxane A2 mimetic, U-46619 (0.03-10 microM), alone or in the presence of human or trout fibrinogen, elicited a dose-dependent aggregatory response . A greater amount of aggregation was observed in the presence of fish, rather than human, fibrinogen (e.g . c 55% compared with 15% maximal aggregation respectively) . This response was inhibited by pre-incubation of thrombocytes with the specific thromboxane A2 receptor antagonist, GR32191 (0.01-10 microM) with an IC50 value of 5.7 microM . Ultrastructural and aggregometry studies of thrombocyte aggregation at various time periods (1-12 min) after incubation with U-46619 (0.5 microM) revealed clear differences between the amount and dynamics of thrombocyte clumping in the presence of human, compared with trout, fibrinogen (final concentration 400 micrograms ml-1) . Thrombocytes rapidly underwent shape change and aggregation after only 1 min incubation without the initial involvement of any other cell types . The maximum degree of aggregation was achieved after 4-8 min with larger aggregates formed in the presence of trout compared than human fibrinogen . Ultrastructurally, the thrombocytes in these aggregates displayed a number of changes compared with non-stimulated cells, including increased pseudopodial activity, a more pronounced canalicular system and condensation of the nuclear heterochromatin . After 12 min incubation the clumps of thrombocytes showed progressive disaggregatory behaviour, with some cells reverting back to their normal in vivo appearance . Neutrophilic granulocytes, present as contaminants in the thrombocyte suspensions, were occasionally found attached to the thrombocyte aggregates, perhaps suggesting a specific interaction similar to that occurring in mammals . Finally, and also of some significance, was the finding that trout thrombocytes exhibit phagocytic activity in vitro towards cell debris and contaminating bacteria, indicating that this cell type may possess both haemostatic and immunological capacities. Urology, 1996 Jan, 47(1), 129 - 34 Bilateral emphysematous pyelonephritis: a case report and review of the literature; Stein JP et al.; Emphysematous pyelonephritis is a rare, rapidly progressive, life-threatening infection of the renal parenchyma . It most commonly is unilateral, is found almost exclusively in diabetics, is associated with gas-forming coliform bacteria, and is characterized by the presence of gas within the renal parenchyma . Early aggressive therapy (combined medical and surgical) is the hallmark of successful treatment . A high index of suspicion coupled with radiographic imaging is essential to make a timely diagnosis and guide therapeutic intervention . Bilateral emphysematous pyelonephritis is an even more rare phenomena, with only 14 reported cases in the English literature and is associated with a high patient mortality . We herein present an additional case and review the literature as it pertains to bilateral emphysematous pyelonephritis . Appropriate care requires aggressive combined medical and surgical therapy in an attempt to preserve renal function without increased mortality. Ugeskr Laeger, 1996 Jan 1, 158(1), 34 - 7 {Diagnosis and treatment of conjuncitivitis}; Ehlers N et al.; Conjunctivitis is a frequent reason for consulting a doctor . Most often treatment consist of antibiotics, even though sensitive bacteria are only rarely demonstrated . In our part of the world conjunctivitis is self-limiting, and it may be worth considering the effect of treatment, if any . With this in mind, diagnosis and differential diagnoses are reviewed, and with regard to treatment distinctions are made between neonatal conjunctivitis (gonococcus, chlamydia, virus), conjunctivitis in children (symptoms of upper respiratory tract infections), and conjunctivitis in adults and the elderly (dry eyes, epiphora in ectropion).
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