J Anim Sci, 1992 Oct, 70(10), 3055 - 65 Effect of long- or short-term feeding of alpha-tocopheryl acetate to Holstein and crossbred beef steers on performance, carcass characteristics, and beef color stability; Arnold RN et al.; Three experiments were conducted to examine the effects of vitamin E supplementation on feedlot cattle . Vitamin E supplementation did not affect feedlot performance or carcass characteristics of cattle fed a high-concentrate diet (P greater than .1) . The major finding was the effectiveness of vitamin E in extending the color stability of displayed beef (P less than .01) . Color stability during display of longissimus lumborum steaks from cattle supplemented with 300 IU/d for 266 d, 1,140 IU/d for 67 d, or 1,200 IU/d for 38 d was extended by 2.5 to 4.8 d . Gluteus medius steaks had an extended color display life of 1.6 to 3.8 d . The accumulation of lipid oxidation products, but not aerobic microbes, associated with displayed longissimus lumborum was suppressed for muscle from vitamin E-supplemented steers . Taste panelists detected no difference among longissimus lumborum steaks from control and vitamin E-supplemented steers but found (P less than .01) steaks aged for 21 d to be more tender than steaks aged for 7 d . Supplementing cattle with vitamin E should reduce economic losses associated with discolored beef during retail display. CLAO J, 1992 Oct, 18(4), 240 - 4 Microbial contamination of hydrophilic contact lenses . Part II: Quantitation of microbes after patient handling and after aseptic removal from the eye; Mowrey-McKee MF et al.; We cultured Soflens (polymacon) contact lenses to determine the number of microorganisms present after patient handling and the number present after patient handling and 5 hours of lens wear . Twenty adapted contact lens patients were each dispensed two pairs of new sterile Soflens contact lenses . Both pairs of lenses were handled by the patients during a single experimental session . Prior to handling lenses, patients were instructed to wash their hands with soap and water, rinse with tap water, and dry with a paper towel . One pair (handled-only) was cultured immediately after handling . The second pair (handled-and-worn) was placed on the eyes by the patient, removed aseptically following 5 hours of wear, and then cultured . All 40 handled-only lenses had viable microorganisms associated with them (mean: 653 colony forming units (CFU) per lens) . Bacteria were isolated on all of these lenses and fungi were detected on six . The mean bacterial count for the handled-and-worn lenses was 30 CFU/lens; fewer than half exhibited handled-and-worn lenses was 30 CFU/lens; fewer than half exhibited bacteria and no fungi were isolated . These data suggest that patient handling is a highly significant source of microbial contamination of hydrophilic contact lenses . Also, the number of microorganisms on lenses is significantly reduced (P < 0.0001) after the lens has been worn on the eye. J Periodontol, 1992 Oct, 63(10), 821 - 4 Efficacy of preprocedural rinsing with an antiseptic in reducing viable bacteria in dental aerosols; Fine DH et al.; This double-blind, controlled, cross-over, clinical study evaluated the effect of preprocedural rinsing with an antiseptic mouthrinse on the level of recoverable viable bacteria in an aerosol generated during a typical dental procedure . Eighteen subjects participated . Following 24 hours of abstention from all oral hygiene procedures, subjects received a 10-minute ultrasonic scaling of a randomly selected one-half of their mouth which served as the unrinsed control . They were then randomly assigned either antiseptic mouthwash or a control rinse and rinsed with 20 ml for 30 seconds, after which the remaining half mouth (experimental side) was scaled ultrasonically for 10 minutes . During each 10-minute scaling period aerosolized bacteria were collected on a sterile filter using a modified vacuum air-sampling device . Microbes captured on the sterile filter were quantitated by overlaying the filters onto trypticase soy agar, incubating the filters aerobically at 37 degrees C for 24 to 72 hours, and counting the resulting colony forming units (CFU) . Preliminary experiments had confirmed that neither the collection method nor residual antiseptic mouthwash in the aerosol adversely affected the number of viable bacteria recovered from the filter . Rinsing with the antiseptic mouthwash produced a 94.1% reduction in recoverable CFUs compared to the non-rinsed control, while the control rinse produced a 33.9% reduction . The difference between the mouthwash and control was statistically significant (P < .001) . This study indicates that preprocedural rinsing with an antiseptic mouthwash can significantly reduce the microbial content of aerosols generated during ultrasonic scaling and may have potential in-office use as part of an infection control regimen. Eur J Immunol, 1992 Oct, 22(10), 2555 - 63 Genetics of nonspecific immunity: I . Bidirectional selective breeding of lines of mice endowed with maximal or minimal inflammatory responsiveness; Ibanez OM et al.; The genetic regulation of acute inflammatory reaction (AIR) was studied by the method of bidirectional selective breeding, used to produce a line of mice giving the maximal and a line of mice giving the minimal inflammatory reaction (AIR max and AIR min, respectively) . The AIR was triggered by subcutaneous injection of a neutral substrate (suspension of polyacrylamide microbeads), and measured by the leukocyte and serum protein accumulation in the exudate . The two parameters are positively correlated and present a normal frequency distribution . The highly genetically heterogeneous foundation population was produced by the equipoised intercrossing of eight inbred strains of mice, and selective breeding carried out by assortative matings of extreme phenotypes . The response to selection in 11 consecutive generations was highly asymmetrical: a marked AIR increase in the AIR max and no change in the AIR min line occurred . The mean value of realized heritability in the AIR max line was 0.26 and 0.18 for cell and protein concentrations, respectively . The response to selection must have resulted from the interaction of seven to nine independent gene loci endowed with additive effects . The lack of response to selection of the AIR min line is discussed . The large inter-line difference opens new possibilities for studying the biochemistry and molecular genetics of inflammation, and also for investigating the beneficial or detrimental effect of inflammatory responses. J Cell Sci, 1992 Sep, 103 ( Pt 1), 39 - 51 Examination of transcellular membrane protein polarity of bovine aortic endothelial cells in vitro using the cationic colloidal silica microbead membrane-isolation procedure; Stolz DB et al.; In this report we describe a rapid, high-yield protocol for the isolation of apical (AP) and basolateral (BL) plasma membrane domains from monolayers of bovine aortic endothelial cells (BAECs) grown on tissue culture dishes as well as microcarrier beads . Using a modified cationic colloidal silica microbead membrane-isolation procedure, which deposits a uniform silica-polyacrylate pellicle over the entire AP membrane surface, a 4- to 9.6-fold relative enrichment of AP membrane and a 3.55- to 3.67-fold relative enrichment of BL membrane was obtained when the isolated domains were examined for silica and Na+/K(+)-ATPase, respectively . Immunoblotting of the isolated membrane domains displayed the presence of angiotensin-converting enzyme (ACE) exclusively in the AP domain and collagen receptors (CRs) highly enriched in the BL membrane domain when monolayers were grown on a gelatin substratum. J Dent Hyg, 1992 Sep, 66(7), 314 - 8 Aerosol generation by two ultrasonic scalers and one sonic scaler . A comparative study; Gross KB et al.; The purposes of this study were to compare the amount of aerosols generated from ultrasonic and sonic scalers and to measure the potential depth of respiratory tract penetration . Forty subjects were randomly assigned to receive instrumentation with the magnetostrictive, piezoelectric, or air turbine scaler . The Anderson Air Sampler collected total baseline airborne microbes for 20 minutes prior to treatment and for 20 minutes during instrumentation . This cascade impactor system measures the degree of microbial penetration in a simulated respiratory system . Blood agar plates from the sampler were incubated for 24 hours at 37 degrees C . Colony forming units per cubic foot of air (CFUs/cu . ft.) were enumerated by one blind examiner using a Lab Line Colony Counter . Data for total microbial CFUs/cu.ft . and CFUs/cu.ft . by sampler level were analyzed on the log-transformed data using ANCOVA . Baseline values of airborne bacteria served as the covariate . Results showed no significant difference in mean combined total CFUs/cu.ft . for the magnetostrictive, piezoelectric, or air turbine sonic scalers . The magnetostrictive scaler generated the lowest CFUs/cu.ft . at the deepest level of penetration; however, no significant difference in level of penetration was found among the three scalers. Mol Plant Microbe Interact, 1992 Sep-Oct, 5(5), 397 - 404 Resistance to tobacco mosaic virus induced by the 54-kDa gene sequence requires expression of the 54-kDa protein; Carr JP et al.; Tobacco plants transformed with the sequence encoding the 54-kDa putative replicase protein of tobacco mosaic virus were resistant to systemic virus disease (D . B . Golemboski, G . P . Lomonossoff, and M . Zaitlin, Proc . Natl . Acad . Sci . USA 87:6311-6315, 1990) . Resistance was due to a marked suppression of virus replication at the site of inoculation (J . P . Carr and M . Zaitlin, Mol . Plant-Microbe Interact . 4:579-585, 1991) . Although RNA transcripts encoding the 54-kDa protein were present in resistant plants, the 54-kDa protein itself was not observed in vivo . We wished to assess the relative importance of the 54-kDa protein versus its RNA in mediating resistance . Further attempts to detect the 54-kDa protein in plant tissues were unsuccessful; therefore, an indirect approach was taken using a protoplast-based transient gene expression system . Electroporation of protoplasts with plasmids capable of expressing the wild-type 54-kDa protein gene sequence or a mutant lacking the first AUG initiation codon of the 54-kDa open reading frame and encoding a slightly truncated protein reduced virus replication in protoplasts . In contrast, a frameshift mutant that was capable of directing synthesis of a protein only 20% the size of the 54-kDa protein, did not produce resistance in protoplasts . These results show that expression of the 54-kDa protein gene sequence at the RNA level alone is insufficient for resistance, and they implicate the 54-kDa protein itself in mediating this resistance phenomenon. Curr Opin Immunol, 1992 Aug, 4(4), 442 - 8 Old microbes with new faces: molecular biology and the design of new vaccines; Connell N et al.; Rational approaches to the design of live attenuated bacterial and viral recombinant vaccine strains are leading to the manipulation of old vaccines and the generation of new ones . The two basic problems to be solved are attenuation of pathogenic strains, and the stable expression of foreign antigens. FEBS Lett, 1992 Aug 10, 308(1), 94 - 6 A novel role for calcite in calcium homeostasis; Anderson S et al.; Calcium carbonate (CaCO3) minerals are known to be deposited in a wide array of different organisms, ranging from microbes to vertebrates {(1989) On Biomineralization, Oxford University Press, New York} . Calcite, aragonite and vaterite are the major crystalline structural polymorphs of CaCO3 associated with living systems, and participate in a variety of biological functions {(1989) Biomineralization: Chemical and Biochemical Perspectives, VCH Publishers, Weinham, Germany; (1991) Advances in Inorganic Chemistry 36, 137-200} . Here we report on the ability of a soil bacterium to synthesize calcite in a calcium-stressed environment . The elaboration of this exocellular crystalline residue enables the organism to regulate its calcium content . The attainment of calcium homeostasis via the exocellular deposition of bacterial calcite with unique crystal habits is a novel biological phenomenon. Inflammation, 1992 Aug, 16(4), 383 - 92 Stimulus-dependent actin polymerization in bovine neutrophils; Bochsler PN et al.; Polymorphonuclear leukocytes (PMNs) are responsible for much of the first wave of leukocyte-mediated host defense against microbial pathogens . In order to migrate through the endothelium of vessel walls, undergo chemotaxis, and phagocytize microbes, PMNs must modulate their cytoskeletal elements and undergo change of cellular shape . We have used fluorescence flow cytometric analysis and cellular microscopic observations to demonstrate actin polymerization in bovine PMNs and to examine the kinetics of PMN actin polymerization utilizing different PMN stimuli . In addition, we compared temporal relationships between cellular shape and actin polymerization . Actin polymerization occurred rapidly, and the kinetics of actin polymerization were similar for each of the three PMN agonists used, ZAS (10%), PAF (10(-6) M), and rhC5a (10(-7) M) . Actin polymerization was near-maximal by 10 sec poststimulation (95.4% of maximal F-actin content attained by 10 sec poststimulation with ZAS stimulation), and reached peak values by 30 sec . The maximal increase in F-actin content of agonist-stimulated cells as compared to resting cells was 2.8-fold with ZAS; 2.3-fold with PAF; and 2.3-fold with rhC5a . PMN shape change (pseudopodia, membrane ruffles) was not as rapid, with only 22.4% of cells attaining visible membrane deformation by 10 sec and requiring 120 sec to reach peak shape-change values . After attaining peak values, the two events also differed . Whereas the percent of shape-changed PMNs remained plateaued up to 5 min poststimulation, the F-actin content gradually decreased after 30 sec, approaching F-actin values of unstimulated PMNs. J Acoust Soc Am, 1992 Aug, 92(2 Pt 1), 688 - 700 Inferring articulation and recognizing gestures from acoustics with a neural network trained on x-ray microbeam data; Papcun G et al.; This paper describes a method for inferring articulatory parameters from acoustics with a neural network trained on paired acoustic and articulatory data . An x-ray microbeam recorded the vertical movements of the lower lip, tongue tip, and tongue dorsum of three speakers saying the English stop consonants in repeated Ce syllables . A neural network was then trained to map from simultaneously recorded acoustic data to the articulatory data . To evaluate learning, acoustics from the training set were passed through the neural network . To evaluate generalization, acoustics from speakers or consonants excluded from the training set were passed through the network . The articulatory trajectories thus inferred were a good fit to the actual movements in both the learning and generalization conditions, as judged by root-mean-square error and correlation . Inferred trajectories were also matched to templates of lower lip, tongue tip, and tongue dorsum release gestures extracted from the original data . This technique correctly recognized from 94.4% to 98.9% of all gestures in the learning and cross-speaker generalization conditions, and 75% of gestures underlying consonants excluded from the training set . In addition, greater regularity was observed for movements of articulators that were critical in the formation of each consonant. J Am Vet Med Assoc, 1992 Aug 1, 201(3), 478 - 82 Clinical and laboratory findings associated with actual or suspected azoospermia in dogs: 18 cases (1979-1990); Olson PN et al.; Eighteen dogs were evaluated for azoospermia, 8 of which had sired pups . On the basis of history, physical examination, and various laboratory evaluations, the cause and site of azoospermia varied . Two dogs that had never sired pups had likely been azoospermic from puberty (congenital azoospermia) . Two dogs were azoospermic as a result of tumors (Sertoli cell tumor and malignant astrocytoma of the pituitary gland) . Deposits of IgG were observed in testicular biopsy samples, which suggested an auto-immune cause for azoospermia in 5 dogs . One of the 5 dogs with IgG deposits in testicular tissues also had evidence of immune-mediated thyroiditis . Culturing of microbes in the semen was not helpful in determining potential causes of azoospermia, and results did not correlate with organisms isolated from testicular biopsy samples or with the finding of inflammation in biopsy samples . Because 6 dogs had relatives with histories of reproductive dysfunction, inbreeding also must be considered when evaluating dogs for azoospermia. J Cell Biol, 1992 Aug, 118(4), 865 - 75 Tubulin protofilaments and kinesin-dependent motility; Kamimura S et al.; Microtubules are built of tubulin subunits assembled into hollow cylinders which consist of parallel protofilaments . Thus, motor molecules interacting with a microtubule could do so either with one or several tubulin subunits . This makes it difficult to determine the structural requirements for the interaction . One way to approach the problem is to alter the surface lattice . This can be done in several ways . Proto-filaments can be exposed on their inside (C-tubules or "sheets"), they can be made antiparallel (zinc sheets), or they can be rolled up (duplex tubules) . We have exploited this polymorphism to study how the motor protein kinesin attached to a glass surface interacts and moves the various tubulin assemblies . Microtubules glide over the surface along straight paths and with uniform velocities . In the case of C-tubules, approximately 40% glide similarly to microtubules, but a major fraction do not glide at all . This indicates (a) that a full cylindrical closure is not necessary for movement, and (b) that the inside surface of microtubules does not support gliding . With zinc sheets, up to 70% of the polymers move, but the movement is discontinuous, has a reduced speed, and follows along a curved path . Since zinc sheets have protofilaments alternating in orientation and polarity, this result suggests that in principle a single protofilament can produce movement, even when its neighbors cannot . Duplex microtubules do not move because they are covered with protofilaments coiled inside out, thus preventing the interaction with kinesin . The data can be explained by assuming that the outside of one protofilament represents the minimal track for kinesin, but smooth gliding requires several parallel protofilaments . Finally, we followed the motion of kinesin-coated microbeads on sea-urchin sperm flagella, from the flagellar outer doublet microtubules to the singlet microtubule tips extending from the A-tubules . No change in behavior was detected during the transition . This indicates that even if these microtubules differ in surface lattice, this does not affect the motility. Voen Med Zh, 1992 Aug, (8), 20 - 3 {A comparative study of charcoal sorbents for local wound treatment}; Adamian AA et al.; The laboratory and medico-biological findings made it possible to choose the most suitable carbonaceous sorbents for local management of wounds, basing on the fact that every carbonaceous material has strictly selected capacity for protein and microbe sorption, and also can be the carrier of the water-miscible drugs . The "Dnieper-MN" carbonaceous fabric and "Aktilen" fibrous sorbent have the best features for sorption . It is possible to use these materials for the construction of bandages to treat wounds and burns which have little exudation . "Karpema" and "AUTM-2" sorbents, which are characterized by the low degree of activation, can be used for the manufacturing of surgical pads . "Karpema" being strong and having a less fiber structure, has a good electrical conductivity and, thus, can be used as medical electrodes. Gesundheitswesen, 1992 Aug, 54(8), 420 - 2 {Sanitation measures in evidence of Legionella}; Bosenberg H et al.; Appearance of Legionella disease generally is underrated . Conclusions as to disease frequency from Legionella antibody investigations are not reliable . The number of microbes necessary to produce illness depends from individual preliminary conditions . In special areas of the hospital the rate of diseases caused by Legionella can be reduced by protection . In bathing areas generally the hitherto existing regulations of the Federal Health Authority are sufficient: Continual warm water temperature increases from 60 degrees C, chlorination if necessary, regular stepwise controls . Insufficient results are produced by intermittent temperature increases . Filters that are impermeable for microbes appear uneconomical for bathing areas. Biotechniques, 1992 Aug, 13(2), 276 - 81 Analysis of mixed human/microbial DNA samples: a validation study of two PCR AMP-FLP typing methods; Lienert K et al.; The reliability of the PCR technique used to type two human variable number tandem repeats, that is, 3' to apolipoprotein B gene and locus D17S30, was examined using DNA samples of mixed human and microbial origin . Mixtures of human and microbial DNA were amplified, choosing microbes found commonly in the vagina . Total inhibition of human amplification and/or "drop-out" of the larger amplification fragment length polymorphism allele was observed at both loci in the presence of DNA from some vaginal micro-flora. Biochim Biophys Acta, 1992 Jul 31, 1122(2), 143 - 6 Lipases from different sources vary widely in dependence of catalytic activity on water activity; Valivety RH et al.; We have measured the rates of esterification in hexane catalysed by suspended immobilised lipases (triacylglycerol acylhydrolase, EC 3.1.1.3), with pre-equilibration to known thermodynamic water activity (a(w)) . There were important differences between the enzymes from five different microbes in their retention of activity at low a(w) . That from Rhizomucor miehei showed over 40% maximal activity at an a(w) of 0.12, and that from Rhizopus niveus was also fairly active at low a(w) . Lipases from other sources required higher a(w) values to show good activity, increasing in the sequence Humicola sp., Candida rugosa and Pseudomonas cepacia . The behaviour was generally similar to two very different support materials, anion-exchange resin and macroporous polypropylene . Comparison of the sequences of the homologous enzymes from Rh . miehei, Rh . niveus and Humicola sp . suggests that changes in charged residues in the 'hinge and lid' region of the structure may be significant in low a(w) tolerance. Experientia, 1992 Jul 15, 48(7), 640 - 3 The cellular immune response to heat shock proteins; Kaufmann SH; T lymphocytes, which are central to almost every immune response, frequently recognize microbial hsp60 . Such cells could provide an early defense mechanism against pathogenic microbes . However, T cells also recognize epitopes of hsp60 shared by microbe and host . Not only conventional alpha/beta T cells respond to hsp60; gamma/delta T cells do so, as well . In fact, certain gamma/delta T cells seem to have a particular preference for this molecule . Recognition of stressed host cells expressing hsp60 could facilitate the scavenger function of the T cell system . On the other hand, such recognition could be involved in autoimmune disease. Lab Anim, 1992 Jul, 26(3), 206 - 10 The effects of intracage ventilation on microenvironmental conditions in filter-top cages; Lipman NS et al.; Filter-top cages, while effective in reducing cross contamination by particulate material including microbes, can also cause accumulation of the waste gases carbon dioxide and ammonia as well as increased intracage relative humidity . A prototype system which provided each cage with 23 air changes per hour through a nozzle inserted in the filter lid was evaluated . The ventilated cageing system was effective in reducing intracage carbon dioxide, ammonia and relative humidity levels . Mean weekly carbon dioxide levels were 2900 ppm lower, ammonia levels 240 ppm lower and intracage relative humidity levels 8% lower in the ventilated cages than in unventilated controls. J Biomed Mater Res, 1992 Jul, 26(7), 967 - 77 Agarose for a bioartificial pancreas; Iwata H et al.; Islets were encapsulated into 5% concentration agarose microbeads . The effect of microencapsulation on islet allograft survivals was determined using a streptozotocin-induced diabetic (STZ) mouse and a nonobese diabetic (NOD) mouse as recipients . All five STZ BALB/c mice receiving microencapsulated islets (C57BL/6) maintained normoglycemia indefinitely . When NOD mice were used as recipients of the bioartificial pancreas, four of five grafts (islets from C3H/He) functioned for more than 80 d . Two of five NOD mice maintained normoglycemia until animals were sacrificed at 102 and 192 postoperative d . Microbeads made of commercially available agarose can effectively prolong alloislets functioning in the STZ-diabetic mouse and even in the NOD mouse (animal model of human type I diabetes) without the use of any immunosuppressive drug. ASAIO J, 1992 Jul-Sep, 38(3), M386 - 9 Inflammatory reaction induced by agarose implants reduced by adding adrenal cells to the polymer; Cadic-Amadeuf CM et al.; Microencapsulation of adrenal cells is proposed for reducing the non-specific inflammatory reaction observed around polymer implants . This hypothesis was tested by comparing both host cellular reaction and the surrounding graft cell populations that appeared when either agarose embedded cells or empty agarose beads were implanted . The authors' results showed that the fibrotic material that surrounded the implanted empty agarose microbeads was not as severe when adrenal cells were present . Similarly, the T lymphocyte population surrounding the graft was considerably reduced, along with the percentage of CD4 and CD8 positive cell subpopulations . The activation macrophage marker IaD disappeared . The authors' results support the hypothesis that embedded adrenal cells may be a suitable solution for reducing early inflammatory events due to microcapsule implantation. Environ Health Perspect, 1992 Jul, 97, 171 - 5 Activity testing of alveolar macrophages and changes in surfactant phospholipids after irradiation in bronchoalveolar lavage: experimental and clinical data; Steinberg F et al.; This study presents results of bronchoalveolar lavage (BAL) after irradiation to the lungs in mice as well as clinical data . The number of BAL cells, mainly macrophages, lymphocytes, and granulocytes, changed in a time-dependent manner . The phagocytic activity of the macrophages measured as the phagocytosis of microbeads and measured as the esterase activity also showed a strong time-dependent increase during the acute phase up to 21 days after irradiation . The contents of surfactant phospholipids (SF) and sphingomyelin (SPH; as a parameter for cell death) were quantified by HPLC . Both were significantly changed between day 2 and 21 after irradiation . Three BALs of a patient with idiopathic interstitial pneumonitis, who had received an allogenic bone marrow graft after total body irradiation with 10 Gy, showed similar effects in the cellular and surfactant parameters . These data indicate that there are positive interactions between the number of different BAL cells, macrophage activity, and SF and SPH content in the preclinical model of the mouse as well as in the clinical situation after lung irradiation. Mikrobiol Zh, 1992 Jul-Aug, 54(4), 3 - 7 {The interaction of the causative agent of melioidosis with the host's alveolar macrophages}; Popov SF et al.; Studies were carried out on guinea pigs and albino rats, intranasally infected with P . pseudomallei C-141 . The cells of bronchovesicular exudate were obtained from animals 1, 4 and 24 hours after infection . Electron microscopy was applied to study the process of interaction of the agent and alveolar macrophages . Bacteria were shown to form a capsule which permitted avoiding phagocytosis, when entering the host respiratory system . Microbes that failed to form a capsule were absorbed by macrophages and enclosed in a phagosome . Then some bacteria were destroyed by the lysosomal enzymes, the other synthesized a capsule, which protected them against the effect of phagolysosome content . There were also such microbes which escaped from a phagosome prior to fusion with lysosomes and parasitized in phagocytic cytoplasma forming a capsule there . By the end of the first 24 hours of observation the intact encapsulated microbe species were found to prevail in the host cells. Ann Acad Med Singapore, 1992 Jul, 21(4), 571 - 5 Human ampullary co-cultures for blastocyst transfer in assisted reproduction; Bongso TA et al.; Although the assisted reproductive techniques (ART) have contributed significantly over the last decade in alleviating subfertility in the childless couple, the implantation and take-home baby rates have been stubbornly low . A major cause for such low success rates has been the reduced viability of replaced embryos perhaps induced by the suboptimal in vitro conditions used in ART laboratories . One approach to improving embryo viability is to provide the growing embryo with a simulated in vivo environment by replicating the conditions existing in the human fallopian tube in vitro . This requires either the maintenance of an intact fallopian tube in vitro or establishment and maintenance of tubal epithelial cell-lines which could be used as feeder layers for early embryonic growth . The concomitant growth of cells with embryos in vitro has been referred to as co-culture . This paper discusses the in vitro behaviour of human tubal epithelial cells, the fertilisation and growth of embryos in ampullary co-culture, the specificity of co-cultures, the mechanism of action of co-cultures and the methods of screening the human ampullary co-culture system for microbes . The pregnancy and implantation results on 50 patients enrolled for a co-culture clinical trial are presented and the future use of this system discussed. Biochem J, 1992 Jun 15, 284 ( Pt 3), 781 - 8 Cytochrome b-245 is a flavocytochrome containing FAD and the NADPH-binding site of the microbicidal oxidase of phagocytes; Segal AW et al.; The NADPH oxidase of phagocytic cells is important for the efficient killing and digestion of ingested microbes . A very unusual low-potential cytochrome b (b-245) is the only redox molecule to have been identified in this system . The FAD-containing flavoprotein that binds NADPH and transfers electrons to the cytochrome has eluded identification for three decades . We show here that the haem/FAD ratio in the membranes does not change significantly on activation of this oxidase, indicating that the FAD is present in the membranes from the outset and not recruited from the cytosol . The FAD content of membranes from cells of patients with X-linked chronic granulomatous disease (CGD) lacking the cytochrome b was roughly one-quarter of that in normal subjects and in autosomal recessive CGD patients lacking the cytosolic protein p47-phox . Similar low amounts of FAD were present in uninduced promyelocytic (HL60) cells, suggesting that the low amount of FAD in cells from X-CGD patients was probably unrelated to this oxidase system . Cytochrome b-245 appears to bind both the haem and FAD, in a molar ratio of 2:1 . The e.p.r . signal of the purified cytochrome was weak and had an asymmetric g(z) peak at g = 3.31 . The purified cytochrome could be partially reflavinated (about 20%) in the presence of lipid . Amino acid sequence homology was detected between the beta-subunit of this cytochrome b and the ferredoxin-NADP+ reductase (FNR) family of reductases in the putative NADPH- and FAD-binding sites . 32P-labelled 2-azido-NADP was used as a photoaffinity label for the NADPH-binding site . Labelling that was competed off with NADP was observed in the region of the beta-subunit of the cytochrome . No labelling was seen in this region in X-CGD in three subjects in whom this cytochrome was missing and in a third in whom it was present but bore a Pro-His transposition in the putative NADPH-binding site . These studies indicate that cytochrome b-245 is a flavocytochrome, the first described in higher eukaryotic cells, bearing the complete electron-transporting apparatus of the NADPH oxidase. Nature, 1992 Jun 11, 357(6378), 515 - 8 Novel form of growth cone motility involving site-directed actin filament assembly; Forscher P et al.; Regulation of cytoskeletal structure and motility by extracellular signals is essential for all directed forms of cell movement and underlies the developmental process of axonal guidance in neuronal growth cones . Interaction with polycationic microbeads can trigger morphogenic changes in neurons and muscle cells normally associated with formation of pre- and postsynaptic specializations . Furthermore, when various types of microscopic particles are applied to the lamellar surface of a neuronal growth cone or motile cell they often exhibit retrograde movement at rates of 1-6 microns min-1 (refs 3-6) . There is strong evidence that this form of particle movement results from translocation of membrane proteins associated with cortical F-actin networks, not from bulk retrograde lipid flow and may be a mechanism behind processes such as cell locomotion, growth cone migration and capping of cell-surface antigens . Here we report a new form of motility stimulated by polycationic bead interactions with the growth-cone membrane surface . Bead binding rapidly induces intracellular actin filament assembly, coincident with a production of force sufficient to drive bead movements . These extracellular bead movements resemble intracellular movements of bacterial parasites known to redirect host cell F-actin assembly for propulsion . Our results suggest that site-directed actin filament assembly may be a widespread cellular mechanism for generating force at membrane-cytoskeletal interfaces. Am J Physiol, 1992 Jun, 262(6 Pt 1), L708 - 12 Release of reactive oxygen species by guinea pig tracheal epithelial cells in vitro; Kinnula VL et al.; Regulatory and stimulatory mechanisms of H2O2 release from guinea pig tracheal epithelial cells were investigated . Cells in primary culture maintained in a previously described air-liquid interface system released H2O2 to the extracellular space only from the apical side of the cells . The rate of release was 0.044 +/- 0.003 nmol.min-1.mg protein-1 . H2O2 release could be stimulated significantly during a 30-min incubation period with phorbol myristate acetate (PMA) and platelet-activating factor (PAF) . A stimulatory effect of PAF was achieved at concentrations greater than 100 nM and with PMA at concentrations greater than 10 ng (16 nM) . When protein kinase C was inactivated with staurosporine, the responses to both PAF and PMA were abolished, whereas the cyclooxygenase inhibitor, indomethacin, did not affect H2O2 generation . When guinea pig tracheal epithelial cells were exposed to sublethal concentrations of extracellular H2O2 (30 microM), H2O2 was detoxified from both apical and basal sides, H2O2 removal being significantly more rapid from the apical side of the cells . These results suggest that tracheal epithelial cells can be stimulated to generate reactive oxygen species into the airway lumen and that this occurs in response to inflammatory mediators that act through protein kinase C . Luminal H2O2 release may have developed as a defense mechanism against microbes, and, similarly, luminal detoxification of H2O2 could represent an important mechanism of modulation of airway inflammation in response to oxidant stress. Fertil Steril, 1992 Jun, 57(6), 1225 - 9 Cyclic variation of major histocompatibility complex class II antigen expression in the human fallopian tube epithelium; Edelstam GA et al.; OBJECTIVE: To systematically investigate the expression of major histocompatibility complex (MHC) class II antigens (human leukocyte antigens, HLA-DR, -DP, and -DQ) on columnar epithelium in the fallopian tube during the menstrual cycle . STUDY DESIGN: Biopsies were collected from the fallopian tube during laparotomy sterilization and immunoperoxidase staining was performed . SETTINGS: Departments of Gynecology and Obstetrics and Clinical Immunology and Transfusion Medicine, Uppsala University, Uppsala, Sweden . PATIENTS: Twenty healthy fertile women undergoing sterilization at different times of the menstrual cycle . INTERVENTIONS: None . MAIN OUTCOME MEASURES: The staining of the columnar epithelium was judged on a 4-graded scale according to the distribution of class II antigens . RESULTS: A widespread preovulatory HLA-DR expression was observed almost completely lining the columnar epithelial cells including the luminal surface, whereas postovulatory the HLA-DR expression was withdrawn from the surface . The HLA-DP and -DQ antigens varied in a similar way, although not as pronounced . CONCLUSIONS: The MHC class II antigen variation in the fallopian tube epithelium seen in this study may indicate a hormonal regulation that could reflect variable need for local immunocompetence during the menstrual cycle: a preovulatory need for immunoreactivity against invading microbes and postovulatory an optimal survival of the foreign preimplantation embryo. J Oral Maxillofac Surg, 1992 May, 50(5), 477 - 82; discussion 482-3 Autologous facial fat transplantation: improved graft maintenance by microbead bioactivation; Eppley BL et al.; An alternative approach to the management of free fat transplantation resorption was evaluated in a rat facial model . Fat grafts obtained from the inguinal region were transferred to subcutaneous lateral facial sites in 20 animals . The grafts were mixed with either basic fibroblast growth factor (bFGF) alone or dextran beads that had been pretreated with bFGF . The grafts were then compared by weight and histology at 1 and 6 months postoperatively . Although graft weights were nearly comparable at 1 month, substantial differences were seen at 6 months, with the bead-containing grafts exhibiting near complete weight maintenance and better overall graft form . Histologically, the bead-containing grafts had extensive intercellular collagen formation and a heterogeneity of adipocyte cell sizes, particularly after 1 month . These findings suggest that the addition of cell-specific bioactive peptides that affect either the preadipocyte cell line and/or the fibroblastic components of the recipient site improve postoperative fat graft weight maintenance . Delivery of the biochemical agent appears to require a carrier system to exert its effects. J Can Dent Assoc, 1992 May, 58(5), 377 - 8, 382-6 Concerns regarding infection control recommendations for dental practice; Hardie J; It goes without saying that the members of any professional group are more likely to modify their behavior if they are provided with logical, rational reasons to enact the suggested change . In the mid 1980s, health care providers, including dental personnel, were advised to adopt universal precautions and to alter their infection control habits with minimal justification, apart from the general unease and paranoia surrounding AIDS . Therefore, it is understandable that some practitioners would react with scepticism to the idea that their traditional infection control techniques were less than adequate, while others would overwhelmingly embrace the new recommendations in the misguided belief that personal, patient, staff and family safety would be enhanced . This predictable confusion is epitomized by the dentist who "sterilizes" extraction forceps by immersing them in alcohol for 10 minutes, versus the dentist who wears gloves, mask and disposable gown to conduct a recall examination . And if dentists are perplexed, it is clear that their staffs are equally, if not more confused, since they are exposed to the exaggerated claims and counter claims of sales agents . The microbes encountered in dental practise, apart from the hepatitis B virus, pose no significant risk to dental personnel or their patients, and the danger of hepatitis B transmission is reduced most effectively by vaccination . In reality, the genesis of dentistry's current emphasis on infection control resides entirely with HIV disease . But there is no credible clinical evidence to suggest that HIV infection is transmitted via dental treatment . Indeed, it may be theorized that for such a transmission to occur, the blood stream of the susceptible recipient would have to be invaded directly by a pathogenic inoculum of the virus--an unlikely event in the normal practise of dentistry . Under such circumstances, infection control practises should ignore the danger of HIV transmission, but concentrate on: Sterilization of all surgical and invasive instruments to protect patients from potential cross-infection . All dental staff receiving hepatitis B vaccinations . Dental staff wearing gloves, especially while performing intraoral procedures with blood release, and handling used instruments, to protect them from direct contact with potential pathogens . Working in a clean environment, in which blood spills and splatters are removed mainly for esthetic reasons . Such measures reflect the actual potential for disease transmission, as it exists in dentistry . They are justified and economical, and will be implemented by concerned but knowledgeable dental staff. Br J Ind Med, 1992 May, 49(5), 332 - 6 Class specific antibodies in serodiagnosis of farmer's lung; Ojanen T; The aim of the present study was to determine which microbes and which immunoglobulin (Ig) classes should be included in tests to discriminate between patients with farmer's lung and reference persons . The sera of a group of farmer's lung patients and their spouses were measured for IgG, IgA, IgM, and IgE antibodies against a panel of farmer's lung microbes . The concentrations of IgG, IgA, and IgE antibodies were higher in patients compared with their spouses . The patients were generally positive for antibodies of several Ig classes whereas the spouses had only either IgG or IgA antibodies . A test comprising the determinations of IgG antibodies against T vulgaris and IgA antibodies against A fumigatus would correctly group 94% of the cases in the Finnish farming population . The selection of microbes for other environments needs to be determined locally. Antibiot Khimioter, 1992 May, 37(5), 30 - 1 {Antibiotics of the aminoglycoside group (gentamicin, sisomicin and amikacin) in the prevention and treatment of experimental plague}; Shcherbaniuk AI et al.; Activity of aminoglycosides such as gentamicin, sisomicin and amikacin against plague microbe strains of natural origin was studied in vitro . It was also studied in prophylaxis and treatment of experimental plague infection in albino mice . The MAC of gentamicin and sisomicin for 50 strains of the plague microbe was 0.2-1.6 micrograms/ml . For the majority of the strains it was 0.4 micrograms/ml . The amikacin MICs were 0.4-3.2 and 0.8 micrograms/ml, respectively . High efficacy of gentamicin, sisomicin and amikacin was shown in prophylaxis and treatment of experimental plague infection in albino mice . The optimal doses of the antibiotics were determined . Under definite conditions such as the use of short-term regimens and higher intervals, advantages of sisomicin over gentamicin and amikacin in prophylaxis of experimental plague infection were observed. J Immunol Methods, 1992 Apr 27, 149(1), 1 - 10 A critical evaluation of the magnetic cell sorter and its use in the positive and negative selection of CD45RO+ cells; Manyonda IT et al.; In this paper, we report on our year-long experience with the magnetic cell sorter (MACS), and present a critical evaluation of its pitfalls and benefits . Satisfactory separation of lymphocytes into subsets with preservation of function can be achieved, but there are several drawbacks: in comparison with Dynal beads, MACS results in a higher cell loss due to the increased number of separation steps and requires depletion of plastic-adherent cells as these will engulf microbeads and contaminate the enriched fraction, and is more expensive . The advantage of MACS over Dynal beads is that the microbeads are biodegradable and do not interfere with proliferation assays: both the depleted and enriched fractions of cells can therefore be used in culture immediately following separation . We used MACS for the positive and negative selection of CD45RO cells: the enriched fraction was of high purity (greater than 98%), but a depleted fraction of greater than 90% purity could not be obtained even after running the same sample over three separating columns . Dynabeads, on the other hand, achieved 98% pure CD45RO-depleted fractions after three separation runs. CLAO J, 1992 Apr, 18(2), 87 - 91 Microbial contamination of hydrophilic contact lenses . Part I: Quantitation of microbes on patient worn-and-handled lenses; Mowrey-McKee MF et al.; We cultured Soflens (polymacon) contact lenses to determine the number of microorganisms present following normal patient wear and handling just prior to disinfection . Total protein deposited was determined for the companion lens from each patient . A random population of 109 adapted soft contact lens patients participated in the study . Some patients participated more than once, resulting in a total of 196 lenses being cultured and 195 lenses analyzed for total protein . The left lens was cultured immediately . The right lens was extracted at 70 degrees C in sodium hydroxide, and the total protein in the extract determined using a modified Lowry protein assay . The mean protein deposition per lens was 3.4 micrograms (median 2 micrograms per lens; range less than 1 to 78 micrograms/lens) . Microorganisms were cultured from 95% of the lenses . The mean bacterial count (in colony forming units per lens) was 2,482 (median: 123; range less than 3 to 150,000) . Fungal contamination was found on 11% of the lenses at very low levels (3-9 yeast/lens and 3-18 mold/lens) . Statistical analysis found no significant relationship between bacterial bioburden and any of the study parameters, including total protein, lens age, or subjective evaluation of lens cleanliness. Mol Microbiol, 1992 Apr, 6(7), 825 - 31 The suicidal DNA repair methyltransferases of microbes; Samson L; Virtually every organism so far tested has been found to possess an extremely efficient DNA repair mechanism to ensure that certain alkylated oxygens do not accumulate in the genome . The repair is executed by DNA methyltransferases (MTases) which repair DNA O6-methylguanine (O6MeG), O4-methylthymine (O4MeT) and methylphosphotriesters (MePT) . The mechanism is rather extravagant because an entire protein molecule is expended for the repair of just one, or sometimes two, O-alkyl DNA adduct(s) . Cells profit from such an expensive transaction by earning protection against death and mutation by alkylating agents . This review considers the structure, function and biological roles of a number of well-characterized microbial DNA repair MTases. J Cell Biol, 1992 Apr, 117(1), 105 - 20 Differential behavior of photoactivated microtubules in growing axons of mouse and frog neurons; Okabe S et al.; To characterize the behavior of axonal microtubules in vivo, we analyzed the movement of tubulin labeled with caged fluorescein after activation to be fluorescent by irradiation of 365-nm light . When mouse sensory neurons were microinjected with caged fluorescein-labeled tubulin and then a narrow region of the axon was illuminated with a 365-nm microbeam, photoactivated tubulin was stationary regardless of the position of photoactivation . We next introduced caged fluorescein-labeled tubulin into Xenopus embryos and nerve cells isolated from injected embryos were analyzed by photoactivation . In this case, movement of the photoactivated zone toward the axon tip was frequently observed . The photoactivated microtubule segments in the Xenopus axon moved out from their initial position without significant spreading, suggesting that fluorescent microtubules are not sliding as individual filaments, but rather translocating en bloc . Since these observations raised the possibility that the mechanism of nerve growth might differ between two types of neurons, we further characterized the movement of another component of the axon structure, the plasma membrane . Analysis of the position of polystyrene beads adhering to the neurites of Xenopus neurons revealed anterograde movement of the beads at the rate similar to the rate of microtubule movement . In contrast, no movement of the beads relative to the cell body was observed in mouse sensory neurons . These results suggest that the mode of translocation of cytoskeletal polymers and some components of the axon surface differ between two neuron types and that most microtubules are stationary within the axon of mammalian neurons where the surface-related motility of the axon is not observed. Khirurgiia (Mosk), 1992 Apr, (4), 77 - 80 {The modelling of diffuse peritonitis}; Ashurmetov RI et al.; Peritonitis is still an urgent problem and its solution is determined to a great measure by purposeful studies in the field of pathogenesis and treatment . The creation of adequate models of peritonitis acquires particular significance . Eighty-four animals (dogs) were used to create the model . It is shown in the work that a standard model of generalized peritonitis can be produced by two injections of a polymicrobial suspension consisting of the most commonly encountered peritoneal microbes E . coli and B . fragilis . The suspension is injected into different levels of the abdominal cavity . With this method of administration of the polymicrobial suspension the model was reproduced in 90% of cases . Thus, the developed model resembles human peritonitis in the clinical picture and the laboratory and morphological values. J Autoimmun, 1992 Apr, 5 Suppl A, 11 - 26 The forces driving autoimmune disease; Roitt IM et al.; There are two classes of autoimmune disease, organ-specific and non-organ specific or systemic . That cells producing autoantibodies are selected by antigen is strongly suggested by the presence of mutations and high affinity antibody . T-cells are pivotal in all forms of autoimmunity as evidenced by the therapeutic benefit of anti-T-cell monoclonals such as anti-CD4, and the frequent development of high affinity IgG autoantibodies . The production of anergic T-cells by the use of non-depleting anti-CD4 in the presence of antigen is discussed with particular reference to its potential for immunological intervention in autoimmune disease . It is possible to identify T-cell epitopes in organ-specific autoimmunity using pathogenic T-cell clones or hybridomas to identify the peptide sequences which are reactive . Antigen-specific therapy may ultimately be based on such peptide epitopes . The specificity of the T-cells in systemic autoimmunity is still obscure, but there is some evidence that reactivity with certain germ-line idiotypes can lead to the development of systemic autoimmunity . The possibility of stimulating B-cells specific for auto-antigens such as DNA becomes feasible if a complex of antibody and DNA is taken up by these specific B-cells and processed idiotype is presented to T-helpers specific for those idiotype epitopes . Evidence is presented that there may be pre-existing defects in the target organ in certain organ-specific disorders, and the evidence for a glycosylation defect in the IgG in patients with rheumatoid arthritis is explored . It is noted that the spouses of probands with rheumatoid arthritis is explored . It is noted that the spouses of probands with rheumatoid arthritis also tend to have this glycosylation defect and this raises the possibility of an effect due to an environmental factor, such as a microbial infection . Molecular mimicry of autoantigens by microbes can stimulate autoreactive cells by their cross-reactivity . It is emphasized that cross-reaction which gives rise to the priming of autoreactive T-cells could give rise to the establishment of a chronic autoimmune state . In animals with normal regulatory immune systems, such induced autoimmunity is ultimately corrected and it is only in animals where there are defects in regulation, that autoimmunity persists . Thus, there are many factors giving rise to autoimmunity, and the diseases are rightly regarded as multifactorial in origin. Eur J Biochem, 1992 Mar 15, 204(3), 991 - 1001 Molecular cloning and regulatory analysis of the cuticle-degrading-protease structural gene from the entomopathogenic fungus Metarhizium anisopliae; St Leger RJ et al.; The proteinaceous insect cuticle is an effective barrier against most microbes, but entomopathogenic fungi can breach it using extracellular proteases . We report here the isolation and characterization of a cDNA clone of the cuticle-degrading protease (Pr1) of Metarhizium anisopliae . The cDNA sequence revealed that Pr1 is synthesized as a large precursor (40.3 kDa) containing a signal peptide, a propeptide and the mature protein predicted to have a molecular mass of 28.6 kDa . The primary structure of Pr1 has extensive similarity with enzymes of the subtilisin subclass of serine endopeptidases and the serine, histidine and aspartate components of the active site in subtilisins are preserved . Proteinase K demonstrated the closest sequence similarity to Pr1 (61%) but Pr1 was twofold more effective than proteinase K at degrading isolated cuticles of Manduca sexta and 33-fold more effective at degrading structural proteins bound to the cuticle by covalent bonds . We postulate that the additional positively charged residues on the surface of the Pr1 molecule, as determined using proteinase K, may facilitate electrostatic binding to cuticle proteins which is a prerequisite for activity . Northern-blot analysis of RNA and nuclear run-on assays demonstrated transcriptional control of the expression of Pr1 during nutrient deprivation and during the formation of infection structures . Southern-blot analysis demonstrated that genes with significant homologies to Metarhizium Pr1 were present in the entomopathogens Aspergillus flavus and Verticillium lecanii but not Zoophthora (= Erynia) radicans. Scanning Microsc, 1992 Mar, 6(1), 167 - 74; discussion 174-5 Microbeam studies of the sensitivity of structures within living cells; Braby LA; Determining the biological effects of low doses of radiation with high linear energy transfer (LET) is complicated by the stochastic nature of charged-particle interactions . Populations of cells exposed to very low radiation doses contain a few cells which have been hit by a charged particle, while the majority of the cells receive no radiation damage . At somewhat higher doses, a few cells receive two or more events . Because the effects of damage produced by separate events can interact in the cell, we have had to make assumptions about the nature of these interactions in order to interpret the results of the experiments . Many of those assumptions can be tested if we can be sure of the number of charged-particle events which occur in individual cells, and correlate this number with the biological effect . We have developed a special irradiation facility at Pacific Northwest Laboratory (PNL) to control the actual number of charged particle tracks that pass through cell nuclei . The beam from a 2 MeV tandem accelerator is collimated to approximately 5 microns . Cells, grown in special dishes with 1.5 microns thick plastic bottoms, are positioned so that the desired portion of the cell aligns with the collimator . A shutter in the beam line is opened and closed after the desired number of particle tracks has been counted . This approach can be used to investigate the effects of the interaction between irradiated and unirradiated cells in an organized system, as well as to study the effects of spatial and temporal distribution of radiation damage within single cells.(ABSTRACT TRUNCATED AT 250 WORDS) J Burn Care Rehabil, 1992 Mar-Apr, 13(2 Pt 2), 287 - 92 Natural resistance to infection: leukocyte functions; Benhaim P et al.; The nonspecific immune system has an extraordinarily important role in prevention of and response to wound infection . In the presence of injury or contamination, leukocytes adhere to local endothelial cells in response to complement signals through an integrin mechanism; they then migrate to the affected tissue site to locate, ingest, and kill microbes . Clinically, the major weaknesses in this system are impairment of cell recruitment and depression of microbial killing mechanisms; intracellular killing is the most problematic step in the process . Provided that leukocytes are able to migrate and ingest bacteria, nonoxidative killing proceeds in a normal fashion . Oxidative killing, however, can be profoundly impaired by poor blood perfusion and oxygenation . Low oxygen tension is a common clinical problem in surgical sites and wounds . There are a number of ways in which oxidative killing can be supported and facilitated in the clinical arena in a manner that is likely to enhance the effects of antibiotics in the treatment and prevention of infection. Nippon Geka Hokan, 1992 Mar 1, 61(2), 190 - 8 Adenomyomatosis of the gallbladder: a clinical survey of 30 surgically treated patients; Kasahara Y et al.; Thirty patients with adenomyomatosis of the gallbladder (AMG) were operated on between January 1983 and June 1990 . They were made up 3.3% of patients who underwent cholecystectomy during the same interval . Of the 30 patients, ages ranged from 22 to 77 years (mean 52.3 years) and the male-to-female ratio was 8:7 . Among the macroscopic types, 10 cases of generalized, 12 of segmental (S) and 8 of fundal (F) were noted, and the size of the affected portion in type S (0.8 +/- 0.2 cm, mean +/- SD) was significantly thinner than in other two types (p less than 0.05) . Although the main symptom was abdominal pain, the majority of patients with type F had no complaints . Twenty patients (27%) were accompanied by gallstones including cholesterol stones in 60% of cases, and all six cases showing microbes in the bile had gallstones . Only six patients were diagnosed as AMG by preoperative imaging techniques . Other diagnoses comprised 15 of chronic cholecystitis and 3 of suspected gallbladder carcinoma . To identify the expanded Rokitansky-Aschoff sinuses, endoscopic retrograde cholangiography and/or ultrasonography of the abdomen were most useful . No preponderant coexistent lesion other than gallstones was noted . Levels of carcinoembryonic antigen in gallbladder bile in cases of AMG (2.5 +/- 1.5 ng/ml, mean +/- SD) were significantly lower than in gallbladder carcinoma (p less than 0.01) . All the patients were easily treated with cholecystectomy, and 24 patients who have been followed up after surgery are doing well.(ABSTRACT TRUNCATED AT 250 WORDS) Parazitologiia, 1992 Mar-Apr, 26(2), 105 - 14 {The effect of the vole subspecies of the plague microbe on the viability of Callopsylla caspia fleas}; Vashchenok VS et al.; When infected with cultures of plague microbe reared at 28 C the experimental fleas showed no reliable differences in the mortality as compared to control ones . By contrast, the infection with agent cultures reared at 37 C as well as the infection on agonizing white mice caused a statistically reliable increase in the mortality in insects of both sexes . In the groups of fleas infected on white mice the most intensive mortality was observed during the first 24 hours after the infectious feeding that, apparently, is connected with the effect of toxins present in the blood of agonizing animals. Biotechnology (N Y), 1992 Mar, 10(3), 301 - 4 DnaK-mediated alterations in human growth hormone protein inclusion bodies; Blum P et al.; Protein overproduction in microbes frequently results in protein misfolding and aggregation though the molecular basis for this process is unclear . The HSP70 chaperonin, DnaK, was identified as an important factor controlling heterologous protein aggregation in Escherichia coli . Co-overproduction of DnaK significantly reduced human growth hormone (HGH) protein inclusion body formation and the extent of HGH aggregation. Biotechnology (N Y), 1992 Mar, 10(3), 286 - 91 Transformation of microbes, plants and animals by particle bombardment; Klein TM et al.; Over the past several years, particle bombardment has evolved into a useful tool for molecular biologists, allowing direct gene transfer to a broad range of cell and tissue types . Some of the important applications of the process include the production of transgenic crop species including maize and soybean and the introduction of DNA into plastids and mitochondria . Recent results have extended the range of gene transfer by particle bombardment to animal and bacterial cells . One noteworthy newer application is the direct insertion of genes into the organs of living animals . Here we discuss these advances and the instrument developments that contributed to them. Br J Nutr, 1992 Mar, 67(2), 245 - 55 The effect of sucrose supplements on particle-associated carboxymethylcellulase (EC 3.2.1.4) and xylanase (EC 3.2.1.8) activities in cattle given grass-silage-based diet; Huhtanen P et al.; Carboxymethylcellulase (EC 3.2.1.4; CMCase) and xylanase (EC 3.2.1.8) activities were assayed in rumen fluid and from microbes closely associated either with rumen particulate material or with feed particles incubated in nylon bags in the rumen of cattle . The cattle were fitted with a permanent rumen cannula and a simple 'T'-piece duodenal cannula and were given four diets in a 4 x 4 Latin Square experiment . The basal diet (diet C) consisted of grass silage, barley and rapeseed meal (700, 240 and 60 g/kg total dry matter (DM)) given at the rate of 5.3 kg/d or supplemented with 1.0 kg sucrose/d given twice daily (diet S), twice daily with 0.25 kg sodium bicarbonate/d (diet B) or as a continuous intrarumen infusion (diet I) . Giving sucrose supplements decreased CMCase and xylanase activities extracted from microbes associated with rumen particulate material or feed particles incubated in nylon bags as compared with diet C . Supplementation of the sucrose diet with sodium bicarbonate resulted in higher CMCase and xylanase activities than other sucrose diets (S and I) . Particle-associated CMCase and xylanase activities were found to be very sensitive in detecting differences in the rumen environment and were related to changes in cell wall digestion . The activities were highly correlated with disappearance of DM and neutral-detergent fibre from nylon bags incubated in the rumen, rumen and total digestion of cell-wall carbohydrates and rumen pool size of cell-wall carbohydrates . It was concluded that the attachment of fibrinolytic enzymes is involved in the depression of fibre digestion . Particle-associated CMCase and xylanase activities were much higher when measured from rumen particulate material than from feed particles incubated in nylon bags. J Bacteriol, 1992 Feb, 174(3), 1068 - 71 Extracellular polysaccharide is required for wild-type virulence of Pseudomonas solanacearum; Kao CC et al.; Several Pseudomonas solanacearum strains which produced no detectable extracellular polysaccharide (EPS) in planta had been reported to remain highly virulent when tested at high inoculum concentrations (P . Xu, M . Iwata, S . Leong, and L . Sequeira, J . Bacteriol . 172:3946-3951, 1990; P . Xu, S . Leong, and L . Sequeira, J . Bacteriol . 170:617-622, 1988) . Two of these mutants, KD700 and KD710, have now been molecularly and genetically mapped to the EPSI gene cluster described by Denny and Baek (Mol . Plant-Microbe Interact . 4:198-206, 1991) . When a range of inoculum concentrations was used, these two mutants and all other EPS-defective mutants tested were found to be reduced in virulence to eggplants and tobacco relative to the wild-type strain . Thus, EPS consistently is required for the wild-type level of virulence in P . solanacearum. Curr Opin Immunol, 1992 Feb, 4(1), 3 - 7 Antibiotic peptides as mediators of innate immunity; Zasloff M; Antibiotic peptides are widely distributed in nature . Almost all function as membrane-active agents, disrupting target-cell permeability . Several exhibit a striking selectivity for single-celled microbes over metazoan cells, and as such are amongst the simplest components of the animal's defensive system, which distinguishes environmental microbes from 'self'. Rheum Dis Clin North Am, 1992 Feb, 18(1), 37 - 48 Do bacterial antigens cause reactive arthritis? Granfors K. There are at least three common features among the microbes causing reactive arthritis: (1) they primarily cause infections on mucosal areas, (2) they are intracellularly living microorganisms, and (3) they have lipopolysaccharide as an essential structure of their outer membrane . It is obvious that during or after the acute infection microbial antigens are not properly eliminated, and they persist for long times in HLA-B27-positive persons developing ReA . Bacterial lipopolysaccharide has also been shown to enter the joints and is an important factor in the pathogenesis of ReA. Am J Clin Pathol, 1992 Feb, 97(2), 202 - 8 A flow cytometric method to detect anti-pyruvate dehydrogenase antibody in primary biliary cirrhosis; Elkhalifa MY et al.; Primary biliary cirrhosis (PBC) is an autoimmune disease characterized by the presence of anti-mitochondrial antibodies specifically directed against the M2 group of mitochondrial antigens . Recently, the E-1, the E-2, and protein X components of pyruvate dehydrogenase enzyme complex have been identified as the major antigens within the M2 group of autoantigens . An immunoassay using pyruvate dehydrogenase enzyme complex as a specific antigen for the diagnosis of PBC was developed . Pyruvate dehydrogenase enzyme complex was attached to polystyrene microbeads, incubated with sera from PBC patients (n = 18), normal controls (n = 50), or patients with other autoimmune diseases (n = 26), followed by incubation with a second fluorescein isothiocyanate conjugated goat anti-human immunoglobulin and then analyzed by flow cytometry . High numbers of fluorescence channels (mean, 1,693 +/- 846) were obtained for all PBC sera except for two patients . Compared to the conventional anti-mitochondrial antibody assay, the assay had a sensitivity rate of 94% and a specificity rate of 100% . The reactive antibodies are predominantly of the immunoglobulin G3 subclass . Their levels could be correlated with the histopathologic stages of PBC . These results were corroborated by immunoblotting . Sera from patients with later stages of PBC strongly reacted with pyruvate dehydrogenase enzyme complex components, E1 alpha, and protein X. Arch Surg, 1992 Feb, 127(2), 218 - 21 The Surgical Infection Society's policy on human immunodeficiency virus and hepatitis B and C infection . The Ad Hoc Committee on Acquired Immunodeficiency Syndrome and Hepatitis; Davis JM et al.; The Ad Hoc Committee on Acquired Immunodeficiency Syndrome and Hepatitis of The Surgical Infection Society has outlined its policy regarding three deadly blood-borne viral infections . The risk of transmission of these microbes, the role of preoperative testing, the problem of the human immunodeficiency virus-infected surgeon, and conduct in the operating room are discussed. Khirurgiia (Mosk), 1992 Feb, (2), 94 - 101 {The use of the carbon dioxide laser in acute surgical infection of the soft tissues}; Kuleshov SE et al.; Clinico-laboratory studies were conducted in 46 patients with wound infection for the treatment of which carbon-dioxide laser was used, and in 78 patients who were operated on with a metal scalpel (control) . It is shown that "excision" and "evaporation" of tissues took 15-45 minutes . Repeated interventions were performed in 15% of cases due to bleeding from vessels measuring 1.0-1.5 mm in diameter . After surgical debridement with a scalpel the number of microbes reduced from 10(7)-10(9) to 10(3)-10(4) per g . After exposure to the laser beam this value was 10(1) but on day 3-4 it was 10(5)-10(6) . The extent of the zone of coagulation necrosis in the zone exposed to the laser beams measured up to 500 microns, and began diminishing only on day 7-9 of postoperation . Suppuration of the wounds in 56% of patients of the main group treated by operation was in conformity with the results of bacteriological and morphological studies . In the control group this index was 15.4% . It is concluded that the use of carbon-dioxide laser as a "light scalpel" with subsequent primary closure of the wound is inexpedient . Its use in preparing a wound for closure with secondary sutures has no advantages over radical surgical debridement. J Clin Microbiol, 1992 Feb, 30(2), 377 - 80 Clinical and laboratory analyses of cytospin-prepared Gram stains for recovery and diagnosis of bacteria from sterile body fluids; Chapin-Robertson K et al.; The smear of a clinical specimen provides essential laboratory information that is used to make therapeutic decisions . For this study, smears were made by centrifugation in a Beckman Microfuge 11 (Beckman Instruments, Palo Alto, Calif.) and in parallel by using a Cytospin 2 apparatus (Shandon Inc., Pittsburgh, Pa.) . Of 350 consecutive body fluid specimens examined, 50 (14.0%) grew bacteria . Both methods were culture and smear positive for 24 (6.9%) specimens; 18 (5.1%) specimens were cytocentrifuge smear positive, culture positive, and high-speed centrifugation (HSC) negative; 3 (0.8%) were culture negative and positive by both smear methods; and 1 (0.2%) was HSC smear positive, culture positive, and cytocentrifuge negative . Seven (2.0%) specimens were culture positive and negative by both smear methods . Clinically, cytocentrifuge preparations showed greater sensitivity for culture-positive specimens and a closer correlation with the CFU per milliliter than HSC did, resulting in a greater ability to treat patients with specific therapies . In addition, analysts needed to examine only a 6-mm-diameter area on the slide, cells and microbes were somewhat larger and more regular in appearance, and smears stained more uniformly . Because of the increased clinical and laboratory utility of the cytocentrifuge, its use is recommended in clinical microbiology laboratories for all sterile body fluid specimens. Biotechnology (N Y), 1992 Feb, 10(2), 157 - 62 Rational immunotherapy with interleukin 2; Kaplan G et al.; Interleukin 2 (IL-2), a T lymphocyte product released upon antigen stimulation, has been used for cancer therapy in high doses for more than five years . More recently, its potential as a stimulant of cell-mediated immunity in infectious diseases, particularly those caused by intracellular microbes, has become appreciated . Drawing on the extensive information available as to the structure, cellular and molecular effects of IL-2, this review focuses on its use in patients with lepromatous leprosy and AIDS in low, physiologic doses . The data indicate that IL-2 is effective in stimulating cell-mediated immunity without systemic toxicity. Crit Care Clin, 1992 Jan, 8(1), 163 - 89 Inflammatory host responses in sepsis; Zimmerman JJ et al.; Although microbes and their associated toxins initiate sepsis, it is the subsequent host inflammatory response that defines most of what we characterize as clinical sepsis . This article considers the various cellular as well as humoral mediators involved in this response in addition to the complex networking that may result in both augmentation and modulation of the inflammatory response in sepsis. Invest Ophthalmol Vis Sci, 1992 Jan, 33(1), 121 - 5 Analysis of glycoprotein deposits on disposable soft contact lenses; Tripathi PC et al.; By using gel electrophoresis, as well as Western blotting with specific antibodies or with the lectin concanavalin A, we characterized the types and amounts of proteins that are deposited on 58% ionic and 38% nonionic water-content disposable soft contact lenses (DSCLs) worn for 1 to 21 days by asymptomatic subjects with mild to moderate myopic refractive errors . The total amounts of protein eluted from the lenses ranged from 0.1 to 80 micrograms/lens . The amount of protein deposited on 58% water-content lenses was greater than that on 38% water-content DSCLs . We did not find a strict correlation between the amount of protein deposited and the duration of wear for either type of lens . The major polypeptide fractions detected had apparent molecular weights of 14, 17, 21, 30, and 60 kD . The fractions at 14 kD-bound antibodies specific for human lysozyme, and those at 17 kD corresponded to prealbumin . The 60 kD fraction included IgG heavy chains . The identity of the fractions at 21 kD and 30 kD is unknown . Because oligosaccharide side chains on the proteins attract microbes and facilitate their adherence, knowledge about the types of carbohydrate moieties in lens deposits can provide a rational approach to inhibiting or reversing microbial infection. Tsitologiia, 1992, 34(2), 62 - 7 {The effect of the UV microirradiation of the centrosome on cell behavior . III . The ultrastructure of the centrosome after irradiation}; Uzbekov RE et al.; One of the spindle poles of mitotic PK cells was irradiated with UV microbeam in metaphase or in anaphase . Electron microscopy showed that immediately after irradiation the microtubules around the centrosome were maintained, and that the ultrastructure of both irradiated and nonirradiated poles was similar . After microirradiation of the centrosome in metaphase, the mitotic halo around this centrosome was retained, but in due time the number of microtubules was getting less compared to that around the nonirradiated centrosome . When daughter cells with irradiated centrosomes are passing into the interphase, their centrioles are not separated from each other, no primary cilia are formed, and no replication of centrioles occurs . In the interphase cells with irradiated centrosomes, satellites are formed on the active centriole, but centrosome-attached microtubules are practically absent. Lasers Surg Med, 1992, 12(1), 51 - 62 Microbeam laser-injured neurons increase in vitro astrocytic gap junctional communication as measured by fluorescence recovery after laser photobleaching; Anders JJ et al.; An important aspect of the neuronal-astrocyte relationship is the interaction of reactive astrocytes with injured and/or dying neurons . Few studies have focused on the signaling of astrocytes by injured neurons or on the possibility that neurons can alter astrocytic gap junctional communication . The purpose of this study was to determine whether the presence of injured neurons could alter astrocytic gap junctional coupling by establishing an in vitro method of microbeam laser neuronal injury and coculturing these neurons with astrocytes . Neurons from two rat neuronal clonal cell lines were injured using a 20-W argon laser operating on the ultraviolet (UV) multiline (351-361 nm) directed through a X40 objective of an inverted microscope . After laser injury, the glass slide with the injured neurons was sandwiched with a slide on which primary rat astrocytes were grown . Although the neurons and astrocytes were bathed in the same medium, they were not in direct contact during the coculture period (24, 48, or 72 hr) . Astrocytic gap junctional dye coupling was examined using the fluorescence recovery after laser photobleaching (gap-FRAP) analysis technique . Astrocytes cocultured with the injured neurons for 24 to 48 hr did not show a significant difference in fluorescence recovery when compared to control values . After 72 hr of coculture, there was a significant increase in the gap junctional dye coupling . These results indicate that injured neurons influence in vitro astrocytic gap junctional conductance after 72 hr of coculture as measured by dye coupling. Annu Rev Immunol, 1992, 10, 809 - 34 Complement deficiencies; Colten HR et al.; The complement system consists of about two dozen plasma and cell membrane proteins which function as cofactors in defense against pathogenic microbes and in the generation of many immunopathogenic disorders . The complexity of this system and its role in other biological functions has been appreciated within the last two decades . Recognition of genetic deficiencies of the complement proteins and their phenotypic expression has provided additional insights into the physiological role of the complement system . Complementary DNA (cDNA) clones for most of the complement components are now available, and the gene structures for many have been elucidated . Application of molecular biological methods to studies of the complement system and its deficiencies has permitted both the determination of primary structure and chromosomal localization of complement genes (Figure 1) and the capacity to elucidate the molecular basis of complement deficiency disorders. Pharm Acta Helv, 1992, 67(1), 15 - 9 Bioconversion of tamoxifen by microbes; el-Sharkawy SH; Screening studies of tamoxifen (TAM) have shown that only one culture, Streptomyces rimosus ATCC 2234 was able to metabolize tamoxifen to 4-hydroxy tamoxifen (OHT) which was obtained in a 10% yield and chemically identified by 2 D nmr spectroscopy . A number of microorganisms were capable of biotransforming TAM to tamoxifen-N-oxide (TNO), and desmethyltamoxifen (DMT) . The chemical identity of the isolated metabolites was achieved using gas chromatography/mass spectrometry and different nmr spectroscopic techniques. Appl Environ Microbiol, 1992 Jan, 58(1), 99 - 105 Comparative growth rates of various rumen bacteria in clarified rumen fluid from cows and sheep fed different diets; van Glyswyk NO et al.; Pure cultures of strains of different species of rumen bacteria were grown in filter-sterilized rumen fluid supplemented with glucose, bicarbonate, and reducing agent (cysteine and sulfide) . Growth rates were determined in a series of experiments . Strains of species most abundant in the rumen grew more rapidly than strains of less abundant bacteria . Ammonia, amino acids, and peptides increased growth rates to some extent, but the greatest stimulatory effect for less abundant bacteria was provided by other factors, present in yeast extract . Factors released from lysates of mixed rumen microbes stimulated growth, but their rate of release was slow . It was concluded that, besides energy and nitrogen sources, growth factors of an as-yet-undetermined nature probably play an important role in determining the predominance of different bacterial species in the rumen. J Inherit Metab Dis, 1992, 15(4), 683 - 6 Biochemistry and molecular biology of chronic granulomatous disease; Segal AW; The NADPH oxidase of phagocytic cells is an electron transport chain in the wall of the endocytic vacuole that is activated upon engulfment of the microbe, and is important for its efficient killing and digestion . The oxidase consists of a heterodimeric cytochrome b in the membrane, which is the site of the haem and FAD groups, and two cystolic factors p47-phox and p67-phox that appear to activate the system . Absence of this oxidase as a result of defects in each of these specialized proteins causes the syndrome of chronic granulomatous disease (CGD), that is characterized by a profound predisposition to pyogenic infection. Microbios, 1992, 70(282), 13 - 22 Aluminium, chromium and manganese detoxification mechanisms in Pseudomonas syringae: an X-ray fluorescence study; al-Aoukaty A et al.; Pseudomonas syringae cultured in a defined citrate medium supplemented with 1 mM aluminium, chromium and manganese, respectively, appeared to elicit disparate biochemical responses . At the stationary phase of growth aluminium was predominantly present as an insoluble residue . Although virtually none of this metallic element was detected in the supernatant, the bacterial cells appeared to contain some aluminium . Following the initial uptake of chromium the microbe secreted the metal in the supernatant . Only a small fraction of the chromium was localised in the bacterial cells; 91% manganese was biotransformed into an insoluble pellet . No citrate was detected in the exocellular fluid at cessation of cellular growth. Pharmacol Ther, 1992, 55(3), 201 - 77 AIDS acquired by drug consumption and other noncontagious risk factors; Duesberg PH; The hypothesis that human immunodeficiency virus (HIV) is a new, sexually transmitted virus that causes AIDS has been entirely unproductive in terms of public health benefits . Moreover, it fails to predict the epidemiology of AIDS, the annual AIDS risk and the very heterogeneous AIDS diseases of infected persons . The correct hypothesis must explain why: (1) AIDS includes 25 previously known diseases and two clinically and epidemiologically very different epidemics, one in America and Europe, the other in Africa; (2) almost all American (90%) and European (86%) AIDS patients are males over the age of 20, while African AIDS affects both sexes equally; (3) the annual AIDS risks of infected babies, intravenous drug users, homosexuals who use aphrodisiacs, hemophiliacs and Africans vary over 100-fold; (4) many AIDS patients have diseases that do not depend on immunodeficiency, such as Kaposi's sarcoma, lymphoma, dementia and wasting; (5) the AIDS diseases of Americans (97%) and Europeans (87%) are predetermined by prior health risks, including long-term consumption of illicit recreational drugs, the antiviral drug AZT and congenital deficiencies like hemophilia, and those of Africans are Africa-specific . Both negative and positive evidence shows that AIDS is not infectious: (1) the virus hypothesis fails all conventional criteria of causation; (2) over 100-fold different AIDS risks in different risk groups show that HIV is not sufficient for AIDS; (3) AIDS is only 'acquired,' if at all, years after HIV is neutralized by antibodies; (4) AIDS is new but HIV is a long-established, perinatally transmitted retrovirus; (5) alternative explanations disprove all assumptions and anecdotal cases cited in support of the virus hypothesis; (6) all AIDS-defining diseases occur in matched risk groups, at the same rate, in the absence of HIV; (7) there is no common, active microbe in all AIDS patients; (8) AIDS manifests in unpredictable and unrelated diseases; and (9) it does not spread randomly between the sexes in America and Europe . Based on numerous data documenting that drugs are necessary for HIV-positives and sufficient for HIV-negatives to develop AIDS diseases, it is proposed that all American/European AIDS diseases, that exceed their normal background, result from recreational and anti-HIV drugs . African AIDS is proposed to result from protein malnutrition, poor sanitation and subsequent parasitic infections . This hypothesis resolves all paradoxes of the virus-AIDS hypothesis . It is epidemiologically and experimentally testable and provides a rational basis for AIDS control. Vox Sang, 1992, 63(4), 272 - 5 An enzyme-linked antiglobulin test to quantify nanogram quantities of IgG on polystyrene microspheres; Greenwalt TJ et al.; A method is described in which polystyrene latex beads are used for constructing standard curves to estimate the number of protein molecules on cell surfaces by an enzyme-liked immunoassay test (ELAT) . A series of immune globulin (IgG) dilutions in pH 9.8 carbonate buffer were coated on 3-microns microbeads by incubation overnight at 4 degrees C and subjected to ELAT . The r value of the curve derived from four assays was 0.9991 . This standard curve applied to previously recorded ELAT data resulted in estimating that normal RBC have 63 +/- 19 (SD) IgG molecules and that the lower level of sensitivity of the antiglobulin test is 155 IgG molecules per RBC . The method should be useful for more precise standardization of procedures for measuring proteins on cell surfaces. Biomaterials, 1992, 13(13), 949 - 52 Chitosan beads and granules for oral sustained delivery of nifedipine: in vitro studies; Chandy T et al.; Nifedipine was embedded in a chitosan matrix to develop a prolonged-release form . The in vitro release profiles of nifedipine from chitosan beads and microgranules were monitored by UV spectrophotometer . The studies were performed in a rotating shaker (100 rev min-1) in 0.1 M HCl buffer (pH 2.0) or 0.1 M phosphate buffer (pH 7.4) . Comparison was made between drug-loaded microbeads and microgranules . The amount and percentage of drug release were much higher in HCl than in phosphate buffer, probably due to the salt formation of the matrix (chitosan hydrochloride) at acid pH . The release rate of nifedipine from chitosan matrix was slower for beads than granules . These findings suggest the possibility of modifying the formulations to obtain the desired controlled release of the drug in an oral sustained-delivery system. Annu Rev Microbiol, 1992, 46, 219 - 52 Biodiversity as a source of innovation in biotechnology; Bull AT et al.; The object of this article is to draw attention to the significance of microbial diversity as a major resource for biotechnological products and processes . The topic is approached from two complementary standpoints . First, an attempt is made to assess the extent of biodiversity, particularly microbial diversity . In this context, the application of the modern techniques of molecular biology is enabling the detection of hitherto completely unknown groups of microbes and, also, is revealing the extent of genetic diversity within microbial taxa . The case is made for the establishment of sound microbial taxonomies both on the basis of satisfying fundamental scientific needs, and for designing effective isolation strategies . The impact of an ecological approach to search and discovery of novel organisms and properties also is emphasized and illustrated . Second, the question of screening a collection of appropriate microorganisms for the desired attributes is considered . The focus here is placed on modern intelligent or targeted screening, and on the power of molecular biology to extend the range of screening options . Discussions of microbial ecology or diversity only rarely touch upon questions of gene pool conservation . The point made here is that loss of biodiversity should be as ominous for microbiologists and biotechnologists as it is to conservationists . The article concludes with thoughts on some means of conserving microbial diversity. Przegl Lek, 1992, 49(5), 141 - 4 {Chlamydia trachomatis infection of the urogenital system in our clinical experience}; Magott M et al.; The increased number of the genitourinary system infection caused by Chlamydia trachomatis (Ch . Tr.), increased number of patients with dysuria or sterile leukocyturia gave stimulus to studies of 615 patients from Department of Nephrology and District Outpatient Nephrological Care Unit with regard to infections with that microbes . Material for investigations derived from urethra . Diagnostic examinations were performed using the Mc Coy cell culture and the immunofluorescence method . The infection was noted in 176 patients (119 women and 57 men) that is in 28.6% of cases studied . The mean age of patients was 42.7 +/- 12 years . Clinical symptoms such as dysuria or frequency were typical for that kind of infection . The most frequent abnormality was leukocyturia or leukocyturia accompanied by erythrocyturia noted in 66% of patients . Isolated erythrocyturia was observed in 24.4% of cases . It has been stated that anamnesis or routine laboratory examinations were not able to the identification of infection . In face of poorly characteristics of clinical picture of infection the infection with Ch.Tr . could be the cause of unsuccessful therapy in patients with signs of genitourinary tract infections. Dysphagia, 1992, 7(4), 226 - 33 Velar activity and timing of eustachian tube function in swallowing; Hamlet SL et al.; Velar motion for dry and liquid swallows was investigated . as well as velar activity in speech, based on X-ray microbeam pellet tracking data . Electromyographic recordings for tensor and levator veli palatini were obtained simultaneously . Velar pellet trajectories for swallowing were more complex than for speech, since there was a high-velocity anterior component in swallowing . For some swallows this anterior component was integrated with velar elevation (especially in liquid swallows), but in other cases initial velar elevation occurred considerably earlier (chiefly in dry swallows) . The burst of tensor and levator veli palatini activity characteristic of swallowing was associated with the anterior component of velar pellet motion, but not consistently with velar elevation per se . The conventional view on timing of tensor veli palatini contraction in a swallow, which governs Eustachian tube opening, is that this is associated with velar closure . The X-ray microbeam data suggest rather that Eustachian tube ventilation is more closely associated in time to the onset of pharyngeal peristalsis, which may or may not coincide with initial velar elevation. Basic Life Sci, 1992, 59, 915 - 34 Condensed tannins in some forage legumes: their role in the prevention of ruminant pasture bloat; Lees GL; For the past 20 years, the focus in our laboratory has been on finding the causes of ruminant pasture bloat and eventually breeding a bloat-safe alfalfa (Medicago sativa L.); i.e., with bloat potential reduced to the economic threshold . In the mid-seventies, the mechanisms of bloat were explored and found to be more physical than chemical . Characteristic of all bloating legumes after ingestion was a very rapid initial rate of ingestion by rumen microbes . Through the study of bloating and non-bloating legumes, factors were elucidated in the plant that would slow this process . One of these factors was the presence of condensed tannins in the herbage . Some of the non-bloating legumes contained these secondary metabolites, but no condensed tannins were found in any of the bloating legumes . Therefore, species containing an appreciable amount of condensed tannins in their leaves and stems are considered to be non-bloating . Conventional breeding methods have not been successful in producing an alfalfa with condensed tannins in its herbage . New approaches using tissue culture techniques are being attempted, but genetic engineering has the greatest potential for success. Eicosanoids, 1992, 5(1), 5 - 8 Comparison of bioanalytical determinations of Iloprost, a chemically stable PGI2 mimetic, by conventional radioimmunoassay (RIA) and scintillation proximity assay (SPA); Hildebrand M et al.; The scintillation proximity assay is a novel variant of classical radioimmunoassay . It can be performed as a single tube measurement because the separation of bound and unbound tracer fraction is avoided . In principle, microbeads are coated with anti-species antibodies that can couple with the respective antiserum used for RIA . By means of special cores, light emission takes place if labelled, antiserum-bound tracer is coupled to the anti-species antibody on the fluomicrosphere surface . In the present report, the novel assay was compared to a validated RIA for the bioanalysis of the PGI2 mimetic, Iloprost . Extraction recovery of Iloprost was approximately 90% at pH less than or equal to 4 . The detection limit of the novel assay was 2-4 pg/sample, corresponding to 10-20 pg/ml plasma (if 0.2 ml plasma was used) . Coefficients of variations were 9, 7 and 6% (within-day, n = 5) and 30, 11 and 10% (day-to-day, n = 10) at 50, 100 and 200 pg/ml . RIA and SPA levels of Iloprost measured in human plasma samples (n = 428) were similar . The SPA method exhibits both a similar specificity and detection limit to RIA and will be used for further analyses. Mol Carcinog, 1992, 5(4), 301 - 10 Homologous and heterologous gap-junctional intercellular communication in v-raf-, v-myc-, and v-raf/v-myc-transduced rat liver epithelial cell lines; Kalimi GH et al.; We examined gap-junctional intercellular communication (GJIC) in a series of normal and v-raf-, v-myc-, and v-raf/v-myc-transduced rat liver epithelial (RLE) cell lines using the scrape loading-dye transfer and fluorescence-recovery-after-photobleaching (FRAP) assays . Whereas the normal RLE cell line, the control helper virus-transduced cell line, and the v-myc-transduced cell line all showed excellent GJIC, the v-raf-transduced cell lines displayed decreasing levels of GJIC associated with their increasing tumorigenicity . The v-raf/v-myc-transformed cell lines showed the lowest levels of GJIC and were also the most tumorigenic . Heterologous GJIC of these oncogene-transduced cell lines was also compared with that in the normal RLE cells . A modified FRAP assay, using fluorescent-microbead labelling to identify the oncogene-transduced cell from surrounding normal cells, was used to quantify the heterologous GJIC . The v-raf/v-myc-transformed RLE cells had no heterologous communication with the normal RLE cells, whereas v-raf- and v-myc-transduced cell lines maintained heterologous GJIC . Northern analysis showed that connexin 43 was the only gap-junction protein message expressed in these cell lines; connexin 32 and connexin 26 were not expressed . The levels of connexin 43 mRNA expression were relatively unchanged in all cell lines, suggesting that the reduction in GJIC was primarily at the posttranslational level . These findings suggest that reduction of homologous GJIC in v-raf- and v-raf/v-myc-transformed RLE cells is linked to their tumorigenic potential . Furthermore, the loss of heterologous GJIC, which we observed only in the v-raf/v-myc-transformed cells, might release such cells from the growth-regulating effects of surrounding normal cells, possibly contributing to their enhanced tumorigenic potential. Res Immunol, 1992 Jan, 143(1), 71 - 8 Endotoxin-neutralizing capacity of soluble CD14; Schutt C et al.; Luminol-enhanced chemiluminescence was used to determine the effect of soluble CD14 (sCD14) on the endotoxin-inducible generation of reactive oxygen species in human monocytes . It was necessary to mediate lipopolysaccharide (LPS) monocyte-activating capability by serum factors (LPS-binding proteins) . sCD14 reduced LPS-inducible monocyte activation in a dose-dependent manner, even in the case of CD14- monocytes, obtained from a patient with paroxysmal nocturnal haemoglobinuria . These monocytes could be activated by opsonized LPS via other receptors . Using anti-mouse Ig-coated microbeads, it was demonstrated in FACS analysis that sCD14 mediates the binding of a mouse monoclonal anti-CD14 antibody (RoMo 1) to a complex of LPS/FITC (fluoroisothiocyanate) and a LPS-binding protein . The release of sCD14 from cultured monocytes was measured using LPS, TNF alpha (tumour necrosis factor), IL1, 4 and 6 (interleukin-1, -4 and -6) and IFN gamma (interferon-gamma) as stimulators . Addition of LPS and TNF alpha led to a dose-dependent increase in sCD14-levels in the culture supernatant, whereas IL1, IL6 and IFN gamma had no significant effect . IL4 dose-dependently depressed spontaneous sCD14 release . It is possible that elevated sCD14-serum levels in polytraumatized patients indicate a natural protective mechanism against excessive monocyte mediator production . Therefore, sCD14 may be a new therapeutic concept in endotoxic shock prevention. J Appl Physiol, 1992 Jan, 72(1), 1 - 7 Tissue-specific regulation of inflammation; Colten HR; Proteins of the complement system are important effectors and modulators of inflammation . The complement cascade is triggered by microbes, tissue debris, and specific antibodies . Serum complement proteins are derived primarily from liver, but extrahepatic complement synthesis is important in homeostasis and in local host defenses . Tissue-specific regulation of expression of complement genes is governed by mechanisms similar to those that regulate other "acute phase reactants." That is, tissue injury or infection elicit changes in expression of these acute phase proteins, which, although variable in kinetics, magnitude, and direction, are a consequence of an elaborate system of cell-to-cell communication . This communication is mediated via a complex network of cytokines, including the interferons, interleukins, several growth factors, and sex hormones . The cell biological and molecular biological details of these mechanisms are now under active investigation . An understanding in molecular terms of the balance between proinflammatory and counterregulatory forces on complement gene expression should provide new insight into the functions of complement and the design of novel therapies for disorders of inflammation. Biodegradation, 1992, 3(1), 3 - 18 Microbes and microbial enzymes for cyanide degradation; Raybuck SA; Cyanide is an important industrial chemical produced on a grand scale each year . Although extremely toxic to mammalian life, cyanide is a natural product generated by fungi and bacteria, and as a result microbial systems have evolved for the degradation of cyanide to less toxic compounds . The enzymes which utilize cyanide as a substrate can be categorized into the following reaction types: substitution/addition, hydrolysis, oxidation, and reduction . Each of these categories is reviewed with respect to the known biochemistry and feasibility for use in treatment of cyanide containing wastes. APMIS Suppl, 1992, 27, 71 - 83 Microbial interaction with animal cell surface carbohydrates; Karlsson KA et al.; Microbes have selected primarily carbohydrates for attachment to host animal cells . Recent studies have revealed essential characteristics in the recognition of receptor carbohydrates . Of importance is the property of recognizing also sequences placed inside an oligosaccharide chain, which differs from most animal antibodies . This is the basis for series of isoreceptors with the minimum receptor sequence in common but with separate neighbouring groups . There are families of microbial ligands that show different preferences for members within one series of isoreceptors, indicating only slight differences in the complementary binding sites of the proteins . Such differences may explain shifts in the selectivity of separate host tissues for infection . A second characteristic is the low affinity interaction often found where simple receptor-containing saccharides are unable to inhibit attachment . Technical possibilities are rapidly developing for the design of synthetic receptor analogues to be used in the therapy of clinical infections . This is urgently needed in cases where no rational therapy exists today. Monatsschr Kinderheilkd, 1992 Jan, 140(1), 6 - 18 {Encephalitis . Recent clinical, diagnostic and therapeutic aspects}; Hirt HR; The subjects of this paper are mainly the encephalitis of viral origin and in addition 3 types of non-viral encephalitis of practical importance . A review is given on the current diagnostic possibilities including; clinical criteria; examination of the cerebrospinal fluid; neuroradiology and; etiological investigation by means of direct identification of the microbes in the cerebrospinal fluid or by measuring intrathecally produced antibodies . After some general statements concerning treatment the specifically treatable encephalitis caused by Herpes simplex virus, Mycoplasma pneumoniae, Borrelia burgdorferi and Toxoplasma gondii are discussed in more detail. Sci Prog, 1992, 76(300 Pt 2), 139 - 48 Designing microbes for release into the environment; Molin S; After 20 years in which gene technology has become an important part of modern biotechnology we have seen very beneficial applications of the new techniques in the pharmaceutical industry . We are now entering a second phase involving the deliberate release of genetically engineered organisms into the environment . This next step causes concern because of a low level of predictability of their possible effects . While the risk assessment of microbial release is far from easy, the strain designers also face problems concerning optimization of performance of the organisms . The two groups of actors in this new development--the risk assessors and the strain designers--need the same platform of understanding from the field of microbial ecology, and a number of specific areas which may now be approached by modern technology deserve particular attention . An increased understanding of the activities of microbes in the environment will also allow construction of more predictable, and therefore safer, strains . Biological containment and molecular microbial ecology are two sides of the same coin in the context of release of genetically engineered microorganisms. Nat Toxins, 1992, 1(3), 197 - 208 Ecology of plant-herbivore communities: a fungal component? Hammon KE, Faeth SH. We consider how microorganisms may alter conventional theories of the organization of plant-herbivore communities . We focus on endophytic fungi and their role in mediating interactions among herbivores, their host plants, and natural enemies . We propose hypotheses about the role of microbes in plant-herbivore communities and suggest ways to test these hypotheses . An initial approach to the overwhelming complexity of interacting species is to view species as components of functional groups, be they micro- or macroscopic, that potentially affect the ecology and evolution of host plants. Cell Transplant, 1992, 1(5), 349 - 54 Embedded adrenal cells graft reduced local and early nonspecific inflammatory phenomena which follow agarose beads implantation; Cadic C et al.; Microencapsulation of adrenal cells is proposed for reducing the nonspecific inflammatory reaction observed around polymer implants . This hypothesis was tested by comparing both host cellular reaction and the surrounding graft cell populations which appeared either when agarose embedded cells or when empty agarose beads were implanted . Our results showed that the fibrotic material that surrounded the implanted empty agarose microbeads was not as severe and important when adrenal cells were present . Similarly, T lymphocyte population surrounding the graft was considerably reduced together with the percentage of CD4 and CD8 positive cell subpopulations . The activation macrophage marker IaD disappeared . Our results support the hypothesis that embedded adrenal cells may be a suitable solution for reducing early inflammatory events due to microcapsules implantation. Insect Mol Biol, 1992, 1(1), 25 - 36 Phylogeny of cytoplasmic incompatibility micro-organisms in the parasitoid wasp genus Nasonia (Hymenoptera: Pteromalidae) based on 16S ribosomal DNA sequences; Breeuwer JA et al.; Cytoplasmic incompatibility results in embryo mortality in diploids, or all male offspring in haplodiploids, when individuals carrying different cytoplasmic factors are crossed . Cytoplasmic factors have been identified as intracellular micro-organisms . Microbe-induced cytoplasmic incompatibility is found in many insect taxa and may play a role in reproductive isolation between populations . Such micro-organisms cause bidirectional incompatibility between species of the parasitoid wasp genus Nasonia . The phylogenetic relationship of cytoplasmic incompatibility microorganisms (CIM) of different Nasonia species was analysed using their 16S ribosomal DNA (rDNA) sequence . Two 16S rDNA operons were detected in the CIM of each Nasonia species . Sequence analysis indicates that the Nasonia CIM are closely related and belong to the alpha group of the Proteobacteria. Zh Mikrobiol Epidemiol Immunobiol, 1992, (11-12), 5 - 7 {Phosphatase and penicillinase activities as stable traits for the differentiation of the racial classification of Francisella tularensis}; Pavlovich NV et al.; In the causative agent of tularemia new markers correlating with different subspecies of this microbe have been detected . Thus, F . tularensis strains belonging to the American and Central Asian subspecies are characterized by phosphatase activity, which makes it possible to use the phosphatase test for their differentiation from the strains of the holarctic variety . F . tularensis subsp . mediasiatica are incapable of producing beta-lactamase which differentiates them from the representatives of the varieties holarctica and tularensis . These newly discovered signs are stable and do not depend on the virulence of the cultures under study and on the conditions of the cultivation of F . tularensis. Ciba Found Symp, 1992, 171, 184 - 94; discussion 195-8 Roles of secondary metabolites from microbes; Vining LC; The common feature of the seemingly diverse array of biological activities exhibited by microbial secondary metabolites is their survival value for the producing organism . The propensity to form these compounds is unevenly distributed in microbial taxa and seems more closely associated with existence in a competitive environment than with phylogeny . The characteristic multibranched elaboration of secondary biosynthetic pathways and the marked species specificity of the end products are consistent with their evolution by an 'inventive' mechanism . The species specificity suggests that distinctive terminal reactions may be of recent origin . However, comparisons of the nucleotide sequence of genes involved in the biosynthesis of phenazine and polyketide metabolites with related genes of primary pathways indicate that the secondary pathways have not evolved exclusively within the organisms in which they are now found . Sequence similarities with related primary pathway genes in phylogenetically distant organisms suggest that gene transfer has played an important part in the evolution of secondary metabolism . The diversity of products may reflect the many roles for which secondary metabolites have been selected after the genes for their biosynthesis have transferred to organisms with different physiologies and different environment challenges. Arch Immunol Ther Exp (Warsz), 1992, 40(3-4), 183 - 90 Macrophage activating factors produced in the course of murine tularemia: effect on multiplication of microbes; Kovarova H et al.; Primary F . tularensis infection in mice induces the production of macrophage activating factors (MAFs) by spleen cells . The stimulation of macrophage cytolytic activity (MAF-c) and hydrogen peroxide production (MAF-H2O2) dominates between days 7 and 10 in the course of tularemia . Three various pools of active fractions (10-11, 14-15, 25-28) were fractionated by two-step chromatography . Typical for 10-11 and 14-15 is MAF-c activity whereas in 25-28 prevails MAF-H2O2 . Initial concentrated supernatant (day 7 of infection) and individual fractions have been used to raise antibodies KI (anti 10-11) and KII (anti 14-15) . Neutralization reactions with specific antibodies indicate the presence of tumor necrosis factor alpha (TNF alpha) in 14-15 (44% inhibitable), interferon gamma (IFN gamma) and interleukin 2 (IL 2) in 25-28 (65% and 30% neutralization, respectively) . Utilizing KI and KII, 99% and 90% inhibition of cytolytic activity is reached in 10-11 and 14-15, respectively, in spite of non-specific cross reaction . Western blot analysis of proteins in supernatant on day 7 detects, besides TNF alpha, further protein bands (13, 15.5, 52 and 72 kDa) that seem to be associated with macrophage activation . Significant protective effect against in vivo multiplication of tularemic microbes indicates a certain role of TNF alpha, however, cooperation of other molecules is worth to be taken into consideration. Arch Tierernahr, 1992, 42(3-4), 197 - 211 {Comparison of methods for determining the prececal nutrient digestibility and amino acid absorption of swine with ileorectal anastomoses and ileocecal bridge cannulas}; Hennig U et al.; The apparent precaecal nutrient digestibility and amino acid (a.a.) absorption of 10 diets were estimated in pigs with end-to-side ileorectal anastomoses (IRA) as well as with ileocaecal re-entrant cannulae (IZB) . The digestibilities of organic matter and crude carbohydrates were significantly higher in IRA pigs than in IZB pigs in two resp . three diets only . There were no practical important differences for the digestibilities of crude protein and crude fat in all diets . Out of the 180 a.a . absorption comparisons only 20 (i.e . 11%) showed differences larger than 5%-units . The absorption rates of several a.a . were lower in IRA than in IZB pigs and of some other a.a . on the contrary . The methionine absorption of five legume diets and one barley + lysine diet were up to 15% resp . 4%-units lower in IRA pigs . Possibly this was due to the activity of microbes in the rectum in connection with a small intake level of protein bound methionine . In order to minimize such a falsification it is proposed to use the end-to-end anastomosis . The deviations in the apparent a.a . absorption between IRA and IZB pigs can be explained partly by differences in the quantity of precaecally excreted endogenous a.a . per 100 g DM intake. Klin Wochenschr, 1991 Dec 15, 69(21-23), 975 - 80 The involvement of oxygen radicals in microbicidal mechanisms of leukocytes and macrophages; Roos D; Phagocytic leukocytes generate large amounts of reactive oxygen compounds during and after phagocytosis of micro-organisms . These compounds are essential for the killing of a wide variety of microbes . The enzyme responsible for this process is NADPH:O2 oxidoreductase (NADPH oxidase), which utilizes the reduction equivalents of NADPH to reduce atmospheric oxygen to superoxide (O2-.) . Subsequently, superoxide is converted by the leukocytes to other reactive compounds, such as hydrogen peroxide (H2O2), hypochlorous acid (HOCl) and N-chloramines (RNCl) . Each of these compounds has potent microbicidal properties . Under resting, non-phagocytizing conditions, phagocytes do not produce reactive oxygen compounds . However, within 15-30 sec after binding of micro-organisms to cell surface receptors, superoxide generation starts . This phenomenon is called the respiratory burst . This phenomenon is called the respiratory burst . The activation of the NADPH oxidase is caused by the assembly of components of this enzyme into an active complex . Under resting conditions, at least three components reside in the cytoplasm and at least two are located in the plasma membrane . Activation of the NADPH oxidase results in translocation of cytosolic components to the plasma membrane and formation of an active enzymatic complex in the plasma membrane. Sci Rep Res Inst Tohoku Univ {Med}, 1991 Dec, 38(2-4), 83 - 9 beta-Lactamase activity in sputum and indirect pathogenicity; Watanabe A et al.; beta-Lactamase activity of sputum of 39 patients with respiratory infections and that of bacterial isolates from their sputum were determined by nitrocefin method . Forty-six microbial strains (16 different species) were recovered from 39 sputum samples . The beta-lactamase activity was negative only in nine of the 46 strains . beta-lactamase-positive strains were recovered from 35 of 39 sputum samples . Of these 35 sputum samples, the beta-lactamase activity was positive in 23 and negative in 12 . beta-lactamase-negative strains were recovered from 4 of 39 sputum samples, and the beta-lactamase activity was negative in all of these four sputum samples . Both beta-lactamase producing and non-producing strains were recovered in 5 of 6 sputum samples from which two to three microbes were isolated . The ratio of beta-lactamase-positive sputum sample was higher in patients who had received antibiotics than in those who had received no antibiotics . This difference was statistically significant (P < 0.025) . It is possible that non-causative beta-lactamase-positive organisms enhance the pathogenicity of causative organisms that are beta-lactamase-negative. J Cell Sci, 1991 Dec, 100 ( Pt 4), 771 - 80 Assembly of actin-containing cortex occurs at distal regions of growing neurites in PC12 cells; Sanders MC et al.; Although actin filaments are known to be localized in the cortex of axons and in the growth cones of nerve cells, it is unclear how actin-containing structures are assembled during nerve growth . We have studied the formation of actin structures in growing neurites by microinjecting fluorescent phalloidin or actin into PC12 neuron-like cells to label endogenous actin filaments . Upon stimulation of neurite growth in cells microinjected with fluorescent phalloidin, little or no fluorescence was detected in nascent growth cones and adjacent neurites despite the presence of actin filaments in these regions, suggesting that actin filaments were primarily formed by de novo assembly rather than the transport and reorganization of pre-existing, phalloidin-labeled actin filaments . Time-lapse observations of the distribution of phalloidin-labeled actin filaments during neurite elongation confirmed that fluorescence associated with pre-existing neurite cortex spread out more slowly than the elongation of neurites . Furthermore, when a dark spot was photobleached with a laser microbeam along neurites of cells microinjected with either fluorescent phalloidin or actin, the spot showed no appreciable translocation during active neurite elongation . Taken together, these results suggest that de novo assembly of actin filaments plays a crucial role in the formation of growth cones and adjacent cortex in the distal region of neurites, but does not appear to require the anterograde or retrograde transport of cortical filaments, or the passive stretching of the proximal segment of the neurite cortex. Biochem Int, 1991 Dec, 25(6), 1043 - 9 Identification of emodinanthrone oxygenase in fungus Aspergillus terreus; Fujii I et al.; Emodinanthrone oxygenase, which catalyzes the oxidation of emodinanthrone to emodin, has been identified in fungus, Aspergillus terreus . The fixation of an oxygen atom at the C-10 position of emodinanthrone from molecular oxygen catalyzed by the enzyme was proved by the 18O2 incubation experiment and analyses of the product emodin by mass spectrometry and nuclear magnetic resonance . The fact that the reaction did not require any foreign electron donor suggested the involvement of internal monooxygenase . Emodinanthrone oxygenase activities were fo