Mutat Res, 1989 Jan-Feb, 225(1-2), 33 - 9 Spontaneous revertants in modified S . typhimurium mutagenicity tests employing elevated numbers of the tester strain; Georghiou PE et al.; It has been proposed that increases in the number of bacteria applied to each plate can enhance the sensitivity of the Ames S . typhimurium mutagenicity assay . These procedures have the potential to elevate the number of spontaneous revertants (SR) by increasing the contribution of pre-existing revertants (PER) present before application of the bacteria to the limited histidine test plates . We have investigated the contribution of PER when 10(9) bacteria are applied to the plates and found that the number of PER is dependent on the handling and storage of the cultures used to inoculate the overnight broth . The average number of PER/10(9) viable bacteria after overnight growth in broths inoculated from a frozen permanent, lyophilized permanent, master plate, and an isolated colony, of TA98 were 267, 188, 57 and 13 respectively . The resultant elevation of the number of SR for a strain may result in a failure to identify a mutagenic response . It is recommended that the number of PER be monitored in any modification of the Ames test that makes use of elevated numbers of bacteria. J Invest Dermatol, 1989 Jan, 92(1), 13 - 7 Immunohistochemical demonstration of immunoglobulin A in human sebaceous and sweat glands; Metze D et al.; Immunoglobulin A (IgA) mediated humoral defense mechanisms have been detected on all mucous membrane surfaces . There are only a few papers about the presence of IgA in human skin . In order to demonstrate the occurrence of IgA in sebaceous and sweat glands, biopsies of normal human skin were investigated and compared to intestinal mucosa . Two different commercially available anti-IgA antibodies were used . For light microscopy peroxidase-anti-peroxidase (PAP) or avidin-biotin complex (ABC) staining was used, and for electron microscopy protein-A-gold (PAG) labeling was performed on tissue sections . Specifically decorated IgA was found in sebaceous glands as well as in various portions of eccrine glands . In sebaceous glands, the maximum of IgA concentration was seen near the mouth of pilosebaceous ducts . Sweat ducts exhibited a continuous coat of IgA, whereas secretory portions contained only singular scattered IgA positive cells . Immunoelectron microscopy suggests endocytotic uptake and processing of IgA in the glandular cells . These results indicate strongly that IgA are secreted by normal human sebaceous and sweat glands . Because it is well known that IgA plays an important role in inactivation of invading viruses, bacteria, and other antigenic structures on mucous membranes, it appears that IgA in sebum and sweat fulfil a similar function on the outer body surface. Stomatologia (Athenai), 1989 Jan-Feb, 46(1), 13 - 21 {Descriptions and comparative changes of modifications on ultrastructure of cocci and bacteridium of saliva under "in vitro" influence of sparse and dense solutions of chlorhexidine di-gluconate}; Emmanouel-Nikoloussi E et al.; In our investigation we have studied and analysed the modifications on the ultrastructure of the cocci and bacteridium of saliva under "in vitro" influence of sparse and dense solutions chlorhexidine di-gluconate . We noticed that the sparse solutions of the drug revealed ruptures of the cellular wall and of the cellular membrane as well as "plasmoptysis" and "plasmolysis" . The protoplasm, showing a general coagulation, damage of the cytoplasmic membrane and the cellular wall and in concentrations of 1% and 2% we observed a disorganisation of the nuclear chromatin. Dysphagia, 1989, 3(4), 179 - 83 Pulmonary consequences of aspiration; Terry PB et al.; Aspiration can lead to serious pulmonary disease and occasionally death . Substances aspirated commonly include bacteria or gastric contents or both, but may be as unusual as diesel oil or a variety of foreign bodies . Pulmonary symptoms range from a subtle cough, wheezing, or hoarseness to severe dyspnea or asphyxiation . We discuss the mechanism of pulmonary disease caused by aspiration as well as the appropriate treatment. Oper Dent, 1989 Autumn, 14(4), 186 - 92 Histological study of an acid red caries-disclosing dye; Boston DW et al.; When 20 extracted teeth were examined histologically after using a 1.0% acid red caries-disclosing dye before excavation of carious dentin, 25% showed the presence of bacteria in the dentin . The use of a solution of 1.0% acid red in propylene glycol to identify infected dentin will greatly decrease but not completely eliminate the changes of viable bacteria remaining in a cavity preparation. Khirurgiia (Sofiia), 1989, 42(6), 87 - 8 {Peritonitis in patients treated by continuous ambulatory peritoneal dialysis}; Georgiev M et al.; With the present study the authors set themselves the task to compare the number of peritonitis episodes in patients treated with two types of systems: "Sorin-Biomedica" and "Travenol-(UV-XD)", in which disinfection of the connecting devices is achieved accordingly with chemical agents and with ultraviolet irradiation . Eleven patients have been observed from August 1984 through February 1989 . The total duration of treatment was 156 months . Twenty one peritonitis episodes were observed--15 with "Sorin-Biomedica" system and 6 with "Travenol-(UV-XD)" system--an average of one episode in 4 1/2 months with the former system and one episode in 14.7 months with the latter . It is pointed out in conclusion that the "Travenol-(UV-XD)" system with ultraviolet disinfection has significantly reduced the incidence of peritonitis at the dialysis center where the authors work. Int J Tissue React, 1989, 11(4), 165 - 8 Short chain fatty acid-induced colitis in mice; McCafferty DM et al.; Short-chain fatty acids (e.g . acetic, propionic and butyric acid) are metabolic products produced by colonic bacteria . Since intra-rectal instillation of dilute acetic acid to rats and cats has been reported to produce a diffuse colitis similar to the human disease, in our study we have investigated the effect of intra-rectal butyric acid in mice . Male BKA mice (n = 6) received 01 ml of dilute butyric acid by intra-rectal instillation which was flushed out with saline after 10 sec . Control animals received saline alone . The severity of observed inflammatory responses was assessed using an arbitrary scoring system based on clinical, macroscopic and histological features of colitis . Colitic oedema was measured as percentage (%) tissue water compared with that in saline controls . The peak oedema response was found to be at around 4 hours . A concentration-dependent symptomatic response was obtained at concentrations between 1% and 12% of butyric acid . Oedema production was similar at all concentrations above 1% . The 3% butyric acid response produced a moderate colitis with mild erythema, oedema, crypt abscess formation, goblet cell depletion and cellular infiltration without total loss of mucosal architecture . The colitogenic action of butyric acid in mouse colon could not be reproduced by low pH alone, nor by the butyrate anion at neutral or alkaline pH . These results indicate that intra-rectal butyric acid induces a consistent and reproducible colitis in mice. Ann Ist Super Sanita, 1989, 25(4), 557 - 61 Molecular approaches to the study of chemical mutagenesis; Palombo F et al.; The use of shuttle vectors has been applied in recent years to develop a better understanding of the molecular mechanisms of mutagenesis in mammalian cells . These recombinant DNA molecules replicate together with the host eukaryotic cells and can be retrieved in bacteria for rapid detection and analysis of mutation . Two approaches based on the use of shuttle vectors for studying the mutagenic effects of DNA lesions induced by alkylating agents are presented. Eye, 1989, 3 ( Pt 6), 803 - 10 'Sterile' corneal infiltrates in contact lens wearers; Bates AK et al.; Ninety four patients with 'sterile' keratitis presenting consecutively over a nine month period to the Accident and Emergency Department of Moorfields Eye Hospital were studied . This condition was found to account for 0.49% of all new casualties . A significant association was found in these patients, compared with controls, with contact lens hygiene, particularly for daily wear soft contact lenses, and contact lens case contamination by bacteria suggesting that these may be important factors in the aetiology of 'sterile' keratitis . Compared to gas permeable hard contact lenses the relative risk of developing 'sterile' keratitis in our patients was found to be 2.3 times higher with extended wear soft contact lenses, 1.56 times higher with daily wear soft contact lenses and 0.509 with polymethylmethacrylate lenses (test of trend p-value less than 0.05) . The results indicate that 'sterile' corneal infiltrates are related to contact lens hygiene and in part to contact lens case contamination by bacteria and also to the type of lens worn. Acta Vet Hung, 1989, 37(1-2), 103 - 15 In vitro evaluation of protein degradability in the rumen and digestibility of undegraded protein; Veresegyhazy T et al.; A three-phase laboratory procedure suitable for predicting protein degradability in the rumen and digestibility of undegraded protein is reported . In the first phase the feed was incubated with starch and buffered rumen fluid . In the incubation mixture the viability of protease-active bacteria was checked by anaerobic culturing, whereas changes in protease activity were monitored by azocasein degradation . In the second and third phase rumen undegradable protein (UDP) was digested with pepsin and pancreatin, respectively . The measurements showed that 63.2, 5.2 and 4.7% of the crude protein of green lucerne was decomposed by rumen fluid, pepsin and pancreatin, respectively . Degradability of the crude protein of extracted sunflower meal was 68.3, 17.7 and 5.5% in the three phases, respectively . Repeated determination yielded crude protein degradabilities of 66.7, 27.1 and 5.1% for the three phases, respectively. Tandlaegebladet, 1989 Jan, 93(1), 1 - 6 {Methods of disinfection of water systems in dental units by water chlorination}; Fiehn NE et al.; The aim of the present study was to develop a simple disinfection method to reduce the content of bacteria in the water system of dental units to an acceptable level . The study was carried out at the Royal Dental College, Copenhagen on 250 dental units . Samples of the cooling water to the ultrasonic scalers and of the water to the water glasses were obtained from eight different units representing different parts of the school . Disinfection of the water system was carried out by addition of chlorine to the pipe water near the main water intake to the institution . The chlorination af the water was automatically regulated, and the installation was so flexible that the concentration of chlorine and the time and frequency of the chlorination could be varied . Different modes of dosage of chlorine were examined . Before chlorination the bacterial content in the water system of the units was about 10(4)-10(5) c.f.u./ml . It was found that an intermittent chlorination with 0.5-1 ppm chlorine for 10 min . every day could normally reduce the bacterial counts in the water system to about a few hundreds per ml. Padiatr Grenzgeb, 1989, 28(5), 311 - 5 {Pneumococcal infections in children: hemolytic-uremic syndrome}; Handrick W et al.; By the example of a case-report diagnostic and therapeutic features of the haemolytic-uraemic syndrome are discussed within the frame of infections by pneumococci . In case of infections by pneumococci but also of those ones caused by other bacteria or viruses neuraminidase may be set free, that on its part may lead to an enzyme-induced haemolysis and in some cases also to a damage of other cell systems . In case of an active share of the kidney a haemolytic-uraemic syndrome may be the consequence. Acta Obstet Gynecol Scand, 1989, 68(3), 217 - 20 Mobiluncus and clue cells as predictors of PID after first-trimester abortion; Larsson PG et al.; Women attending our Department for a first-trimester abortion were examined for the presence of Mobiluncus species . C . trachomatis or clue cells in vaginal discharge in an attempt to identify risk groups for development of pelvic inflammatory disease (PID) after first-trimester abortion . A correlation was found between the presence of Mobiluncus and clue cells in vaginal discharge and the incidence of PID after first-trimester abortions . These women were also older than those in whom C . trachomatis was identified . Our study indicates that screening for C . trachomatis and clue cells in vaginal discharge will identify two different risk groups for developing PID after first-trimester abortionPIP: 515 women having 1st trimester abortions by vacuum aspiration were examined for risk factors of pelvic inflammatory disease (PID), specifically Chlamydia, clue cells and Mobiluncus bacteria or motile rods . C . trachomatis was seen in 7.6%, Mobiluncus and/or motile rods in 15.2%, and both in 11 . The incidence of PID after abortion was 10.8% in women with Mobiluncus, 22% in those with Chlamydia, and 4.5% in those with neither finding, excluding those pre-treated with antibiotics . 32.8% had clue cells, of whom 11.8% developed PID . 3.2% of those without clue cells developed PID . Mobiluncus was seen in 56.9% of women with clue cells, but none of those without clue cells . Thus, presence of clue cells constitutes a risk group, much easier to identify than by the tedious task of isolating Mobiluncus bacteria . Arch Otorhinolaryngol, 1989, 246(4), 218 - 21 Hemophilus influenzae adherence to and absorption from the nasal mucosa of guinea pigs; Harada T et al.; The adherence to and the absorption from nasal mucosal epithelium of type b and non-typeable strains of Hemophilus influenzae were studied in guinea pigs . H . influenzae bacteria were administered to the nose as 4 x 10(7) organisms in 1.0 ml saline . The outer membrane vesicles (OMV) of H . influenzae were used in the absorption study . Scanning electron microscopic studies showed that H . influenzae cells attached to both ciliated and non-ciliated cells of the nasal mucosa . Histological studies showed that OMV were absorbed through the nasal mucosa, mainly from ciliated cells . Further, penetration of OMV through the nasal mucosa of immunized animals was reduced when compared with that of non-immunized ones. Acta Physiol Hung, 1989, 73(2-3), 363 - 9 Intestinal post-ischemic reperfusion injury: studies with neonatal necrotizing enterocolitis; Cassutto BH et al.; In the feline intestine studies have implicated superoxide (O.-) and other oxygen derived free radicals as initiators of injury as measured by increased capillary permeability during the reperfusion period . Biochemical mechanisms of this free radical generation include: xanthine oxidase dependent O.- production, hydrogen peroxide (H2O2) formation by superoxide dismutase (SOD), hydroxyl radical (OH-) production via the Haber-Weiss reaction, and lipid radical formation from membrane peroxidation . Pathological consequences of these events include inflammatory neutrophil infiltration, damage to the collagen and mucosal basement membrane, increased capillary permeability, edema, cell degeneration and necrosis . Animal models of neonatal necrotizing enterocolitis (NNEC) indicate that intestinal injury occurs after the etiologic factors (hypothermia, hypoxia) are removed . In order to determine the role of active oxygen species in the pathogenesis of NNEC, weanling hamsters and neonatal piglets were cold stressed and activities of pro/antioxidant enzymes were determined, and histopathologic and ultrastructural studies were performed . Cold stressed weanling hamsters showed a 55.7% (P less than 0.05) decrease in xanthine dehydrogenase/xanthine oxidase activity ratio . Light microscopy revealed scattered colonic mucosal erosions and submucosal edema in 50% of cold stressed animals . Transmission electron microscopy demonstrated degeneration of colonic mucosal epithelial cells, enlarged intracellular spaces, cytoplasmic vacuolization, and nuclear membrane swelling . The colonic serosa was also edematous and infiltrated with bacteria . Large intestinal tissue from cold stressed neonatal piglets showed a significant increase (P less than 0.05) in Mn and Cu, Zn, SOD, CAT, GSH-Red, total GSH, and Glc6-PD at 0 and 12 hrs . post stress.(ABSTRACT TRUNCATED AT 250 WORDS) Int Arch Allergy Appl Immunol, 1989, 90(3), 207 - 12 Polymorphonuclear leukocyte stimulation measured by phage inactivation; Ferrini U et al.; A quantitative assay for determining the polymorphonuclear neutrophil (PMN) stimulation is described . The method is based on the inactivation of lambda vector phages that occurs after a brief exposure to stimulated PMNs . The determination of the number of residual plaque-forming units on the appropriate bacterial host allows a reproducible and sensitive quantitative assay for measuring the stimulation level of the PMN . In comparison with other methods that employ bacteria or eukaryotic cells, this assay provides several advantages and can be used for investigating the biochemical and physiological processes responsible for PMN stimulation. Geogr Med Suppl, 1989, 3, 93 - 5 {Data from measuring air pathogens of inhabitants of refuse disposal sites in Cairo (1)}; Sixl W et al.; Atmospheric germ measurements were carried out and the germ spectrum determined in medical ambulances, bedrooms and livingrooms. Geogr Med Suppl, 1989, 3, 103 - 12 Ocular infection investigations in Cairo's waste disposal sites . PVP-iodine treatment trials; Schabus V et al.; Investigations were carried out in Cairo's waste disposal sites of the inhabitants of all age groups . Germ-spectrum was analyzed before and after PVP-Iodine-treatment . However, any further dilution of PVP-Iodine in the treatment had no effect on the germspectrum, as hydrochloric acid formation in air, produced by plastic burning causes severe eye irritation and a constant lacrimation . The high germ count of the air in the dwellings and the environment is an additional source of contamination to the conjunctiva. Adv Perit Dial, 1989, 5, 227 - 8 Comparison of two non-disconnect CAPD delivery systems; Hall LJ et al.; Intraluminal transmission of bacteria remains a significant factor in the morbidity and procedural success of CAPD . A worsening rate of peritonitis in a longstanding CAPD program (first patients 1978) led to a search for a system which might allow a lesser rate of peritonitis . Simplicity in the procedure was a requirement . For this reason disconnect systems were not considered . The Travenol spike system was prospectively compared with Delmed leur lock in terms of rate of peritonitis and difficulty in training . The former had been used by the center since inception of the CAPD program . 28 patients new to CAPD were alternately assigned to each system without other bias, including diagnosis, age, sex, or race . The study, while ongoing, was analyzed at 12 months . 66 patient months were involved with each system . The peritonitis rates were: 2.2 episodes: patient year with the Travenol; 0.9 episode:patient year for the Delmed systems . The Delmed system appears to provide a lower rate of peritonitis . In addition, less manual dexterity, steadiness, and hand eye coordination are necessary. Int J Fertil, 1989, 34 Suppl, 85 - 7 Noncontraceptive health benefits and risks of steroidal contraception; Pasquale SA; PIP: From the extensive research conducted over the past 28 years, there is a clear picture that the noncontraceptive benefits of steroidal contraceptives are considerable and the benefits outweigh the risks . The risks associated with the increased incidence of thromboembolic disease have reduced with lower doses of both estrogen and progesterone . Also, the increased risk of hepatocellular carcinoma is very low, compared with the benefits . One benefit is the reduction in primary dysmenorrhea which was discovered in 1940 . This occurs due to the suppression of ovulation and decrease in endometrial growth . Ovarian cysts resolve spontaneously; 3500 fewer hospitalizations due to ovarian cysts are reported for 1982 . 11,000 fewer cases of ectopic pregnancy/year are a result of oral contraceptive (OC) use . Retrospective case studies have found that pelvic inflammatory disease (PID) is prevented by use of OCs . This happens because the cervical mucus remains thick throughout the menstrual cycle with OC use, and thus prevents transportation of bacteria by sperm from the lower to the upper genital tract . Another reason is the decreased amount of blood flow at the time of withdrawal provides a less conducive environment for bacteria growth . 15,000 annual hospitalizations for PID are estimated to have been prevented by OC use . The data on breast cancer are conflicting, but most do not show a link between OCs and breast cancer . In fact, benign breast disease may be reduced by 23,000 annual hospitalizations due to OC use . Another benefit of OC use is the decreased incidence of endometrial and ovarian cancer . The relative risk among OC users in 1987 was estimated at P = 0.6 for primary endometrial cancer . This beneficial effect continues after OC use is discontinued . There is a 40% reduction in the incidence of ovarian cancer among OC users compared with nonusers, and is related to duration of use, but the protective effect continues after OC use discontinuation . Bone mass is increased in women who use OCs, although further study is required to determine whether the increased bone mass protects from osteoporosis after menopause . Arch Histol Cytol, 1989, 52 Suppl, 277 - 92 Phylogenetic considerations of neurosecretory granule contents: role of nucleotides and basic hormone/transmitter packaging mechanisms; Payne CM; The characteristics of neurosecretory granules include the presence of an acidic interior, a hyperosmolar concentration of granule solutes, the presence of chromogranin (CG) or CG-like soluble acidic proteins and a high content of nucleotides, predominantly ATP . The identification of "nucleotides" within the neuroendocrine "stem cells" of coelenterates (e.g . Hydra) has raised some interesting evolutionary questions as to the function of intragranular nucleotides . The chromaffin granules of adrenal medullary cells have been studied extensively, and are representative of the neurohormone/neurotransmitter packaging problems encountered in neurosecretory granules, in general . At the acid pH (5.7) of the interior of the chromaffin granule, ATP has three negative charges based on the pK value of the gamma-phosphate group . ATP can therefore interact with positively charged amines, acetylcholine and divalent cations, forming binary and ternary complexes . The results of nuclear magnetic resonance (NMR) spectroscopy indicate that the hyperosmolar solutes within the chromaffin granule exist in a viscous, but fluid state; one function of ATP could be to help lower the osmotic pressure of the granule contents through extensive, but weak, intermolecular bonding . In addition, ATP is an excellent buffer to help maintain a pH of 5.7 within the interior of the chromaffin granule . An acidic milieu contributes to neurohormone/neurotransmitter packaging and granule stability . The presence of nucleotides within neurosecretory granules cannot, however, be explained on the basis of the ability of ATP to simply reduce osmotic pressure, since insulin molecules exist in a crystalline phase, a condition which, by itself, could substantially reduce osmotic pressure; nucleotides, nevertheless, co-exist in these insulin cores . ATP and ATP metabolites such as ADP, AMP and adenosine, formed as a result of the action of ectonucleotidases, can have extensive extracellular trophic and feedback effects after secretion . Extracellular nucleotides and adenosine can function as neuromodulators, agonists and antagonists to inflammatory cells, and regulators of blood flow, etc . It is possible that intragranular nucleotides were retained through a billion or more years of evolution because of the importance of these trophic and feedback effects . Parts of the neuroscretory granule, such as the F1 subunit of the proton-translocating ATPase, can be traced back to the aerobic bacteria, vacuolar amine transport to yeast and a CG-like acidic protein to protozoan secretory granules (i.e., the trichocysts of Paramecia). Arch Neurobiol (Madr), 1989, 52 Suppl 1, 30 - 44 {Neuropathology of HIV infection}; Gutierrez Molina M et al.; Some references about the important neuropathologic lesions observed in patients with AIDS and about the variation on the relative incidence of the processes detected when analyzing clinic and neuropathologic tables, as well as a mention to the confusion created by terminological problems, clinicopathologic disagreement and wrong etiologic attributions, are used as an introduction to the exposition of HIV infection neuropathology . In a first section, Central Neuropathology is described, starting with the disorders considered as primarily caused by HIV . Then, infectious secondary Neuropathology, caused by different viruses, parasites, fungi or bacteria, and neoplastic secondary Neuropathology, mainly represented by localized lymphoma and by the uncommon Kaposi's disease, are analyzed . This section also includes references to cerebrovascular complications, Pediatric Neuropathology and ends with an analysis of cerebral biopsy indications . In the second part, Peripheral Neuropathology is described, splitting up peripheral nerve and muscle pathology. Digestion, 1989, 44(2), 95 - 100 Interdigestive gastroduodenal motility in patients with active and inactive duodenal ulcer disease; Bortolotti M et al.; The interdigestive gastroduodenal motility was studied by means of a manometric probe in 6 patients with active duodenal ulcer and acid hypersecretion, in 6 patients with ulcer disease in remission (inactive) and normosecretion and in 8 healthy subjects with normosecretion . After a basal recording period sufficient to record at least two activity fronts of the migrating motor complex (MMC), an intraluminal infusion of isotonic NaHCO3 was carried out for 180 min in patients with active duodenal ulcer, whereas in patients with ulcer in remission an HCl solution was infused for 180 min . Patients with active duodenal ulcer showed a basal motility with a longer than normal MMC cycle and a shorter than normal activity front, while patients with ulcer in remission showed a cyclic motor activity not significantly different from that of normal subjects . The NaHCO3 infusion in patients with active ulcer restored a near-normal motility, whereas the HCl infusion in patients with ulcer in remission induced a motility similar to that of patients with active ulcer . These data indicate that the increase in gastric acid secretion is responsible for the decrease in frequency and duration of MMC activity fronts, which have the function of cyclically clearing the gastroduodenal lumen . Consequently, acid and bacteria may remain a longer than normal time in contact with the gastroduodenal mucosa, which, in this manner, may be greatly exposed to the risk of peptic lesions. Int J Biochem, 1989, 21(6), 657 - 60 Adenylate kinase of Mycobacterium marinum . Amino acid composition, secondary structure and other aspects; Batra PP et al.; 1 . Adenylate kinase purified from Mycobacterium marinum shares homology with the enzyme from mammalian muscle and from other bacteria with respect to amino acid composition, total residues and Mr . 2 . However, it differs from others in not containing a free sulfhydryl or disulfide group . 3 . It is much more sensitive to inhibition by adenosine (5')pentaphospho(5')adenosine than is the enzyme from other sources . 4 . The enzyme contains 24% alpha-helix, 30% beta-structure and 46% random coil. Pediatriia, 1989, (7), 32 - 5 {Clinico-laboratory analysis of the diarrhea syndrome in young children excreting rotaviruses with feces}; Nikovskaia MI et al.; A total of 404 children admitted to the infectious hospital of the non-intestinal type were examined with the aid of counter immunoelectroosmophoresis . Rotavirus antigen was detected in feces in 46.8% of 126 children admitted with a diagnosis of diarrhea, in 10.8% of 278 without diarrhea, and also in 32.9% of 149 children with diarrhea that occurred at the hospital . The clinical course of acute rotavirus gastroenteritis appeared to be materially influenced by acute respiratory viral infection, particularly influenza, and by certain types of opportunistic bacteria contained in high concentrations by the intestine. Crit Rev Biochem Mol Biol, 1989, 24(4), 271 - 328 Leukocytic oxygen activation and microbicidal oxidative toxins; Hurst JK et al.; Following a brief introduction of cellular response to stimulation comprising leukocyte activation, three major areas are discussed: (1) the neutrophil oxidase; (2) myeloperoxidase (MPO)-dependent oxidative microbicidal reactions; and (3) MPO-independent oxidative reactions . Topics included in section (A) are current views on the activation mechanism, redox composition, structural and topographic organization of the oxidase, and its respiratory products . In section (B), emphasis is placed on recent research on cidal mechanisms of HOCl, including the oxidative biochemistry of active chlorine compounds, identification of sites of lesions in bacteria, and attendant metabolic consequences . In section (C), we review the (bio)chemistry of H2O2 and .OH microbicidal reactions, with particular attention being given to addressing the controversial issue of probe methods to identify .OH radical and critical assessment of the recent proposal that MPO-independent killing arises from site-specific metal-catalyzed Fenton-type chemistry. Life Sci, 1989, 44(26), 2057 - 65 Adrenergic receptor homologies in vertebrate and invertebrate species examined by DNA hybridization; Palacios JM et al.; The deduced protein sequences of the mammalian adrenergic receptors (ARs) suggest that these proteins have evolved by several ancient gene duplication events . To investigate in what species these events may have occurred DNA fragments encoding the family of adrenergic receptors from human (beta 1AR and alpha 2AR) and hamster (beta 2AR and alpha 1AR) were used to detect homologous sequences in other vertebrates, invertebrates and unicellular organisms by Southern blot hybridization analysis . Sequences homologous to hamster beta 2AR were detected in lower vertebrates, invertebrates and Dictyostelium, but not in yeast or bacteria . Within vertebrates, sequences strongly homologous to human beta 1AR and human platelet alpha 2AR were confined to the higher vertebrates only . In the invertebrates, only Drosophila contained sequences homologous to hamster alpha 1AR . Our results suggest that non-mammalian species may contain receptors homologous to the mammalian adrenergic receptors and that the sequences homologous to human beta 2AR have been the most strongly conserved. Geogr Med Suppl, 1989, 2, 39 - 60 {Epidemiologic significance of sludge for the transmission of zoonoses}; Kock M; Sludge contains all types of pathogenic organisms originating from the drainage system of the population . Decontamination is difficult and expensive, it is ranging from thermal-chemical to radioactive treatment of the materials . Suitable treatment and use of sludge can really reduce the health hazards to the population. Biochem J, 1989 Jan 1, 257(1), 87 - 94 Studies on electron transfer from methanol dehydrogenase to cytochrome cL, both purified from Hyphomicrobium X; Dijkstra M et al.; Ferricytochrome cL isolated from Hyphomicrobium X is an electron acceptor in assays for homologous methanol dehydrogenase (MDH), albeit a poor one compared with artificial dyes . The intermediates of MDH seen during the reaction are identical with those observed with Wurster's Blue as electron acceptor, indicating that the reaction cycles are similar . The assay showed a pH optimum of approx . 7.0 and scarcely any stimulation by NH4Cl, this being in contrast with assays with artificial dyes, where strong activation by NH4Cl and much higher pH optima have been reported . From the results obtained with stopped-flow as well as steady-state kinetics, combined with the isotope effects found for C2H3OH, it appeared that the dissimilarities between the electron acceptors can be explained from different rate-limiting steps in the reaction cycles . Ferricytochrome cL is an excellent oxidant of the reduced MDH forms at pH 7.0, but the substrate oxidation step is very slow and the activation by NH4Cl is very poor at this pH . At pH 9.0 the reverse situation exists: ferricytochrome cL is a poor oxidant of the reduced forms of MDH at this pH . No C2H3OH isotope effect was observed under these conditions, indicating that substrate oxidation is not rate-limiting, so that activation by NH4Cl cannot be found . Since just the opposite holds for assays with artificial dyes, the poor electron-acceptor capability and the different pH optimum of ferricytochrome cL as well as the insignificant activating effect of NH4Cl (all compared with artificial assays) can be explained . Although different views have been reported on the rate-limiting steps in the systems from Methylophilus methylotrophus and Methylobacterium sp . strain AM1, these are most probably incorrect, as rate-limiting electron transfer between ferrocytochrome cL and horse heart ferricytochrome c can occur . Therefore the conclusions derived for the Hyphomicrobium X system might also apply to the systems from other methylotrophic bacteria . Comparison of the assays performed in vitro (at pH 7.0) having ferricytochrome cL and Wurster's Blue as electron acceptor with methanol oxidation by whole cells shows that the former has similarity whereas the latter has not, this being although ferricytochrome cL is a poor electron acceptor in the assay performed in vitro . The reason for this is the absence of a (natural) activator able to activate the (rate-limiting) substrate oxidation step at physiological pH values. J Pharm Biomed Anal, 1989, 7(2), 227 - 32 Quality assessment of DNA vaccines: hepatitis-B vaccine; Peetermans JH; Vaccines manufactured following "classical" methods contain inactivated or infectious but attenuated viruses or bacteria . In some instances, the inactivated agents are purified . In other cases, the vaccines contain protein subunits or practically pure polysaccharides . It is generally accepted that the final product cannot be completely characterised and that therefore extensive "in-process" controls are necessary to prove the consistent quality of such vaccines . Control tests are carried out on the substrate, the pooled bulk vaccine and on the final containers . Vaccines produced by recombinant DNA techniques consist of pure proteins . The production is carried out by the multiplication of the "working seed" under well-defined standardised conditions followed by clarification, extraction, purification, formulation . "In-process" controls are incorporated at each step and specifications for acceptance are formulated . The biological methods used for the classical vaccines are completed by physicochemical and immunological determinations of antigen content, identity and purity for the "new generation" products . The requirements for the manufacturers are based on the documents issued by the World Health Organisation and by the national control authorities . The marketing of vaccines is based on a lot by lot release procedure, whereby each lot is tested by the manufacturer and the national control authority before use . Hepatitis-B vaccine, derived from transformed yeast cells, is the first and sole vaccine which has obtained a world-wide license . The quality assessment of this vaccine has been achieved following the requirements for the new generation of biomolecules . It is an example for future vaccines. Arch Exp Veterinarmed, 1989, 43(5), 755 - 61 {Natural and experimental infections of lambs with Mycoplasma ovipneumoniae}; Bocklisch H et al.; Mycoplasma (M.) ovipneumoniae was isolated pure or mixed with bacteria from 47 lungs of lambs of 14 in 22 tested flocks . M . ovipneumoniae was obtained as pure culture in cases of mild bronchopneumonia . Experimental intratracheal or intranasal infection caused several days of rising body temperature above 39.7 degrees C . Nasal discharge, coughing, and dyspnea did not occur . M . ovipneumoniae was successfully re-isolated from nasal swabs, beginning 2 d from infection . Lobular catarrhal bronchopneumonia was established by postmortem examinations, 10-14 d from infection, and M . ovipneumoniae was re-isolated from the lungs . Histological patterns of lungs were characterised by interstitial cell reactions. J Mol Cell Immunol, 1989, 4(4), 191 - 9; discussion 199-201 Regulation of Fc gamma 2a receptor-mediated phagocytosis by a murine macrophage-like cell line, P388D1: involvement of casein kinase II activity associated with Fc gamma 2a receptor; Yamada A et al.; A question of whether or not casein kinase II (CKII) activity associated with Fc gamma 2aR is involved in the regulation of phagocytic process mediated by this type of Fc gamma R was investigated . Our previous studies showed that the rate of phagocytosis of sheep erythrocytes (SRBC) coated with anti-SRBC antibody (EA) by P388D1 cells varies significantly depending on the isotypes of antibody and that Fc gamma 2aR isolated from the detergent lysate of P388D1 cells is associated with CKII activity, whereas Fc gamma 2bR is not . Fc gamma Rs-mediated phagocytosis is a major function of macrophages by which invading pathogens such as bacteria could be eliminated and therefore warrants the investigation of its biochemical mechanisms . We have recently shown that phagocytosis of EA2b mediated by Fc gamma 2bR of P388D1 cells as well as murine peritoneal macrophages could be up-regulated by promoting the association of various cytoskeletal components with the receptor by inhibiting Fc gamma 2bR-associated phospholipase A2 (PLA2) . CKII activity-associated Fc gamma 2aR mediates phagocytosis of EA2a more effectively than PLA2-associated Fc gamma 2bR mediates phagocytosis of EA2b . We have therefore examined a potential role of CKII in Fc gamma 2aR-mediated phagocytosis by the use of a specific inhibitor of CKII activity (heparin) . Results showed that heparin inhibited CKII activity associated with Fc gamma 2aR and effectively down-regulated the Fc gamma 2aR-mediated phagocytosis by apparently blocking the association of the receptor with four types of cytoskeletal components (actin-binding protein, myosin heavy chain, alpha tubulin, and actin).(ABSTRACT TRUNCATED AT 250 WORDS) G Ital Dermatol Venereol, 1989 Jan-Feb, 124(1-2), 17 - 9 {Seborrheic keratoses studied using scanning electron microscopy}; Bencini PL et al.; Ten seborrheic keratosis of acanthotic type have been observed by S.E.M . The morphological study could confirm the existence of two processes in the origin of the horny cysts which are peculiar to this affection . a) an extrafollicular process causes the involvement of groups of keratinocytes which, by assuming a vortex disposition, produce a concentric keratinization; b) a follicular process which causes an accentuated keratinization of the cells surrounding the epithelial sheath . This process was associated to the presence of multiple hairs a single follicle . Moreover, with higher magnification, it was possible to observe a great number of bacteria, yeasts and of particular alteration of the membrane which could be compared to changes in the tone filaments. Zahn Mund Kieferheilkd Zentralbl, 1989, 77(1), 12 - 6 {The concentration and duration of the action of metronidazole in the gingival pocket following local application}; Passler R et al.; Periodontitis is a locally limited disease caused by bacteria . The local application presents itself useful for the indicated medicamentous therapy of periodontitis by means of metronidazole . In preliminary tests the metronidazole liberation from hollow fibres, in gel and from polyvinyl alcohol platelets has been tested by ultraviolet absorption measurement . The hollow fibres have been evaluated no more because of too small agent absorption in the clinical test . In polyvinyl alcohol metronidazole acts definitely over 3 days . However, the prolonged action in the gel form is not definitely (45 min to 24 h) . A single one or repeated twice metronidazole/polyvinyl alcohol platelet application results in a sufficient long and high metronidazole concentration in the gingival pocket. Nephron, 1989, 52(1), 29 - 35 Exaggerated glomerular albuminuria after cobra venom factor in anti-glomerular basement membrane disease; Savige JA et al.; The mechanisms by which intercurrent bacterial infections are associated with increased tissue injury in some forms of glomerulonephritis may include complement activation by bacteria and subsequent increased glomerular neutrophil (PMN) infiltration . We have studied the effect of complement activation after cobra venom factor (CVF) in anti-glomerular basement membrane (GBM) antibody-mediated disease . A single injection of CVF 24 h before the administration of heterologous nephrotoxic globulin (NTG) to Sprague-Dawley rats resulted in greatly increased albuminuria in some animals on the second day of this model . This phenomenon was reproducible and depended on the presence of circulating PMN and complement . We have previously shown that the administration of CVF on days 9 and 11 of the HgCl2 model in inbred Brown Norway rats, resulted in increased albuminuria in all animals at day 17 (p less than 0.05) . The administration of small amounts of CVF with consequent complement activation in antibody-mediated disease represents a model for the increased injury seen after infection in human disease. J Assoc Acad Minor Phys, 1989, 1(1), 24 - 9 Management of severe acute pancreatitis: an evolving approach; Debas HT; No major breakthrough has occurred in our understanding of the etiology or pathogenesis of acute pancreatitis . However, significant advances in management of the disease have resulted in improved survival . These include disease severity assessment by Ranson's criteria or by similar other clinical methods, as well as by computerized tomography (CT) . The use of contrast-enhanced CT has made possible early diagnosis of pancreatic necrosis by showing areas of hypoperfusion . Once the presence of necrosis is established, CT-guided needle aspiration of necrotic tissue can be performed for gram stain and for culture of bacteria . This approach makes possible earlier diagnosis of pancreatic sepsis, and hence earlier surgery . Other important contributions have been radiologic and endoscopic interventional techniques . Percutaneous catheter drainage of rapidly enlarging acute pseudocysts can obviate the serious complication of free rupture of pseudocysts into the peritoneal cavity; similar drainage of infected pseudocysts may also provide a definitive or at least a palliative therapy . Bleeding from eroded vessels, false aneurysms or pseudocysts can often be successfully controlled by selective angiography . In severe acute biliary pancreatitis, endoscopic sphincterotomy within 72 hours of admission has been shown to reduce morbidity, mortality, and hospital stay . These interventional techniques permit surgery to be postponed until the most optimal time . The advances cited above, as well as improved intensive care and nutritional management, are beginning to reduce the high mortality of severe acute pancreatitis. Comp Biochem Physiol B, 1989, 94(4), 759 - 63 Biochemical identification of thymosin alpha-1: its phylogenetic distribution and evolutionary implications; Oates KK et al.; 1 . Thymosin alpha-1, like reactivity, was found in several different species (insects, crab, protozoan, fungus and bacteria) by radioimmunoassay and immune fluorescence and as an extracellular product from the bacterial genus Mycobacterium . 2 . Biochemically, thymosin alpha-1 has been isolated from combined crab visceral and nervous tissue by reverse phase HPLC . 3 . The identification of thymosin alpha-1 in lower life forms suggests a more generalized exocrine origin in unicellular organisms prior to the development of the immune system or exocrine differentiation. Gerontology, 1989, 35(2-3), 127 - 9 Effect of age on adherence of Branhamella catarrhalis to buccal epithelial cells; Carr B et al.; The role of adherence in the pathogenicity of Branhamella catarrhalis in lower respiratory tract infection in the elderly was investigated . Differential adherence of this organism to buccal epithelial cells of elderly inpatients, outpatients, and young controls was measured . The mean number of bacteria adherent per cell was 36.9 +/- 11.2, 40.2 +/- 6.1 and 16.5 +/- 12.8 for inpatient and outpatient elderly and young controls respectively (significantly different p less than 0.01, Student's t test) . This significantly higher level of adherence in the elderly subjects suggests that increased adherence may contribute to the incidence of respiratory tract infection due to B . catarrhalis seen in this group. Arch Microbiol, 1989, 152(2), 154 - 9 Molecular analysis of methane monooxygenase from Methylococcus capsulatus (Bath); Stainthorpe AC et al.; Methane monooxygenase (MMO) is the enzyme responsible for the conversion of methane to methanol in methanotrophic bacteria . In addition, this enzyme complex oxidizes a wide range of aliphatic and aromatic compounds in a number of potentially useful biotransformations . In this study, we have used biochemical data obtained from purification and characterization of the soluble MMO from Methylococcus capsulatus (Bath), to identify structural genes encoding this enzyme by oligonucleotide probing . The genes encoding the beta and gamma subunits of MMO were found to be chromosomally located and were linked in this organism . We report here on the analysis of a recombinant plasmid containing 12 kilobases of Methylococcus DNA and provide the first evidence for the localization and linkage of genes encoding the methane monooxygenase enzyme complex . DNA sequence analysis suggests that the primary structures of the beta and gamma subunits of MMO are completely novel and the complete sequence of these genes is presented. Acta Leprol, 1989, 7 Suppl 1, 85 - 8 Rapid and precise diagnosis of atypical mycobacterial infection by chemotaxonomical and immunological methods; Ikawa H et al.; The species of 205 strains of acid fast bacteria isolated from swine and human mycobacteriosis were identified chemotaxonomically and numericaltaxonomically . The species of the isolates which were identified numericaltaxonomically as Mycobacterium avium intracellulare (MAI) complex were further classified by using both thin-layer chromatography of the antigenic glycopeptidolipids (GPL) from the bacteria and seroagglutination test devised by Schaefer . These MAI complex from swine fell into serotype 8 (45 strains), serotype 4 (32 strains), serotype 9 (9 strains) and untypable (9 strains), respectively . In contrast to swine, human isolates covered more wide ranges of serotypes such as serovar 7, 12, 16 besides serovar 4, 8 and 9 . Furthermore, enzyme-linked immunosorbent assay (ELISA) which is based on the type specific glycolipid antigen and infected swine/human sera was applied to distinguish serological variants of the MAI complex . Of the fourteen cases in swine and five in human that had been typed by both the seroagglutination reaction and the thin layer chromatography (TLC) the thirteen in swine and two in human cases showed clear coincidence with the results of ELISA . The results demonstrated that enzyme-linked immunosorbent assay using infected sera was especially useful, and it was recommended from the sensitivity and rapidity as an adjunct to seroagglutination test and thin layer chromatography for the identification of serotypes of MAI complex. Acta Leprol, 1989, 7 Suppl 1, 77 - 80 Methionine as methyl-group donor in the synthesis of Mycobacterium avium envelope lipids, and its inhibition by DL-ethionine, D-norleucine and DL-norleucine; David HL et al.; The radioactivity from 3H-methyl methionine was rapidly incorporated into the surface lipids of Mycobacterium avium . The transmethylation reaction was efficiently inhibited by DL-ethionine, D-norleucine and DL-norleucine . The structure of the outerlayer of the M . avium envelope was profoundly altered in the bacteria treated with DL-norleucine. Acta Leprol, 1989, 7 Suppl 1, 173 - 4 Phagosome-lysosome fusions in macrophages infected by Mycobacterium avium: role of mycosides-C and other cells surface components; Frehel C et al.; The phagosome-lysosome fusions (PLE) were assessed in case of bone-marrow macrophages infected by the opportunistic species Mycobacterium avium, employing the acid-phosphatase (AcPase) electron-cytochemistry . The role of surface components was evaluated by coating the bacteria prior to phagocytosis by specific M . avium antiserum or the anti-mycosides-C serum raised in rabbit . PLF was evaluated under the electron microscope during (2, 4 hours), or after (24 hours) phagocytosis . The preliminary results suggest that although M . avium surface components intervene in PLF inhibition, the role of mycosides-C among these surface components (effectively intervening in PLF inhibition) is questionable. Acta Leprol, 1989, 7 Suppl 1, 160 - 3 Evidence that host-recycling of Mycobacterium avium preserves its ability to hinder macrophage killing functions; Frehel C et al.; The opportunistic pathogen Mycobacterium avium was selected as a model for the study of the bacterial cell envelope and resistance to macrophage killing functions . We hereby demonstrate that characteristic features of M . avium, e.g . existence of a polysaccharide-rich outer layer (POL), presence of a protective capsule also called electron-transparent zone (ETZ) around phagocytized bacteria, and inhibition of phagosome-lysosome fusions (PLF), were better expressed by bacteria recently isolated from infected rabbits than by bacteria subcultured in laboratory media . Our data appeared to confirm previous suggestions that M . avium regulatory mechanisms are such that during laboratory growth of these bacteria, synthesis of surface components which may be important concerning their virulence properties, is effectively diminished. Zhonghua Kou Qiang Yi Xue Za Zhi, 1989 Jan, 24(1), 1 - 3, 62 {Histochemical studies of three kinds of enzymes in dentinal caries}; Liu PX; Aminopeptidase, acid phosphatase and nonspecific esterase in dentinal caries were investigated by histochemical method in this study . It was found that in contrast with Gram's stain, the several hydrolytic enzymes were all located within the region where bacteria invaded the dentine . This suggested that the enzymes were produced by the bacteria invading dentinal tubules . According to previous study of the functions of these enzymes, it was speculated that in the process of dentinal caries, aminopeptidase and acid phosphatase may be involved in the decomposition of structural protein of dentinal organic matrix and non-specific esterase in the decomposition of structural lipids of dentine. Can J Microbiol, 1989 Jan, 35(1), 30 - 5 An archaebacterial promoter sequence assigned by RNA polymerase binding experiments; Thomm M et al.; To identify an archaebacterial promoter sequence, nuclease protection studies with the purified RNA polymerase of Methanococcus vannielii were performed . The enzyme binds specifically both at protein-encoding (hisA and methyl CoM reductase, component C) and tRNA-rRNA genes . The binding region of the RNA polymerase extends from 30 base pairs (bp) upstream (-30) to 20 bp downstream (+20) from the in vivo transcription start site . This finding indicates that the archaebacterial enzyme recognizes promoters without transacting transcription factors . The DNA segment protected from nuclease digestion by bound RNA polymerase contains an octanucleotide sequence centered at -25, which is conserved between the protein-encoding and the stable RNA genes . According to the specific binding of the enzyme to only DNA-fragments harbouring this motif, we propose the sequence TTTATATA as the major recognition signal of the Methanococcus RNA polymerase . Comparison of this motif with published archaebacterial DNA sequences revealed the presence of homologous sequences at the same location upstream of 36 genes . We therefore consider the overall consensus TTTATAATA as a general element of promoters in archaebacteria . In spite of the specific binding of the enzyme, most preparations of the Methanococcus vannielii RNA polymerase are unable to initiate transcription at the correct sites in vitro . Here we present first evidence for the possible existence of a transcription factor conferring the ability to the enzyme to initiate and terminate transcription specifically in vitro. Proc Natl Acad Sci U S A, 1989 Jan, 86(1), 138 - 41 Characterization of hydrogen-uptake activity in the hyperthermophile Pyrodictium brockii; Pihl TD et al.; Pyrodictium brockii is a hyperthermophilic archaebacterium with an optimal growth temperature of 105 degrees C . P . brockii is also a chemolithotroph, requiring H2 and CO2 for growth . We have characterized P . brockii hydrogen-uptake activity with regard to temperature, ability to couple hydrogen oxidation to artificial electron acceptor reduction, sensitivity to O2, and cellular localization . The hydrogen-uptake activity was localized predominantly in a particulate fraction, was reversibly inhibited by O2, and coupled H2 uptake to the reduction of positive potential artificial electron acceptors . Comparisons between these results and those of the well-studied hydrogen-uptake hydrogenase from the mesophile Bradyrhizobium japonicum showed the two enzymes to be similar despite the very different natural environments of the organisms . However, the optimum temperature for activity differed greatly in the two organisms . We have also used immunological and genetic probes specific to the 65-kDa subunit of B . japonicum hydrogenase to assay crude extracts and genomic DNA, respectively, from P . brockii and found the enzymes to be similar in these respects as well . In addition, we report a formulation for artificial seawater capable of sustaining the growth of P . brockii. Dakar Med, 1989, 34(1-4), 166 - 71 {Epidemiological study of diarrhea in children 0-5 yr of age}; Fall M et al.; The authors use an investigation carried out at Pikine Social Paediatric Institute on children consulting for diarrhoea to underline the epidemiological factors of this affliction in Dakar suburbs: the average age of the children is 14 months; almost half the children came from families with low or irregular incomes; coprologic examination showed parasitic germs in 26.7% of the cases, rotavirus in 24.4%, and bacteria in 17.6%. Dakar Med, 1989, 34(1-4), 134 - 8 {Epidemiological aspects of neonatal purulent meningitis in Dakar}; Faye/Ndao MA et al.; The authors point out that, after tetanus, purulent meningitis is a very common neo-natal infection . Their epidemiology is dominated by the existence of particular obstetric and associated factors, which play an important role in the early development of this sickness . Numerous forms of bacteria, particularly pneumococci, were isolated . In spite of everything, the mortality rate is high, so attention should be given to preventing this illness . Better follow-up during pregnancy coupled with better conditions for delivery could help this prophylaxis. Electron Microsc Rev, 1989, 2(2), 231 - 55 Melamine resins and their application in electron microscopy; Frosch D et al.; Melamine resins are derived from the heterocyclic compound triaminotriazine, C3H6N6 . Similar to proteins in structure and reactivity, water-soluble melamine resins can be used as water-embedding media for electron microscopy (Bachhuber and Frosch, 1983) . The idea behind this approach was to study some of the artefacts of traditional embedding techniques and to work out conditions to eliminate as far as possible denaturing of proteins and extraction of lipids . Sectioned cells and tissues processed in the melamine resin Nanoplast show remarkable preservation of ultrastructure . Because they can be sectioned extremely thinly, melamine resins are particularly suitable for dark-field and electron spectroscopic imaging of unstained molecular suspensions providing in this way an unusually clear reproduction of ultrastructural detail such as the helical structure of isolated unstained double-strand DNA molecules (Frosch et al., 1987b) . In 1988, the melamine resin Nanostrat was introduced as an EM-compatible prolific substrate foil for cell culture (Westphal et al., 1988) . Cells or bacteria cultivated on this material can be processed for various kinds of follow-up techniques like TEM, SEM, vertical sectioning and immunocytochemistry. J Conn State Dent Assoc, 1989 Summer, 63(3), 337 - 41 Infection control update: beware of the ragwheel; Noraian KW; Previously, concern about infection control in dentistry emphasized the handpiece, operator safety, barrier technique, and patient protection in the dental operatory . As current knowledge of infection control increases an expanded understanding of other sources of disease transmission is indicated . Often overlooked is the dental laboratory of which particular interest focuses on the lathes used in preparing prosthetic appliances, castings, orthodontic appliances, and surgical stents . Specifically, the potential threat of the ragwheel to the patient and operator is significant . Ragwheels and pumice samples were collected and cultured, the results of which mandate the need for infection control guidelines for the dental laboratory. Acta Biochim Biophys Hung, 1989, 24(4), 299 - 311 The effect of L-leucine methyl ester on the phagocytosis and amino acid incorporation of murine peritoneal cells; Antoni F et al.; Murine peritoneal macrophages were treated in vitro with L-leucine methyl ester (0.25-5.0 mM) . This treatment resulted in an inhibition of the amino acid incorporation into the cells both at 4 degrees C and 37 degrees C during a relatively short incubation period . The adherence of macrophages was not changed by the treatment . Bacterial phagocytosis was partly influenced: Leu-OMe did not change the binding but the engulfment of opsonized bacteria was blocked . Damage of the plasma membrane caused by Leu-OMe was not so serious as that produced by specific anti-PEC antiserum . Leu-OMe is a lysosomotropic agent accumulated preferentially by lysosomes . The vacuolization of the cells and the dilatation of the vacuoles are evidences for the intracellular damage . In the early phase this damage is characterized only by the leakage of the cytoplasm, later the damage of the plasma membrane can also be demonstrated. Front Med Biol Eng, 1989, 1(2), 89 - 97 Biomedical engineering for the conservation of teeth--the use of a Nd-YAG laser for a treatment of apical focus; Sumitomo M et al.; When the apical focus (root cyst) is treated, if the tooth concerned can be preserved, the root canal should first be enlarged and cleaned sufficiently, and then sealed with filling materials . However, if an expensive prothesis is placed on the crown of the tooth concerned, or if the root canal is blocked with a post core of calcification, root canal treatment becomes difficult from a practical point of view . We therefore tried irradiation using the Nd-YAG laser, which is known for its high transmissibility into teeth, to the root canal, and the apical area . In its histological images it is considered to the action on bacteria and bacteria-infected substances inside the root canal through calcification of the dentinal surface layer facing the dental pulp, closure of the dentinal canal opening, and there was substantial change in the dentine in the outer layer . We have achieved good clinical results using this method; we eventually extracted only two teeth in 200 cases; it is thus very significant that most of the teeth of the patients still maintain their function, although they were diagnosed as non-preservable using conventional methods. G Stomatol Ortognatodonzia, 1989, 8(3), 18 - 20 {Acidogenesis and fluoride}; Calderari G et al.; The Authors consider the inhibition of acidogenesis in the dental plaque by topical application of fluoride . After an examination of the actions of fluoride on the bacterial component of dental plaque, the Authors come to the conclusion that inhibition of acidogenesis is one of the possible anticaries mechanism of topical fluoride. Biol Met, 1989, 2(1), 31 - 5 Ferrochelatase activity in Azospirillum brasilense with reference to the influence of metal cations; Price MJ et al.; Ferrochelatase in membrane preparations from Azospirillum brasilense displayed an activity of 2.17 mumol protoheme formed.h-1.mg protein-1 which is 10-fold greater than previous reports for other bacteria . This ferrochelatase showed an apparent Km of 20.9 microM for Fe2+, a pH optimum of 6.0-6.5, and stimulation by oleic or stearic acids . Co2+, Cu2+ and Zn2+ inhibited the incorporation of Fe2+ into protoporphyrin IX while Ni2+ and Mg2+ had no effect on protoheme synthesis . Activity with Fe2+ and mesoporphyrin IX was less than with protoporphyrin IX but deuteroporphyrin IX produced the highest rate of protoheme synthesis . The membrane fraction containing ferrochelatase activity was found to insert Cu2+, Ni2+, Zn2+ and Co2+ enzymatically into protoporphyrin IX to produce metalloporphyrins . Cu2+ incorporation into protoporphyrin IX proceeded at a rate greater than with Fe2+ and the Km for Cu2+ was 21.9 microM. Aust J Biotechnol, 1989 Jan, 3(1), 13 - 6 Environmental issues in the planned release of genetically-engineered organisms; Sarma V; This paper examines issues concerned with the environmental release of genetically-engineered micro-organisms . Besides the obvious social and economic benefits from the technology, genetically-engineered micro-organisms can have considerable beneficial effects on environmental concerns, such as the degradation of pollutants and toxic chemical wastes, and less use of hazardous pesticides and chemicals . There may be uncertain negative effects arising from the technology . The hazards of the technology and possible policy approaches are discussed . It is concluded that, overall, the applications arising from this technology are likely to be benign, because its effects on the environment can largely be anticipated by an increase in scientific knowledge gained from field trials and greater experience with releases. Ann Ig, 1989 Jan-Apr, 1(1-2), 81 - 6 {Problems of laundry hygiene}; Signorelli C et al.; As the bacteria found more frequently in hospital infections are the same found in the linen, it is speculated that hospital linens can contribute to the spread of nosocomial patologies . Nevertheless this consideration does not prove that linen constitutes a significant source for transmission of hospital infections, although some studies have strengthened this possibility . The hygienic problems of hospital linens washing have a great importance and in this contest the possibility of contagion throught infected sheets or overalls constitutes only a part of the question . The problem of the spreading of nosocomial infections has to be divided into (1) collection and arlage of the linens; and (2) their disinfection and washing . Collection and carriage need precautions to prevent the hospital staff and the patients from the direct contagion, while during the washing the infected, or potentially infected, linen are to be regarded as a potential risk, which can be avoided through a disinfection before the standard washing cycle . During the washing the processes of cleansing, the dilution and the effect of temperature could be sufficient to obtain the final results . The last consideration is linked to gestational aspects, that's to say the consumptions, the costs, the traditional organizing procedures and the alternative ones, such as the possibility of substituting disposable material to linens or to refer to central laundries placed outside the hospital building and independently operated. Acta Chir Plast, 1989, 31(2), 84 - 91 Recent advances in burn wound management in China; Ding YL et al.; The latest advances in burn wound management in China may be summarized as follows: 1 . Escharectomy, tangential excision and skin grafting is being performed widely in moderate and small deep burns especially in functional sites; immediate flap transfer after debridement for deep third degree burns; tangential excision, escharectomy and large sheet skin grafting along the division lines of the face for deep facial burns . 2 . Micro-skin grafting--a new operative method used in China was introduced to manage extensive deep burns . 3 . Softened freeze-dried glutaraldehyde preserved skin, chlorhexidine-alcohol refrigerated porcine skin, frozen amniotic membrane--all are effective as burn dressings . 4 . The new methods of prolonging the survival time of allo- and xeno-skin grafts are used with ultraviolet rays, medicinal herbs and in vitro treatment with triamcinolone acetonide . 5 . No remarkable rejection occurred after application of composite skin grafts to burn wounds . 6 . The time of culturing epidermal cells has been shortened to 5-13 days . Preliminary successful results were obtained in patients after transplantation of cultured epidermal cell plate or cell emulsion dropped into the holes of allo-skin grafts . No rejection occurred for 50 days up to 16 months after allo-transplantation of epidermal cell plate . 7 . Silver norfloxacin, zinc sulphadiazine and cerium nitrate/silver sulphadiazine had a strong bacteria inhibiting action and promoted wound healing. Int Arch Allergy Appl Immunol, 1989, 90(1), 71 - 7 Production of murine monoclonal antibodies against Francisella tularensis antigens and characterization of antibody-reactive epitopes; Lilliehook B et al.; Eight monoclonal antibodies were produced by immunizing mice with a high molecular-weight antigen from Francisella tularensis . The antigen consists of carbohydrates and proteins and was found to contain several different epitopes . By treating the antigen with periodate or proteinase K, it was found that the majority of antibody-reactive epitopes was mainly dependent on the carbohydrate part of the antigen . On intact F . tularensis bacteria the epitopes seem to be exposed at the bacterial surface . Both main biovars of F . tularensis, tularensis (type A) and palaearctica (type B), exhibited the epitopes. Acta Derm Venereol Suppl (Stockh), 1989, 143, 20 - 30 Effects of azelaic acid on sebaceous gland, sebum excretion rate and keratinization pattern in human skin . An in vivo and in vitro study; Mayer-da-Silva A et al.; The effects of azelaic acid (AZA) on the epidermis of 47 individuals (12 with normal skin, 15 with seborrheic skin and 20 suffering from acne) and on in vitro cultured keratinocytes are reported . Topical application of a 20% AZA cream significantly improved the lesions of acne patients, but failed to induce clinically detectable changes in normal or seborrheic epidermis . Complementary investigations clearly showed that AZA treatment failed to induce specific changes in sebum composition, excretion rate, or in the size of sebaceous glands, but modified epidermal keratinization . Keratohyalin granules and tonofilament bundles were reduced in size and number, mitochondria were swollen and the rough endoplasmic reticulum of malpighian keratinocytes enlarged . The infundibular epidermis of acne individuals showed marked reduction of the horny layer thickness, widening of the horny cell cytoplasm, transitional corneal cells, normalization of filaggrin distribution, and the comedo contained few bacteria and spores . In vitro, AZA exerted marked time- and dose-dependent antiproliferative cytostatic effects on cultured keratinocytes, with a 50% inhibitory dose of 20 mM, decreased some keratinocyte proteins (highly soluble fractions S2, keratohyalin macroaggregate R2, and non-cross-linked fibrous protein S4) and a 95 kD and a 35 kD protein of the cytosolic fraction . Mitochondria were frequently damaged and the rough endoplasmic reticulum enlarged . Our results indicate that AZA is an antikeratinizing agent, displaying antiproliferative cytostatic effects on keratinocytes and modulating the early and terminal phases of epidermal differentiation. Can J Microbiol, 1989 Jan, 35(1), 141 - 7 The use of functional analysis of the ribosome as a tool to determine archaebacterial phylogeny; Amils R et al.; Forty different antibiotics with diverse kingdom and functional specificities were used to measure the functional characteristics of the archaebacterial translation apparatus . The resulting inhibitory curves, which are characteristic of the cell-free system analyzed, were transformed into quantitative values that were used to cluster the different archaebacteria analyzed . This cluster resembles the phylogenetic tree generated by 16S rRNA sequence comparisons . These results strongly suggest that functional analysis of an appropriate evolutionary clock, such as the ribosome, is of intrinsic phylogenetic value . More importantly, they indicate that the study of the nexus between genotypic and phenotypic (functional) information may shed considerable light on the evolution of the protein synthetic machinery. Am J Perinatol, 1989 Jan, 6(1), 41 - 5 Diagnosis of intra-amniotic infection: the acridine orange stain; Romero R et al.; The rapid diagnosis of intra-amniotic infection in patients with premature rupture of the membranes (PROM) and preterm labor is of utmost importance . The Gram's stain examination of amniotic fluid can only detect half of these infections . The acridine orange (AO) stain has been claimed to be superior to the Gram's stain in the detection of bacteria in biologic fluids . Therefore, we undertook the present study to examine the value of AO in the detection of bacteria in amniotic fluid . One hundred and thirty-eight patients with a culture-documented prevalence of intra-amniotic infection of 23.2% were studied . The diagnostic indices of the AO were: sensitivity, 43.8%, specificity, 97.2%, positive predictive value, 82.8%, and negative predictive value, 85.1%; the diagnostic indices for the Gram's stain were: sensitivity, 46.8%, specificity, 98.1%, positive predictive value, 88.2%, and negative predictive value, 85.9% . There was no difference in the diagnostic values of the tests . The agreement between the two techniques was substantial, as indicated by a kappa index of 0.72 (SE = 0.17, p less than 0.001) . The AO offered no significant advantage over the Gram's stain examination of amniotic fluid in the detection of intra-amniotic infection . However, the AO stain was able to identify mycoplasma infections that escaped detection by the Gram's stain . Therefore, the AO is presently the only microscopic technique capable of detecting mycoplasma in amniotic fluid. Biochemistry, 1988 Dec 27, 27(26), 9102 - 8 Reverse gyrase of Sulfolobus: purification to homogeneity and characterization; Nadal M et al.; By using hydrophobic interaction as the first chromatographic stage, we purified to homogeneity reverse gyrase, an ATP-dependent DNA topoisomerase I, isolated from the thermoacidophilic archaebacterium Sulfolobus acidocaldrius . This procedure allowed quick and complete separation of reverse gyrase from nucleases and DNA binding proteins present in Sulfolobus . The final product was revealed, by SDS-PAGE, as a unique band with an apparent molecular mass of 128 kDa, and the amino acid composition was determined . Western blotting experiments with antibodies raised against reverse gyrase indicate that no proteolysis occurred during the purification course . Gel filtration and sedimentation data gave a Stokes radius of 42 A and a sedimentation coefficient of 5.7 S, suggesting a monomeric structure for the native enzyme which was confirmed by electron microscopy . Finally, pure reverse gyrase in a monomeric state was still able to promote positive supercoiling of the DNA. Am J Med, 1988 Dec 23, 85(6A), 59 - 60 Studies of HLA-B27-associated disease; Taurog JD et al.; Several rheumatic diseases were first shown to be associated with human leukocytic antigen (HLA)-B27 in 1973 . Recent developments in understanding this association include the finding that there are at least six variants of HLA-B27 at the molecular level, with no one variant preferentially associated with disease . Detailed studies of the structure of the HLA-B27 molecular family are in progress in several laboratories . Mice expressing HLA-B27 and transmitting it to their offspring (transgenic mice) have been produced and are being studied for their response to bacteria that are known to trigger reactive arthritis in B27+ humans . A particular restriction fragment length polymorphism was recently claimed to be a genetic marker for an additional risk factor in ankylosing spondylitis, but two other laboratories have failed to confirm this finding. Eur J Biochem, 1988 Dec 15, 178(2), 503 - 9 Oxidation-reduction thermodynamics of the acceptor quinone complex in whole-membrane fragments from Chloroflexus aurantiacus; Venturoli G et al.; Oxidation-reduction thermodynamic equilibria involving the quinone-acceptor complex have been examined in whole-membrane fragments from Chloroflexus aurantiacus . The primary quinone acceptor was titrated by monitoring the amount of cytochrome c554 photooxidized by a flash of light as a function of the redox potential . In contrast to previous data obtained in purified plasma membranes, in which the primary quinone acceptor exhibited a midpoint potential equal to -50 mV at pH 8.2, in whole-membrane fragments it titrated at -210 mV (pH 8.0), with a pH dependence of -60 mV/pH up to a pK value of 9.3 . o-Phenanthroline, an inhibitor of electron transfer from the primary to the secondary quinone acceptor, shifted the Em/pH curve of the primary acceptor to higher redox potentials . The midpoint potential of the secondary quinone acceptor and its dependence on pH has been determined by comparing the kinetics of the charge recombination processes within the reaction center complex in the presence and in the absence of o-phenanthroline . It is concluded that both the primary and the secondary quinone acceptors interact with a proton, with pK values of 9.3 and of approximately 10.2 respectively . At physiological pH the electron appears to be stabilized on the secondary with respect to the primary quinone acceptor by approximately 60 meV. Nucleic Acids Res, 1988 Dec 9, 16(23), 11237 - 47 Recombinational resolution in primate cells of two homologous human DNA segments with a gradient of sequence divergence; Gomez-Pedrozo M et al.; Human alpha-thalassemia-2 genotype -alpha 4.2 is the result of meiotic recombination between two 1.3 kb long, homologous DNA segments, X(alpha 2) and X(alpha 1), located in the adult alpha globin locus . The two segments can also undergo intramolecular recombination on extrachromosomal vectors transfected into mitotically dividing primate cells (COS 7) . The existence of a gradient of sequence divergence between X(alpha 2) and X(alpha 1) makes them an interesting system to study the relationship between efficiencies of homologous DNA recombination and the extent of dispersed and localized base mismatches . By partial restriction mapping and DNA sequencing of plasmids recombined in COS 7 cells and rescued from bacteria HB 101, we have determined the distribution of recombinational resolution sites along the two X blocks . Most, if not all, of the homologous recombination events between the two X blocks appear to be single crossing-over without efficient gene correction or repair of base mismatches . The distribution of the sites of recombinational resolution is inversely correlated with that of the gradient of sequence divergence, with only approximately 7% of the X recombinants resolved within the 3' third of the X blocks where two diverged Alu family repeats reside . The Alu sequence within which one of the X recombinants resolved is homologous to a previously characterized alpha thalassemia deletion point. J Mol Biol, 1988 Dec 5, 204(3), 783 - 7 Sulfate-binding protein dislikes protonated oxyacids . A molecular explanation; Jacobson BL et al.; We have determined the effect of pH on the binding affinities of the conjugate bases of four different tetrahedral oxyacids to the sulfate-binding protein . The equilibrium dissociation constants of the binding of sulfate (Kd = 0.12 microM) and selenate (Kd = 5 microM) were found to be pH independent over the range pH 5 to pH 8.1, whereas chromate binding exhibited a pH dependence that is approximately attributable to the pK2 of the chromic acid . Phosphate was bound with an affinity five orders of magnitude weaker than that of sulfate . In light of the highly refined 2 A structure of the complex of the sulfate-binding protein with sulfate, and considering the protonation state and net charge of the various oxyacids, we conclude that the pH dependence of chromate binding and the extremely low affinity of phosphate are attributable mainly to a lack of hydrogen bond acceptors in the binding site . These studies demonstrate that the sulfate-binding site is stringently designed to bind tightly tetrahedral, fully ionized, oxyacid dianions . The presence of a donatable proton on the ligand reduces binding energy by approximately 7 kcal/mol. J Immunol, 1988 Dec 1, 141(11), 3980 - 7 Antiviral immunity induced by recombinant nucleoprotein of influenza A virus . I . Characteristics and cross-reactivity of T cell responses; Tite JP et al.; The gene for the nucleoprotein of the A/NT/60/68 influenza virus was expressed in bacteria and the recombinant protein purified . Lymph node cells from mice immunized with recombinant nucleoprotein proliferated in response to in vitro stimulation with a range of type A influenza viruses . Proliferation was inhibited by mAb to CD4 and class II MHC gene products . IFN-gamma was produced and type-specific CTL were generated in stimulated cultures of immune lymph node cells . These CTL were CD4+ and restricted to class II MHC gene products . Immunization with recombinant nucleoprotein generated Th cells in vivo as measured by the ability to generate an accelerated response to hemagglutinin after challenge with inactivated virus . The results are discussed with reference to a cross-reactive vaccine against influenza. Jpn J Antibiot, 1988 Dec, 41(12), 1841 - 6 {Clinical evaluation of sultamicillin fine granules in lower respiratory tract infection}; Takamoto M et al.; Sultamicillin (SBTPC) fine granules were given to 15 patients with respiratory tract infections at a dose of 375 mg 3 times a day . The results were excellent in 4 patients, good in 8, fair in 1 and poor in 2 with an efficacy rate of 80% which is comparable to that of SBTPC tablet (Unasyn tablet) . One patient complained of difficulty in swallowing the drug probably due to its fine granular nature . No side effects nor abnormal laboratory test values due to this drug were observed in any of the patients enrolled in this study . From the above results, SBTPC fine granules seem to be useful in the treatment of pediatric and elderly patients who sometimes have difficulties in taking tablets. Sci Total Environ, 1988 Dec, 77(2-3), 153 - 61 A study of the sources of external metal contamination of hair; Doi R et al.; Sources of external metal contamination of hair were examined experimentally by exposing hair samples to soil, hot water from a water boiler for domestic use and household dust and fumes in a kitchen . Copper concentration in the hair increased markedly only when the hair was exposed to hot water from the boiler . Iron concentration in the hair increased markedly after exposure to wet soil, and increased slightly after exposure to hot water from the boiler . There was a slight decrease in zinc in the hair after exposure to wet or moist soil, and a significant increase after exposure to hot water from the boiler . When the hair was exposed to household dust and fumes, zinc showed a slight increase but copper and iron showed no change at all . The experiments with soil demonstrated the importance of water in the movement of iron from soil to hair and the role played in this process by biological factors such as soil bacteria. Poult Sci, 1988 Dec, 67(12), 1684 - 93 Respiratory cryptosporidiosis in chickens; Goodwin MA et al.; In order to better characterize spontaneous respiratory cryptosporidiosis in chickens, a retrospective examination of histopathology reports from the Georgia Poultry Laboratories for an 18-mo period (4/1/86 to 9/30/87) was made; 12 cases were found . Collected data were analyzed and certain epidemiologic and histologic features were identified . Eleven of the 12 cases involved broiler type chickens . The ages of chickens with respiratory cryptosporidiosis were evenly distributed between 17 and 52 days of age . The infected birds were always clinically ill . Viruses or bacteria or both often accompanied respiratory Cryptosporidium sp . infections . Histologic lesions (including those of ciliary-adherent bacteria) are described . As the inflammatory response in infected organs became progressively nonpurulent (lymphocytes and plasma cells predominate), numbers of Cryptosporidium diminished . Cytologic preparations were useful for making diagnoses of respiratory cryptosporidiosis in chickens . Identification of epidemiologic features of respiratory cryptosporidiosis, and improved ability to make accurate and prompt diagnoses of Cryptosporidium sp . infection, are vital for a more complete understanding of the impact of this disease on poultry health. Trop Med Parasitol, 1988 Dec, 39 Suppl 4, 466 - 8 The fine structure of male and female Onchocerca gibsoni; Franz M et al.; The fine structures of the midbody regions of male and female Onchocerca gibsoni are described . The epicuticle is wrinkled in the male, and forms long protuberances in the female worm . The lateral chords of the female specimens contain many bacteria and dense bodies . In both sexes the intestinal cells are filled with concentric spherules and the intestinal lumen is extremely reduced . The fine structures of O . gibsoni are very similar to those of O . volvulus. Appl Environ Microbiol, 1988 Dec, 54(12), 3122 - 5 Prevalence of nontuberculous mycobacteria in water supplies of hemodialysis centers; Carson LA et al.; Infection of hemodialysis patients with nontuberculous mycobacteria (NTM) has been associated with water used in reprocessing hemodialyzers . This study was conducted to determine the prevalence of NTM and other bacteria in water samples collected over a 13-week period from 115 randomly selected dialysis centers in the United States . Total viable counts were determined by membrane filter assays; increased recovery of NTM was obtained by dosing a portion of each water sample with 1% formaldehyde (HCHO) before filtering . NTM were widely distributed and occurred with a high frequency in water supplies in dialysis centers . NTM were detected in water from 95 centers (83%), and 50% of all samples examined contained NTM . The results of this study support recommendations to use 4% HCHO or a chemical germicidal equivalent for disinfecting dialyzers that are to be reused. EMBO J, 1988 Dec 1, 7(12), 3625 - 33 Glycine-rich cell wall proteins in bean: gene structure and association of the protein with the vascular system; Keller B et al.; A single genomic clone (14 kb) isolated from bean (Phaseolus vulgaris L.) contains two genes that encode glycine-rich proteins . These genes are present as single copies in the genome, are separated by 2.85 kb and encode transcripts of 1.8 kb and 1.0 kb respectively . The encoded proteins contain 60% glycine and have amino-terminal signal peptides . The 1.8 kb transcript is present in young hypocotyls and in ovary tissue . Excision-wounding transiently induced this transcript in old, but not in young hypocotyl tissue . Antibodies raised against regions of the glycine-rich protein 1.8, expressed as a lacZ fusion protein in bacteria, react with a protein of 53 kd in a protein fraction extracted from cell walls of bean ovaries . Tissue imprints of bean ovaries treated with anti-glycine-rich protein antibodies showed that the glycine-rich protein was distributed in a regular pattern of small, highly localized discrete sites . The immunoreactive regions correspond to the pattern of vascular tissue in the pod . In young hypocotyls, glycine-rich protein is present at four pairs of discrete sites symmetrically arranged on the inner side of the vascular ring . These results suggest a close relationship between glycine-rich proteins and development of the vascular system. Eur J Biochem, 1988 Dec 1, 178(1), 197 - 207 Large-scale preparation and reconstitution of apo-flavoproteins with special reference to butyryl-CoA dehydrogenase from Megasphaera elsdenii . Hydrophobic-interaction chromatography; Van Berkel WJ et al.; A new method is described for the large-scale reversible dissociation of flavoproteins into apoprotein and prosthetic group using hydrophobic-interaction chromatography . Lipoamide dehydrogenase from Azotobacter vinelandii and butyryl-CoA dehydrogenase from Megasphaera elsdenii are selected to demonstrate the usefulness of the method . In contrast to conventional methods, homogeneous preparations of apoproteins in high yields are obtained . The apoproteins show high reconstitutability . The holoenzymes are bound to phenyl-Sepharose CL-4B at neutral pH in the presence of ammonium sulfate . FAD is subsequently removed at pH 3.5-4.0 by addition of high concentrations of KBr . Large amounts of apoenzymes (200-500 mg), showing negligible residual activity, are eluted at neutral pH in the presence of 50% ethylene glycol . The holoenzyme of lipoamide dehydrogenase can be reconstituted while the apoprotein is still bound to the column or the apoenzyme can be isolated in the free state . In both cases the yield and degree of reconstitution of holoenzyme is more than 90% of starting material . Apo-lipoamide-dehydrogenase exists mainly as a monomer in solution and reassociates to the native dimeric structure in the presence of FAD . The apoenzyme is stable for a long period of time when kept in 50% ethylene glycol at -18 degrees C . Steady-state fluorescence-polarization measurements of protein-bound FAD indicate that reconstituted lipoamide dehydrogenase possesses a high stability which is governed by the low dissociation rate constant of the apoenzyme-FAD complex . The holoenzyme of butyryl-CoA dehydrogenase cannot be reconstituted when the apoenzyme is bound to the column . However, stable apoprotein can be isolated in the free state yielding 50-80% of starting material, depending on the immobilization conditions . The coenzyme A ligand present in native holoenzyme is removed during apoprotein preparation . The apoenzyme is relatively stable when kept in 50% ethylene glycol at -18 degrees C . From kinetic and gel filtration experiments it is concluded that the reconstitution reaction of butyryl-CoA dehydrogenase is governed by both the pH-dependent hydrodynamic properties of apoenzyme and the pH-dependent stability of reconstituted enzyme . At pH 7, the apoenzyme is in equilibrium between dimeric and tetrameric forms and reassociates to a native-like tetrameric structure in the presence of FAD . The stability of reconstituted enzyme is strongly influenced by the presence of CoA ligands as shown by fluorescence-polarization measurements . The degree of reconstitution of butyryl-CoA dehydrogenase is more than 80% of the original specific activity under certain conditions.(ABSTRACT TRUNCATED AT 400 WORDS) J Exp Med, 1988 Dec 1, 168(6), 2007 - 21 Tumor necrosis factor/cachectin . Induction of hemorrhagic necrosis in normal tissue requires the fifth component of complement (C5); Rothstein JL et al.; TNF induces hemorrhagic necrosis (HN) when injected into skin exposed to bacterial agents but not when injected into normal skin . In this paper, we present several lines of evidence suggesting that TNF requires the fifth component of complement (C5) to induce HN in skin exposed to bacteria . First, mouse strains that do not have C5 did not develop HN after injection of TNF and bacteria into skin . Second, plasma from C5-sufficient mice could correct the defect in these C5-deficient mice . Third, heating at 56 degrees C for 30 min inactivated the capacity of plasma to reconstitute C5-deficient mice . Fourth, CVF, which is known to inactivate complement, abrogated the capability of C5-sufficient mice to respond . Fifth, depleting plasma of hemolytic activity while generating C5a did not affect the capacity of the activated plasma to reconstitute C5-deficient mice . Finally, only the plasma fraction containing molecules of the size range of C5a reconstituted C5-deficient mice . These findings indicate that C5a and not the membrane attack complex is required for HN . Although we do not know through which mechanism C5a participates in the development of HN, we propose that the described HN response is related to a local defense mechanism in which TNF and C5a lead to the disruption of capillaries in the direct vicinity of bacteria . By this mechanism the rapid spread of bacteria or their products into the circulation is prevented . Such a tissue response is consistent with the known higher susceptibility of C5-deficient mice to bacterial infections and provides a model with which to search for the multiple steps involved in this important local defense mechanism. J Dent Res, 1988 Dec, 67(12), 1505 - 9 Correlation of the hydrolysis of benzoyl-arginine naphthylamide (BANA) by plaque with clinical parameters and subgingival levels of spirochetes in periodontal patients; Schmidt EF et al.; Recent studies have shown that the extent of hydrolysis by plaque of the trypsin substrate, N-benzoyl-DL-arginine-2-naphthylamide (BANA), correlates with the numbers and proportions of spirochetes in subgingival plaque samples, and appears to be an indicator of clinical disease . In this study, BANA hydrolysis by subgingival plaque was evaluated in a blind manner for its ability to reflect both clinical parameters and subgingival levels of bacteria and spirochetes . Subgingival plaque samples were collected from periodontally healthy and diseased sites in 23 untreated periodontal patients and in 13 treated and maintained periodontal patients . In untreated patients, BANA hydrolysis was statistically associated with the total number of spirochetes and bacteria in the plaque sample, but in the treated patients BANA hydrolysis was statistically associated only with the spirochetes . Most BANA-positive reactions in both patient groups were from the sites which were clinically diseased and high in spirochetes . The majority of the negative reactions for BANA hydrolysis in both patient groups was among the sites which were periodontally healthy and low in spirochetes . Specificity and sensitivity of the test were above 80% for disease status in untreated patients . The predictive value of a positive and negative test was above 83% . Slightly lower sensitivity, specificity, and predictive values were found in the treated group . The BANA reaction appears to be an accurate and simple indicator of both clinical disease status and plaque levels of spirochetes in individual tooth sites in untreated and treated periodontal patients. J Dent Res, 1988 Dec, 67(12), 1499 - 504 Methods of disinfection of the water system of dental units by water chlorination; Fiehn NE et al.; The aim of the present study was to develop a simple disinfection method for reducing the content of bacteria in the water system of dental units to an acceptable level . The study was carried out at the Royal Dental College, Copenhagen, on 250 dental units . Samples of the cooling water supplying the ultrasonic scalers and of the water supplying the water glasses were obtained from eight different units representing different parts of the school . Disinfection of the water system was carried out by addition of chlorine to the pipe water near the institution's main water intake . The chlorination of the water was automatically regulated, and the installation was so flexible that the concentration of chlorine and the time and frequency of the chlorination could be varied . Different modes of chlorine dosage were examined . Before chlorination, the bacterial content in the water system of the units was about 10(4)-10(5) cfu/mL . It was found that an intermittent chlorination with 0.5-1 ppm chlorine for 10 minutes every day could reduce the normal bacterial counts in the water system to about a few hundred per mL. J Gen Microbiol, 1988 Dec, 134 ( Pt 12), 3159 - 63 A simple chromatographic procedure for the detection of cyclized archaebacterial glycerol-bisdiphytanyl-glycerol tetraether core lipids; Trincone A et al.; Archaebacterial glycerol-bisdiphytanyl-glycerol tetraether core lipids containing from one to eight cyclopentane rings could be resolved from each other and from the parent uncyclized C40, C40 lipid by TLC . The core lipids of examples from the genera Methanobacterium, Methanobrevibacter, Methanogenium and Methanoplanus did not contain cyclized forms of glycerol-bisdiphytanyl-glycerol tetraethers, whereas the core lipids of Methanosarcina barkeri contained glycerol-bisdiphytanyl-glycerol tetraethers with from one to three cyclopentane rings in each C40 isopranoid chain. Epidemiol Infect, 1988 Dec, 101(3), 669 - 84 Laboratory diagnosis of Mycoplasma pneumoniae infection . 1 . Direct detection of antigen in respiratory exudates by enzyme immunoassay; Kok TW et al.; Direct and indirect antigen capture enzyme immunoassays (Ag-EIA) have been developed for the detection of Mycoplasma pneumoniae in nasopharyngeal aspirates or sputum from respiratory infection . The sensitivity of the two Ag-EIA were similar, but the indirect method using polyclonal rabbit and guinea-pig antisera was more convenient . The Ag-EIA had a detection limit of 10(4-4.5) colony-forming units/ml of sample . It was specific for M . pneumoniae and gave a low level response with M . genitalium . There were no cross-reactions with 10 other species of mycoplasmas . Tests with a wide range of bacteria and chlamydia group antigen, representing agents sometimes found in the respiratory tract, were also negative . At the current level of development, the Ag-EIA detected about 90% of specimens that were also positive for culture; 43% of specimens from culture-negative--seropositive patients gave a positive result . The overall pattern of results indicated that while antigen detection is a quick and effective substitute for the slow culture method, serological examination for specific IgM antibody is also necessary to give a complete diagnostic coverage. J Am Acad Dermatol, 1988 Dec, 19(6), 1067 - 72 Tuberculous ulcer of the skin; Lantos G et al.; A case of tuberculous skin ulcer is reported . The biopsy specimen did not reveal acid-fast bacteria but cultures grew Mycobacterium tuberculosis . A high index of suspicion is needed to diagnose mycobacterial ulcers correctly . The classification of the cutaneous tuberculoses is discussed. Nippon Shishubyo Gakkai Kaishi, 1988 Dec, 30(4), 985 - 95 {High carboxylic acid level in the gingival crevicular fluid (GCF) of the patients with advanced periodontal disease}; Ohwaki K; The carboxylic acids are known to be produced by the bacteria in the dental plaque . The purpose of the present study is to investigate the concentrations of carboxylic acids in the whole saliva and the gingival crevicular fluid (GCF) of periodontal patients . The effects of these acids on human gingival fibroblasts were also examined . Nine males and 9 females patients were divided into two groups (Group A and Group B), according to the differences in probing depth (PD) and alveolar bone loss . Patients of Group A had gingivitis or slight adult periodontitis (PD less than or equal to 4 mm, alveolar bone loss less than or equal to 33%), whereas patients of Group B had moderate to advanced adult periodontitis (PD greater than 4 mm, alveolar bone loss greater than 33%) . Carboxylic acids in the whole saliva and the GCF which were collected from the patients of Group A and Group B were examined using high performance liquid chromatography . The effects of two carboxylic acid salts (butyrate and formate) on the proliferation of human gingival fibroblasts from normal human gingiva were also examined by cell culture . Results obtained were as follows . 1 . The concentrations of four carboxylic acids (acetic, propionic, succinic and butyric acids) in the GCF collected from patients of Group B were significantly higher than that of Group A . On the contrary, all carboxylic acid contents in the whole saliva were not significantly different between the patients of Group A and Group B . 2 . Formate (1 or 3 mM) showed only a small degree of inhibition against the proliferation of human gingival fibroblasts, but butyrate (3 mM) almost completely (95.2%) inhibited the proliferation of human gingival fibroblasts . 3 . The results of the present study may indicate that the carboxylic acids are involved in the initiation and development of the periodontal disease and that they can be one of the indicators in the diagnosis of the periodontal disease. Antibiot Khimioter, 1988 Dec, 33(12), 906 - 9 {Screening of immunomodulators by using the system of mononuclear phagocytes}; Freidlin IS et al.; Immunomodulating effects of various agents can be mediated by the system of mononuclear phagocytes (SMP) . Immunomodulators are able to interfere with the initial events in activation of mononuclear phagocytes (MP) on their membranes which inevitably has its impact on molecular manifestations of the MP activation: the oxidative outburst events . At the initial (tentative) stage of the screening potential immunomodulators it is expedient to use tests providing estimation of the oxidative outburst intensity by the activity of glucoso-6-phosphate dehydrogenase, the key enzyme of the hexose monophosphate shunt and by the level of superoxydanions production judged by reduction of nitroblue tetrasolium (NBTT) . The use of transplantable macrophage-like lines J.744 and P 338D as the targets instead of murine peritoneal MP made it possible to increase reproducibility of the results in screening of immunomodulators . An experimental model of the mouse abdominal cavity clearance within the first hours after intraperitoneal administration of the test bacteria was used for estimating the effect of the potential immunomodulators on the barrier function of the SMP in the host . Experimentally modeled quantitative and functional defects in the SMP served as and adequate test system for study of immunostimulating agents. J Clin Microbiol, 1988 Dec, 26(12), 2598 - 603 Production and characterization of monoclonal antibodies to Mobiluncus species; Fohn MJ et al.; Members of the genus Mobiluncus are anaerobic motile curved rods which are associated with bacterial vaginosis (BV) . Murine monoclonal antibodies (MAbs) to the ATCC type strains of M . curtisii subsp . curtisii, M . curtisii subsp . holmesii, and M . mulieris were produced and characterized by enzyme-linked immunosorbent assay and indirect immunofluorescence assay . Four MAbs were subspecies specific and reacted with M . curtisii subsp . curtisii but not with M . curtisii subsp . holmesii; four were specific for M . mulieris . The remaining antibodies demonstrated some cross-reactivity: three were species specific and reacted with both subspecies of M . curtisii, and one defined an epitope shared by M . curtisii subsp . holmesii and M . mulieris but not by M . curtisii subsp . curtisii . None of the MAbs reacted with a panel of other bacteria commonly present in the vaginas of normal women or women with BV . Examination of the molecular specificities of the antibodies by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblotting revealed four antibodies which were specific for an 82,000-dalton molecule of M . curtisii subsp . curtisii and five antibodies which bound a major band of M . mulieris at 93,000 daltons . Selected MAbs reacted in the indirect immunofluorescence assay with 24 of 25 Mobiluncus spp . clinical isolates from local women with BV and could be used for direct detection of Mobiluncus spp . in vaginal fluid from a patient with BV. J Clin Microbiol, 1988 Dec, 26(12), 2474 - 9 Recognition of mycobacterial antigens by sera from patients with leprosy; Vega-Lopez F et al.; Mycobacterium leprae sonic extracts prepared from armadillo-derived bacteria were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting (immunoblotting) procedures and probed with serum or plasma samples from 20 patients with lepromatous leprosy and 14 healthy endemic controls . Five proteins of 33, 25, 18, 15, and 12 kilodaltons (kDa) were frequently recognized; the 33- and 15-kDa proteins were, respectively, recognized with high intensity by 16 and 13 of the 20 samples from patients with leprosy, whereas only one healthy donor had antibodies that recognized the 15-kDa protein . By the use of M . leprae-specific murine monoclonal antibodies it was demonstrated that the 33-, 25-, and 15-kDa antigens were different from those bound by the available murine monoclonal antibodies . The 18- and 12-kDa proteins detected had molecular masses similar to those detected by the corresponding murine monoclonal antibodies . The serum and plasma samples from patients with leprosy were also used to probe Western blots of a soluble extract of M . tuberculosis . They recognized, among others, antigens with molecular weights similar to those detected in the M . leprae antigenic preparations, although with less intensity and at a lower frequency. Semin Respir Infect, 1988 Dec, 3(4), 291 - 7 Pathology of pleural infections; Harley RA; The pleura is protected by several tissue boundaries of differing degrees of strength . Penetration of the pleural space by mediastinal infection occurs more easily than from the lung, which in turn occurs more easily than through the diaphragm or chest wall . Infectious organisms of all classes, including bacteria, viruses, fungi, and protozoa are capable of causing pleural infection . The basic tissue response in the pleura is similar to that seen in other tissues but is modified by the peculiar anatomy of the pleura, flat serosal surfaces in constant motion against each other . Most pleural infections are initiated in the lung . Some of the most spectacular pathologic lesions are caused by aspergillus where vasculoinvasive fungi cause infected infarcts that result in round visceral pleural lesions . These in turn result in "kissing lesions" of the parietal diaphragmatic surfaces which are of the same size and shape as the visceral pleural lesions, indicating attachment and splinting of the pleural surfaces in these loci . Studies in rabbit demonstrate rapid resolution of experimental empyema with a standardized series of pathologic responses . In uncomplicated empyema, the end result is an extremely thin, almost undetectable scar . Thick fibrotic pleural lesions suggest a complicated course with continuing infection. Transplant Proc, 1988 Dec, 20(6 Suppl 8), 12 - 8 Opportunistic infections in renal allograft recipients; Rubin RH et al.; The risk of opportunistic infection in the renal transplant patient is due to an interaction between two major factors: the epidemiologic exposures (particularly within the hospital environment) and the net state of immunosuppression . The net state of immunosuppression is determined by the nature, dose, and duration of the immunosuppressive therapy being administered; the presence or absence of granulocytopenia and technical factors that could compromise the primary mucocutaneous barriers to infection; such metabolic factors as uremia, hyperglycemia, and the state of nutrition; and, finally, the immunomodulating effects of such viruses as CMV, the hepatitis viruses, and HIV . The major types of opportunistic infection to which the renal transplant patient is susceptible are the following: the viruses of the herpes group and papovaviruses; bacteria such as L monocytogenes, N asteroides, and Legionella; such fungi as Candida, Aspergillus, C neoformans, and the Mucoraceae; and protozoans such as P carinii, S stercoralis, and T gondii. Pediatrics, 1988 Dec, 82(6), 874 - 9 Postoperative necrotizing fasciitis in children; Farrell LD et al.; Necrotizing fasciitis is a rapidly progressive soft tissue infection, involving the skin, subcutaneous tissue, and superficial fascia . It is a rare but life-threatening complication in the postoperative patient . In the last 7 years, we have treated four children in whom necrotizing fasciitis developed after appendectomy for ruptured appendix, bilateral inguinal herniorrhaphy, or gastrostomy closure . These four patients and seven well-described children from the literature with necrotizing fasciitis following surgery form the basis of this review . The ages ranged from six days to 15 years (mean 4.5 years) . There were eight boys and three girls . There were five clean, five clean-contaminated, and one contaminated surgical procedures . No patient had evidence of malignancy or diabetes . Two of our four patients had evidence of failure to thrive . Only one patient had an intraabdominal abscess . In ten, the infection started in the abdominal wall; in one, the infection started in the chest wall . In our four patients, three had neutropenia and fever, four had tachycardia, and two had wound crepitation and radiographic evidence of subcutaneous gas . Cultures of all ten wounds were positive for bacteria; six were positive for more than one organism . Blood culture results were positive in five of five patients who died and in only two of five patients who survived . All survivors had wide surgical debridement and were treated with broad-spectrum antibiotics . The mortality rate was 45% in the whole series.(ABSTRACT TRUNCATED AT 250 WORDS) Infect Immun, 1988 Dec, 56(12), 3189 - 95 Identification of a 69-kilodalton nonfimbrial protein as an agglutinogen of Bordetella pertussis; Brennan MJ et al.; Cells of Bordetella pertussis BP353, a nonfimbriated Eldering serotype 1.3 strain, were used as an immunogen to produce three monoclonal antibodies, BPE3, BPD8, and BPE8, that agglutinated the immunizing cells, as well as certain other nonfimbriated and fimbriated serotype 3-containing B . pertussis strains . The antibodies did not agglutinate serotype 1 or nontypable B . pertussis cells . These monoclonal antibodies specifically detected a 69-kilodalton (kDa) band on Western blots (immunoblots) containing whole B . pertussis cell lysates of Eldering agglutinogen serotypes 1.3, 1.3.6, 1.2.3.4, and 1.2.3.4.6 . This 69-kDa antigen was released from the bacteria by cell incubation for 60 min at 60 degrees C, and it was purified by affinity chromatography with a BPE3-agarose affinity matrix . Purified material was used to produce a polyclonal antiserum that agglutinated all nonfimbriated and fimbriated B . pertussis cells containing serotype 3 agglutinogen . Immunogold electron microscopy and indirect immunofluorescence studies demonstrated that it is an outer membrane constituent but nonfimbrial in appearance . BPE3 did not detect purified fimbriae on Western blots, and antibodies to these fimbriae did not bind to the 69-kDa component . Although B . bronchiseptica and B . parapertussis cells were not agglutinated by the monoclonal antibodies, antigenically similar proteins were detected in extracts of the bacteria . These results identify the 69-kDa protein as a nonfimbrial agglutinogen present on all virulent strains of B . pertussis . The monoclonal antibodies described here should be useful for further studies on the structure and function of this protein. Oncogene, 1988 Dec, 3(6), 665 - 72 Mutational analysis of a phosphotransfer motif essential for v-fps tyrosine kinase activity; Moran MF et al.; The catalytic domains of protein-tyrosine kinases such as the P130gag-fps oncoprotein contain the sequence HRDLAARN, followed thirteen residues C-terminal by DFG (P130gag-fps residues 1041-1048 and 1061-1063) . These residues define a structural motif conserved among eucaryotic protein kinases (-RD----N, DFG) and shared with several procaryotic 3'aminoglycoside phosphotransferases (H-D----N, D-G) . Functional analysis of mutant v-fps proteins employing bacterial and mammalian expression systems indicated that this motif is critical for P130gag-fps kinase activity and oncogenicity . In particular, conservative substitutions of the two invariant aspartates (asp1043, asp1061) with glutamate or asparagine completely eliminated enzymatic activity, suggesting that these residues are essential for catalysis . In contrast, substitution of arg1042 with glutamate decreased but did not eliminate v-fps kinase activity in bacteria . The effects of these and other amino acid substitutions within the phosphotransfer motif and at the nearby autophosphorylation site (tyr1073) of P130gag-fps indicate that these conserved residues are intrinsically essential to the execution or regulation of catalytic activity, and suggest that tight spatial constraints operate within the active centre of the v-fps tyrosine kinase domain. J Biol Buccale, 1988 Dec, 16(4), 219 - 23 Improved SDS-Page method for protein profiles analysis of actinomyces species; Barsotti O et al.; A simple, rapid SDS-Page method for protein profiles analysis of Actinomyces species was developed . Eighteen strains (12 reference strains and 6 fresh isolates) representing four species were used in this study . Eight detergents were tested for protein extraction . Cell extracts were obtained by shaking the bacteria suspended in a solution containing a detergent and glass beads . After electrophoresis, according to the Laemmli's technique, two protein stain procedure were tested (Coomassie blue and silver stain) . Best results were obtained with 0.5% Triton X-100 for 4 h and the silver stain procedure of Oakley . Protein profiles analysis showed that the method is reproducible and distinguishes not only species but sometimes also subspecies. Nucleic Acids Res, 1988 Nov 25, 16(22), 10669 - 79 Identification of psiB genes of plasmids F and R6-5 . Molecular basis for psiB enhanced expression in plasmid R6-5; Dutreix M et al.; PsiB protein of plasmid R6-5 inhibits the induction of the SOS pathway . The F sex factor also carries a psiB gene homologous to that of R6-5 . Yet, it fails to inhibit SOS induction . In order to solve this difference, we characterized the psiB genes of R6-5 and F . We found that (i) the sequences of the two psiB genes share extensive homology the predicted amino acid sequences of the two proteins differing by 5 residues, (ii) the expression of R6-5 psiB is 4 times higher than F psiB gene, (iii) in plasmid R6-5, a Tn10 transposon upstream from the psiB gene enhances psiB expression . Hence, the F sex factor may be unable to prevent SOS induction for two non-exclusive reasons: (i) F PsiB protein, being slightly different from R6-5, may be less active, (ii) the level of synthesis of F PsiB protein may be insufficient to prevent SOS induction. J Immunol Methods, 1988 Nov 25, 115(1), 17 - 29 Phagocytosis of opsonized fluorescent microspheres by human neutrophils . A two-color flow cytometric method for