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 coastl