Bacteria

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 the determination of attachment and ingestion; Ogle JD et al.; A highly reproducible two-color fluorescence cytometric method is described for determining the amount of receptor-mediated and of non-specific phagocytosis by human neutrophils of polystyrene microspheres that have been covalently coated with C3b, iC3b, IgG, mixtures of these, BSA or human F(ab')2 . The method includes a correction for externally bound particles and thus measures net phagocytosis . The method involves the incubation of neutrophils with coated green fluorescent microspheres in buffer alone or with cytochalasin D to inhibit phagocytosis followed by staining the externally bound microspheres with red fluorescent antibodies to the immobilized ligand, and determining the green and red fluorescence in a dual laser fluorescence activated flow cytometer . The red to green fluorescence ratio of the mixtures containing cytochalasin D allows for a correction of the green fluorescence due to externally bound microspheres in the mixtures not containing cytochalasin D . The corrected green intensities thus represent net phagocytosis . The specificity of receptor-mediated phagocytosis was confirmed by inhibition with fluid-phase C3b or iC3b or monoclonal antibodies to the receptors . The method can be applied to the determination of both adherent and suspension phagocytosis and can be used as a general model of the phagocytosis of bacteria by neutrophils.

J Biol Chem, 1988 Nov 25, 263(33), 17251 - 4
Molecular cloning of the beta-subunit of a possible non-F0F1 type ATP synthase from the acidothermophilic archaebacterium, Sulfolobus acidocaldarius; Denda K et al.; The gene which encodes the beta subunit of the novel membrane-associated ATPase has been identified and characterized . The beta subunit, which is most likely the soluble part of the non-F0F1 type H+-ATPase, was obtained from the archaebacterium, Sulfolobus acidocaldarius . In terms of its location, it follows just after the gene for its alpha subunit . It is comprised of 1398 nucleotides, corresponding to a protein of 465 amino acids, and the consensus sequence in the nucleotide binding proteins is poorly conserved . Together with previously described results, the distant homology of the S . acidocaldarius ATPase alpha and beta subunits when compared to those of F0F1-ATPases indicates that this archaebacterial ATPase belongs to an ion-translocating ATPase family uniquely different than F0F1-ATPases even if S . acidocaldarius ATPase and F0F1-ATPases have been derived from a common ancestral ATPase.

Nucleic Acids Res, 1988 Nov 25, 16(22), 10881 - 90
Multiple sequence alignment with hierarchical clustering; Corpet F; An algorithm is presented for the multiple alignment of sequences, either proteins or nucleic acids, that is both accurate and easy to use on microcomputers . The approach is based on the conventional dynamic-programming method of pairwise alignment . Initially, a hierarchical clustering of the sequences is performed using the matrix of the pairwise alignment scores . The closest sequences are aligned creating groups of aligned sequences . Then close groups are aligned until all sequences are aligned in one group . The pairwise alignments included in the multiple alignment form a new matrix that is used to produce a hierarchical clustering . If it is different from the first one, iteration of the process can be performed . The method is illustrated by an example: a global alignment of 39 sequences of cytochrome c.

Biochim Biophys Acta, 1988 Nov 23, 957(2), 301 - 11
Oxalacetate decarboxylase and pyruvate carboxylase activities, and effect of sulfhydryl reagents in malic enzyme from Sulfolobus solfataricus; Guagliardi A et al.; Malic enzyme (S)-malate: NADP+ oxidoreductase (oxaloacetate-decarboxylating, EC 1.1.1.40) purified from the thermoacidophilic archaebacterium Sulfolobus solfataricus, strain MT-4, catalyzed the metal-dependent decarboxylation of oxaloacetate at optimum pH 7.6 at a rate comparable to the decarboxylation of L-malate . The oxaloacetate decarboxylase activity was stimulated about 50% by NADP but only in the presence of MgCl2, and was strongly inhibited by L-malate and NADPH which abolished the NADP activation . In the presence of MnCl2 and in the absence of NADP, the Michaelis constant and Vm for oxaloacetate were 1.7 mM and 2.3 mumol.min-1.mg-1, respectively . When MgCl2 replaced MnCl2, the kinetic parameters for oxaloacetate remained substantially unvaried, whereas the Km and Vm values for L-malate have been found to vary depending on the metal ion . The enzyme carried out the reverse reaction (malate synthesis) at about 70% of the forward reaction, at pH 7.2 and in the presence of relatively high concentrations of bicarbonate and pyruvate . Sulfhydryl residues (three cysteine residues per subunit) have been shown to be essential for the enzymatic activity of the Sulfolobus solfataricus malic enzyme . 5,5'-Dithiobis(2-nitrobenzoic acid), p-hydroxymercuribenzoate and N-ethylmaleimide caused the inactivation of the oxidative decarboxylase activity, but at different rates . The inactivation of the overall activity by p-hydroxymercuribenzoate was partially prevented by NADP singly or in combination with both L-malate and MnCl2, and strongly enhanced by the carboxylic acid substrates; NADP + malate + MnCl2 afforded total protection . The inactivation of the oxaloacetate decarboxylase activity by p-hydroxymercuribenzoate treatment was found to occur at a slower rate than that of the oxidative decarboxylase activity.

J Biol Chem, 1988 Nov 15, 263(32), 16977 - 83
The H circles of Leishmania tarentolae are a unique amplifiable system of oligomeric DNAs associated with drug resistance; White TC et al.; We have induced drug resistance against methotrexate, an inhibitor of dihydrofolate reductase, and CB3717, an inhibitor of thymidylate synthetase in a strain of Leishmania tarentolae . The drug-resistant strains contain extrachromosomal DNA circles of 68 kilobases with a 30-kilobase inverted duplication flanked by 4- and 5 kilobase unique segments . We show that these circles are highly homologous to the drug-induced H circles of L . tropica (1) . All three L . tarentolae strains analyzed contain a chromosomal copy of the H region without duplication, but two of the three strains contain extrachromosomal H circles as well, predominantly present as H circle dimers in one strain and as tetramers in the other . After induction of methotrexate resistance, monomeric circles, presumably derived from the oligomers, become the major type of circle . Our results indicate that the H region represents a genomic region that can be copied at very low frequency to yield circles by a precise, but unusual mechanism under natural conditions in wild-type cells . Although superficially analogous to the episomes of bacteria, the system is without precedent in nature.

Vet Rec, 1988 Nov 5, 123(19), 492 - 4
Haematological findings in budgerigars with megabacterium and Trichomonas infections associated with 'going light'; Henderson GM et al.; Eighteen budgerigars with clinical signs of 'going light' were euthanased and examined post mortem; ingluvitis caused by Trichomonas gallinae infection was present in seven birds, proventriculitis associated with the presence of megabacteria in eight birds and in three birds both conditions were present . Haematological examinations of blood taken from the living birds showed that those with T gallinae infection had normal white cell counts whereas those in which megabacteria were present had significant leucocytosis and heterophilia . Some birds in both groups were anaemic . The findings suggest that infection with megabacteria may be responsible for a proportion of cases of 'going light' in budgerigars and that haematological examination can establish this diagnosis in living birds.

Jpn J Antibiot, 1988 Nov, 41(11), 1751 - 7
{Clinical studies on imipenem/cilastatin sodium in perinatal use}; Chimura T et al.; The clinical efficacy and the safety of imipenem/cilastatin sodium (IPM/CS) in perinatal infections were studied . The results are summarized below . 1 . Clinical efficacy was evaluated in 12 patients with intrauterine infections (endometritis), 3 patients with amniotic fluid infections, 2 patients with puerperal fever and 3 patients with other infections . The drug was administered by intravenous drip infusion at a 0.5g to 1.0g dose as IPM and the total doses during an entire course of treatment were 1.5g to 15g . 2 . Clinical efficacies were excellent in 4 (20%) and good in 16 patients (80%) and the efficacy rate was 100% . Ten patients had not improved upon treatments with other previous antibiotics . 3 . Infective bacteria were eradicated in 4, decreased in 1, and replaced in 6 patients . 4 . No side effects or abnormal clinical laboratory test values were observed in any patient.

J Laryngol Otol, 1988 Nov, 102(11), 1017 - 21
Acute epiglottitis in adults (the Singapore experience); Stanley RE et al.; Acute epiglottitis in adults is a potentially fatal but self-limiting disease of increasing incidence world-wide . Forty-two patients, seen consecutively over a four year period at the ENT Department, Singapore General Hospital were reviewed retrospectively . A strong male predominance with a peak age incidence in the sixth decade was noted . A severe sore throat and dysphagia with disproportionate signs of oropharyngeal inflammation was the main presenting picture . Only three patients had stridor on presentation . Vigilant monitoring of the airway with empirical high-dose intravenous ampicillin, cloxacillin and steroids resulted in a dramatic clinical improvement in most patients and none developed stridor after admission . The yield from throat swabs and blood cultures were low . Two patients developed complications, a Ludwigs angina and an epiglottic abscess . Recurrent epiglottitis was a problem in one patient . There was low morbidity and no mortality on the management regime outlined.

Int J Pept Protein Res, 1988 Nov, 32(5), 406 - 14
Interaction of basic extension peptide fragments of adrenodoxin precursor with phospholipid vesicles; Aoyagi H et al.; Two extension peptide fragments PA1-4 and PA17-32, which correspond to the residues 1-14 and 17-32, respectively, of adrenodoxin precursor, were synthesized by the solution method to find a sequence necessary for the import of the precursor into mitochondria . Biological assay showed that PA1-14 inhibited the import of two mitochondrial enzyme precursors, but PA17-32 showed no inhibition, indicating that the N-terminal sequence has important information for import . CD spectra of the peptides demonstrated that PA1-14 formed alpha-helical structure in Tris-HCl buffer (pH 7.4) containing acidic phospholipid liposomes . Furthermore, PA1-14 induced the moderate leakage of carboxyfluorescein from phospholipid vesicles . The relationship between the structure and function of the peptides is discussed.

Br J Ind Med, 1988 Nov, 45(11), 782 - 9
Recent trends in the prevalence of byssinotic symptoms in the Lancashire textile industry; Cinkotai FF et al.; A respiratory symptoms questionnaire was completed for 4656 volunteers employed in 31 textile factories engaged in spinning or weaving manmade fibre or cotton of various qualities . Sets of airborne dust and bacteria samples were collected in workzones and personal breathing zones in the workrooms where the volunteers were employed . A total of 182 people indicated experiencing byssinotic symptoms, mainly in opening and carding rooms or in spinning and winding rooms where medium to coarse cotton was being processed . This represents a significant decline in the prevalence of byssinotic symptoms over the years, due possibly to lower concentrations of airborne contaminants, especially of bacteria, as cleaner raw materials are being used . According to a multiple, logistic regression model, the prevalence of byssinotic symptoms was found to be statistically significantly related to years worked in the cotton industry, exposure to dust, quality of cotton used, workroom of employment, ethnic origin, and smoking habits . Symptoms of chronic bronchitis were found to be significantly related to smoking habit and to factors connected with occupation, such as exposure to dust, workroom, and the quality of fibre processed.

Surgery, 1988 Nov, 104(5), 846 - 52
Effect of partial burn excision and closure on postburn oxygen consumption; Demling RH et al.; We studied the effect of partial excision and wound closure on the postburn hypermetabolic state . A 25% of total body surface burn was produced in seven sheep . Oxygen consumption (VO2) was significantly increased to 215 +/- 44 ml/min/M2 by day 3 compared with baseline of 125 +/- 21 ml/min/M2 . The calculated increase was the result of the increased cardiac index as the average oxygen (AvO2) difference remained relatively constant . Body temperature was not significantly increased . Plasma and burn lymph thromboxane B2 were significantly increased . On day 7, 60% of the burn was completely excised to fascia and covered with a full-thickness graft from a donor animal . The VO2 decreased to below preburn levels during the period of anesthesia but returned completely to the preexcision hypermetabolic state by 2 hours after anesthesia and remained at this level for the remaining 2-day postexcision period . Quantitative cultures of burn hide at day 7 postburn and of the remaining 10% of total body surface burn at 2 days after excision revealed values less than 10(5) bacteria/gram eschar . No positive blood cultures were evident . We conclude that postburn hypermetabolism, once developed, may be perpetuated by a burn of lesser size . Partial excision, therefore, does not appear to significantly decrease the hypermetabolic state if a substantial inflammatory wound remains . Infection is not necessary to perpetuate the increased VO2.

Proc Natl Acad Sci U S A, 1988 Nov, 85(21), 7927 - 31
Thermotolerance and synthesis of heat shock proteins: these responses are present in Hydra attenuata but absent in Hydra oligactis; Bosch TC et al.; Organisms respond to environmental stress by synthesizing a small number of highly conserved heat shock proteins . In organisms as diverse as bacteria, plants, invertebrates, and vertebrates, synthesis of these proteins is directly correlated with the acquisition of thermotolerance . While studying the freshwater coelenterate hydra, we observed that Hydra oligactis was extremely sensitive to thermal stress . In contrast, the related species Hydra attenuata survives short-term exposure to high temperatures . Furthermore, after incubation at an elevated but nonlethal temperature, H . oligactis did not become thermotolerant . H . attenuata, however, acquired thermotolerance after such a preincubation . In H . attenuata the major heat shock protein was found to be 60 kDa in size . H . oligactis did not synthesize detectable levels of this protein or any new species of proteins in response to stress . Several other species of hydra were found to behave like H . oligactis in response to stress . Thus, these findings provide direct support for the hypothesis that heat shock proteins are required for stress tolerance and that the major heat shock protein in hydra does not have any effects on normal growth or physiology . The findings also indicate that the presence of a heat shock response might be related to the natural environment in which an organism lives.

J Lab Clin Med, 1988 Nov, 112(5), 533 - 43
Use of kinetic methods to evaluate D-xylose malabsorption in patients; Breiter HC et al.; D-Xylose kinetics were studied after administering 25 gm oral and 10 gm intravenous doses to six normal subjects and to 12 patients who were being evaluated for the presence of intestinal malabsorption . D-Xylose absorption was characterized by an absorption rate constant (ka) and a rate constant reflecting nonabsorptive removal of D-xylose from the small bowel (ko) . In normal subjects, mean ka was 0.915 +/- 0.228/hr (+/- SD), and the extent of oral D-xylose absorption averaged 81.0% +/- 11.6% . In seven of the 12 patients, D-xylose absorption was less than 60% complete . In four of them, ka was below the normal limit of 0.367/hr and was consistent with a primary defect in intestinal D-xylose absorption . Two patients with low ka values and the remaining three patients with less than 60% D-xylose absorption had ko values exceeding 0.650/hr, suggesting that excessive nonabsorptive loss contributed to low D-xylose bioavailability . We found that standard tests may identify some patients as having primary defects in D-xylose absorption rate or nonabsorptive loss and propose that breath H2 concentration measurements may also help distinguish between bacterial overgrowth and rapid intestinal transit as causes of excessive nonabsorptive loss.

J Bacteriol, 1988 Nov, 170(11), 5305 - 11
Regulation of carbon flow in Selenomonas ruminantium grown in glucose-limited continuous culture; Melville SB et al.; We have applied a model that permits the estimation of the sensitivity of flux through branch point enzymes (D . C . LaPorte, K . Walsh, and D . E . Koshland, J . Biol . Chem . 259:14068-14075, 1984) in order to analyze the control of flux through the lactate-acetate branch point of Selenomonas ruminantium grown in glucose-limited continuous culture . At this branch point, pyruvate is the substrate of both the NAD-dependent L-(+)-lactate dehydrogenase (LDH) and the pyruvate:ferredoxin oxidoreductase (PFOR) . The LDH was purified, and it exhibited positive cooperativity for the binding of pyruvate . The LDH had an {S}.5 for pyruvate of 0.43 mM, a Hill coefficient of 2.4, and a K' equal to 0.13 mM . The PFOR, assayed in cell extracts, exhibited Michaelis-Menten kinetics for pyruvate, with a Km of 0.49 mM . Carbon flux through the LDH and the PFOR increased 80-fold and 3-fold, respectively, as the dilution rate was increased from 0.07 to 0.52 h-1 in glucose-limited continuous culture . There was nearly a twofold increase, from 6.5 to 11.2 mumol min-1 mg of protein-1 in the specific activity (i.e., maximum velocity) of the LDH at dilution rates of 0.11 and 0.52 h-1, respectively . A flux equation was used to calculate the intracellular concentration of pyruvate; a fourfold increase in pyruvate, from 0.023 to 0.093 mM, was thereby predicted as the dilution rate was increased from 0.07 to 0.52 h-1 . When these calculated values of intracellular pyruvate concentration were inserted into the flux equation, the predicted values of flux through the LDH and the PFOR were found to match closely the flux actually measured in the chemostat-grown cells . Thus, the 80-fold increase in flux through the LDH was due to a twofold increase in the maximum velocity of the LDH and a fourfold increase in the intracellular pyruvate concentration . In addition, the flux through the LDH exhibited ultrasensitivity to changes in both the maximum velocity of the LDH and the intracellular concentration of pyruvate . The flux through the PFOR exhibited ultrasensitivity to changes in the maximum velocity of the LDH and hyperbolic sensitivity to changes in the intracellular concentration of pyruvate.

Immunopharmacology, 1988 Nov-Dec, 16(3), 157 - 65
Stimulatory effects of anti-rheumatic drugs on human neutrophil functions; Oben JA et al.; Auranofin (AF), D-penicillamine (D-pen) and thiola are prescribed as disease-modifying drugs in the treatment of rheumatoid arthritis (RA) . We have shown here that auranofin, 10(-8) to 10(-6) M, D-penicillamine, 10(-6) to 10(-3) M, thiola, 10(-7) to 10(-3) M, and the tripeptide thiol, glutathione, 10(-6) to 10(-3) M, enhanced f-met-leu-phe-induced lysosomal enzyme release and the phagocytic uptake of bacteria by up to 40% . The previously reported inhibitory effects of AF were only observed at concentrations in excess of those likely to be available to effector cells in vivo . The stimulatory effects of thiola and D-pen occurred at concentrations likely to be available to effector cells in vivo and, therefore, may be of greater clinical relevance . There is evidence that the drugs used in this study exert their effects via a thiol moiety and their therapeutic effect is preceded by an elevation of intracellular thiol levels.

Mutat Res, 1988 Nov-Dec, 209(3-4), 155 - 60
The evaluation of genotoxic activities of disinfectants and their metabolites by umu test; Sakagami Y et al.; The genotoxic potential of 6 disinfectants and their 9 metabolites was investigated by umu test . In the tested disinfectants, glutaraldehyde showed positive genotoxicity independent of metabolic activation system and acrinol was positive only in the presence of S9 mixture . Alkylaminoethylglycine, benzalkonium chloride, chlorhexidine digluconate and methylrosaniline chloride were negative in the presence or absence of S9 mixture . In some metabolites of benzalkonium chloride, chlorhexidine digluconate or glutaraldehyde, only pyrogallol showed positive genotoxicity in the absence of S9 mixture and the activity was not affected by the metabolic activation system . Aniline, p-chloroacetoanilide, p-chloroaniline, p-chlorophenol, decabutyldimethylamine, glutaric acid, phenol and pyrocatechol did not induce umu gene expression independently of the presence of S9 mixture . The results in the umu test of these compounds were compared with their findings in the liquid rec-assay and Ames test . The umu test is a more useful and simplified method for the detection of genotoxicity of the compounds with killing effects on tester bacteria.

J Bacteriol, 1988 Nov, 170(11), 5298 - 304
Pathway and sites for energy conservation in the metabolism of glucose by Selenomonas ruminantium; Melville SB et al.; On the basis of enzyme activities detected in extracts of Selenomonas ruminantium HD4 grown in glucose-limited continuous culture, at a slow (0.11 h-1) and a fast (0.52 h-1) dilution rate, a pathway of glucose catabolism to lactate, acetate, succinate, and propionate was constructed . Glucose was catabolized to phosphoenol pyruvate (PEP) via the Emden-Meyerhoff-Parnas pathway . PEP was converted to either pyruvate (via pyruvate kinase) or oxalacetate (via PEP carboxykinase) . Pyruvate was reduced to L-lactate via a NAD-dependent lactate dehydrogenase or oxidatively decarboxylated to acetyl coenzyme A (acetyl-CoA) and CO2 by pyruvate:ferredoxin oxidoreductase . Acetyl-CoA was apparently converted in a single enzymatic step to acetate and CoA, with concomitant formation of 1 molecule of ATP; since acetyl-phosphate was not an intermediate, the enzyme catalyzing this reaction was identified as acetate thiokinase . Oxalacetate was converted to succinate via the activities of malate dehydrogenase, fumarase and a membrane-bound fumarate reductase . Succinate was then excreted or decarboxylated to propionate via a membrane-bound methylmalonyl-CoA decarboxylase . Pyruvate kinase was inhibited by Pi and activated by fructose 1,6-bisphosphate . PEP carboxykinase activity was found to be 0.054 mumol min-1 mg of protein-1 at a dilution rate of 0.11 h-1 but could not be detected in extracts of cells grown at a dilution rate of 0.52 h-1 . Several potential sites for energy conservation exist in S . ruminantium HD4, including pyruvate kinase, acetate thiokinase, PEP carboxykinase, fumarate reductase, and methylmalonyl-CoA decarboxylase . Possession of these five sites for energy conservation may explain the high yields reported here (56 to 78 mg of cells {dry weight} mol of glucose-1) for S . ruminantium HD4 grown in glucose-limited continuous culture.

Przegl Dermatol, 1988 Nov-Dec, 75(6), 435 - 8
{Clinical trial of lysozyme treatment of crural ulcers in humans}; Gasior-Chrzan B; Twenty patients with chronic crural ulcerations refractory to previous treatment were subjected to local treatment with a solution of ovalbumin lysozyme in normal saline (1 mg/ml) . Before this treatment cultures were made from the bottom of the ulcerations determining the type of the infecting micro-organism and its sensitivity to various concentrations of lysozyme . Good (5/20) and very good (15/20) results were obtained . No local or systemic unfavourable effect of lysozyme was noted . The ulcerations were cleared quickly of pus, granulation tissue developed, the inflammatory reaction around the ulcers decreased and pains were no longer felt . The cultured bacteria were insensitive in vitro to lysozyme concentrations applied in vivo . During the treatment with lysozyme the bacterial flora remained unchanged . The beneficial effect of lysozyme on wound healing may be due to its cationic influence on the cell membranes in the epithelium and to pH change in the ulcerations.

J Clin Microbiol, 1988 Nov, 26(11), 2465 - 6
Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity; Hunter PR et al.; An index of discrimination for typing methods is described, based on the probability of two unrelated strains being characterized as the same type . This index may be used to compare typing methods and select the most discriminatory system.

J Comp Pathol, 1988 Nov, 99(4), 421 - 9
A histopathological study of pulmonary infection of mice with Mycobacterium lepraemurium; Ha DK et al.; Intranasal instillation of Mycobacterium lepraemurium (MLM) into mice produced pulmonary infection . MLM multiplied rapidly in the lung tissue during the first few weeks without involvement of other organs . The increase in number, size and confluence of lung granulomas paralleled the multiplication of MLM which could be found both intracellularly and extracellularly . It is postulated that extracellular bacteria may find their way to the bloodstream and thus spread to other visceral organs . Extensive destruction of alveoli and occupation of airspaces by lepra-like cells invariably occurred as the disease progressed.

J Allergy Clin Immunol, 1988 Nov, 82(5 Pt 2), 935 - 40
The role of nasal airway obstruction in sinus disease and facial development; Shapiro GG; Allergic rhinitis in children is often complicated by bacterial sinusitis, which can lead to chronic illness and dysfunction . Sinus disease manifests differently in children than in adults, with cough, rhinorrhea, and middle ear disease being common and pain, headache, and fever being uncommon . Sinusitis may exacerbate asthma, and as many as 70% of children with allergy and chronic rhinitis have abnormal findings on sinus x-ray studies . Nasal cytologic specimens showing large numbers of polymorphonuclear cells with intracellular bacteria are also evidence of sinusitis . Obstruction of the nasal airways by allergic rhinitis or enlarged adenoids can lead to deviations in facial growth, specifically increased facial length . With the removal of the obstruction and a return to nasal breathing, facial length may become more normal . Sinusitis in children is treated with antibiotics, usually for 3 to 4 weeks, to eliminate the infection . Adjunctive therapy with antihistamines, decongestants, cromolyn, and corticosteroids may also be helpful . Topical steroids, such as flunisolide and beclomethasone, can be very useful in pediatric patients . These steroids decrease edema and prevent the release of allergic mediators that may be responsible for an environment favoring the bacterial infection causing sinusitis.

Surgery, 1988 Nov, 104(5), 917 - 23
The gut: a central organ after surgical stress; Wilmore DW et al.; The intestinal tract plays a central role in the protein catabolic response after injury and infection . The mucosa utilizes glutamine and thus spares glucose--presumably sparing this essential fuel source for tissues with an obligate glucose requirement . With inadequate nutritional support or prolonged stress, glutamine levels decrease in both the plasma and the tissue pools, which suggests that glutamine deficiency occurs . This is associated in time with atrophy of the gastrointestinal mucosa . This provision of dietary glutamine results in correction of the abnormally low glutamine concentrations and increased cellularity of the gut mucosa . The derangements in the intestinal mucosa associated with starvation, injury, infection, immunosuppression, chemotherapy, lack of enteral feedings, and other stresses are associated with a breakdown in the barrier function of the gut . Both bacteria and their toxins may enter the host from the intestinal lumen . Through interaction with the reticuloendothelial system, cytokines are produced, which stimulate the pituitary-adrenal axis and thus contribute to the stress response . The elaboration of glucocorticoids facilitates proteolysis, thus increasing glutamine release from skeletal muscle for gut repair . Although this homeostatic mechanism appears to aid mucosal repair and support immunologic responses, severe injury or prolonged glutamine deficits do not adequately support intestinal recovery and allow this cycle to become self-perpetuating (Fig 3) . Adequate enteral feedings initiated early in the course of a disease appear to maintain adequate gut barrier function . In the frequent circumstance when feeding by this route is inadequate or impossible, glutamine-containing parenteral feedings offer an appropriate alternative therapy for bowel and immunologic support . Glutamine-containing parenteral feedings are associated with increased mucosal cellularity and improved survival after gut injury . Specific hormones also stimulate mucosal growth, and it is anticipated that a combination of hormones and specific nutrients will provide optimal support of the gut mucosa in the severely ill patient.

Gastroenterology, 1988 Nov, 95(5), 1351 - 5
Optimization of ascitic fluid culture technique; Runyon BA et al.; The conventional method of ascitic fluid culture detects bacteria in only 42%-65% of patients who have neutrocytic ascites and suspected spontaneous bacterial peritonitis . In this study ascitic fluid was cultured by the conventional method as well as by a new method consisting of bedside inoculation of blood culture bottles with ascites . The conventional cultures grew bacteria in only 13 (43%) of 30 episodes of neutrocytic ascites, whereas the blood culture bottles grew bacteria in 28 (93%); this difference was significant (p less than 0.0001) . The blood culture bottle method also resulted in more rapid detection of bacterial growth . The median concentration of bacteria in infected ascites was one organism per milliliter . Bedside inoculation of blood culture bottles with ascitic fluid is more sensitive than the conventional method in detecting bacterial peritonitis . The insensitivity of the conventional method is probably due to the low concentration of bacteria in infected ascites and the small volume of ascites cultured by this method.

Cell, 1988 Oct 21, 55(2), 361 - 9
Molecular interactions of steroid hormone receptor with its enhancer element: evidence for receptor dimer formation; Tsai SY et al.; A steroid hormone responsive element (GRE/PRE), sufficient to confer glucocorticoid and progesterone inducibility when linked to a reporter gene, was used in band-shift assays to examine its molecular interactions with steroid hormone receptors . Both progesterone and glucocorticoid receptors bound directly and specifically to the GRE/PRE . The purine contact sites for both form A and form B chicken progesterone receptor, as well as those for rat glucocorticoid receptor, are identical . A peptide fragment produced in bacteria that primarily contain the DNA binding domain of the glucocorticoid receptor binds first to the TGTTCT half-site of the GRE/PRE, and a second molecule binds subsequently to the TGTACA (half-site) of the GRE/PRE in a cooperative manner . Utilizing the peptide fragment and the protein A-linked fragment, we demonstrated that the receptor interacts with its cognate enhancer as a dimer.

Lancet, 1988 Oct 15, 2(8616), 872 - 4
Role of pulmonary alveolar macrophage activation in acute lung injury after burns and smoke inhalation; Clark CJ et al.; Bronchoalveolar lavage cells from 42 fire victims and from 18 patients who were smokers attending for diagnostic bronchoscopy (controls) were assessed morphologically and by chemiluminescence . 10 of the victims had inhaled smoke only; 15 had cutaneous burns only; and 17 had combined injury . The combined injury group had significant increases in polymorphonuclear leucocytes and macrophages, especially mature forms, compared with controls . These increases were higher than those expected from the individual injuries . The combined injury group had significantly greater spontaneous chemiluminescence than controls, again greater than that expected by the individual injuries . The chemiluminescence response to stimulation by opsonised bacteria was significantly higher in the combined injury group than in controls, but significantly lower than that in the smoke inhalation only group . The size of the alveolar cellular response to smoke and cutaneous burns suggests that lung damage follows from excess release of inflammatory mediators, exhaustion of the reserve of mature phagocytes and consequent reduced ability to fight bacteria, or both.

Cancer Res, 1988 Oct 15, 48(20), 5624 - 37
Hemopoietic growth factors: a review; Morstyn G et al.; Several hemopoietic growth factors have now been purified, cloned, and produced in bacteria . Granulocyte colony stimulating factor and granulocyte-macrophage colony stimulating factor are already being used in clinical trials . Within 12 months two more hemopoietic growth factors, macrophage colony stimulating factor (also called colony stimulating factor 1) and interleukin 3 (also called multi-colony stimulating factor) will be used for patient treatment . This review discusses the recent advances in our knowledge of the molecular properties and biological specificities of these factors . It is now clear that these molecules are able to modulate selectively the activity of mature blood cells as well as stimulating the production of specific lineages of blood cells . The availability of recombinant hemopoietic growth factors purified from animal or yeast cell conditioned medium or bacteria has facilitated in vivo experiments, as well as the clinical trials . Each of the growth factors has a unique spectrum of biological activities and it appears that the growth factors will enhance the recovery and function of circulating white blood cells after cancer therapy or bone marrow transplantation.

Biochem Biophys Res Commun, 1988 Oct 14, 156(1), 297 - 303
HIV-1 protease specificity of peptide cleavage is sufficient for processing of gag and pol polyproteins; Darke PL et al.; The mature proteins of retroviruses originate as a result of proteolytic cleavages of polyprotein precursors . Retroviruses encode proteases responsible for several of these processing events, making them potential antiviral drug targets . A 99-amino acid HIV-1 protease, produced by chemical synthesis or by expression in bacteria, is shown here to hydrolyze peptides corresponding to all of the known cleavage sites in the HIV-1 gag and pol polyproteins . It does not hydrolyze peptides corresponding to an env cleavage site or a distantly related retroviral gag cleavage site.

J Theor Biol, 1988 Oct 7, 134(3), 319 - 25
A kinetic thermodynamic phenomenological approach to genetic expression of heat-shock proteins; Chela-Flores J et al.; The approach of viewing complex biochemical phenomena as autocatalytic relaxation processes has been introduced previously (Liquori & Tripiciano, 1980; Liquori & Florio, 1985) . In the present work this formalism is extended from its original framework regarding cell growth, to the problem of genetic expression . The case of the heat-shock response in organisms ranging from man to bacteria is discussed . Finally, we give some biochemical examples in which the new approach underlines evident temporal co-operativity.

Biochim Biophys Acta, 1988 Oct 7, 971(3), 233 - 45
Physiological and 15N-NMR analysis of molecular nitrogen fixation by Methanococcus thermolithotrophicus, Methanobacterium bryantii and Methanospirillum hungatei; Belay N et al.; Two mesophilic methanogenic bacteria, Methanobacterium bryantii strain MOH and Methanospirillum hungatei strain GP1 were demonstrated, using several different experimental approaches, to fix dinitrogen . Evidence includes (1) growth with N2 as the sole nitrogen source; (2) incorporation of 15N2 into cellular material (both soluble amino acid pools and insoluble cell protein and other macromolecules) detected by 15N-NMR spectroscopy; (3) acetylene reduction to ethylene by the cells, and inhibition of this reaction by bromoethanesulfonic acid (BES), a methanogen inhibitor . High-resolution 15N-NMR analysis of ethanol extracts of these organisms and cross-polarization magic-angle sample spinning analysis of the solid debris from these extracts are compared to labeled material from Methanococcus thermolithotrophicus, a methanogen previously determined to fix dinitrogen.

J Pediatr, 1988 Oct, 113(4), 641 - 6
C-reactive protein in early detection of bacteremic versus viral infections in immunocompetent and compromised children; Peltola H et al.; The value of quantitatively determined C-reactive protein (CRP), measured from a finger prick sample for rapid detection of septicemia, was examined in 76 blood culture-positive infections in 54 immunocompetent and 18 compromised children; 73 patients with systemic viral infections served as controls . Development of a positive CRP reaction was also studied in 40 cases of acute epiglottitis . Beyond the neonatal age, an increased CRP value (greater than or equal to 20 mg/L) was found in 60 of 64 (94%) children with a positive blood culture for bacteria or fungus . By contrast, CRP remained below this value in 56 of 73 (77%) with viral infections . The immunologic status did not influence the CRP response . However, time had a highly significant (p less than 0.001) effect on CRP; a history of 6 to 12 hours of illness was required before CRP increased above normal . We conclude that CRP is a sensitive and rapidly reacting index in bacteremic infections . However, because other factors than septicemia also increase CRP, we deem a negative CRP value most informative; if two determinations taken several hours apart are less than 20 mg/L, the patient is very unlikely to have invasive bacterial infection.

Xenobiotica, 1988 Oct, 18(10), 1165 - 78
Effects of an NADPH-generating system on primaquine degradation by hamster liver fractions; Abu-El-Haj S et al.; 1 . Primaquine (PQ) often causes severe anaemia in individuals with glucose 6-phosphate dehydrogenase (G6PD) deficient erythrocytes, and metabolites have been implicated as the toxic substance . These studies present data identifying additional metabolites of PQ . 2 . Two metabolites of primaquine (PQ) previously identified in human studies, namely, 6-methoxy-8-aminoquinoline (MAQ) and 8-(3-carboxy-1-methylpropylamino)-6-methoxyquinoline (PQC) were also formed on incubation of PQ with hamster liver fractions for up to 24 h without an NADPH-generating system . 3 . The alcohol (PQAOH) and lactam (PQLT) derivatives of PQ were also formed on incubation with hamster liver fraction used in these studies . 4 . The microsomal metabolism of PQ was decreased in presence of an NADPH-generating system, but not by SKF-525A or glutathione (GSH) indicating that the oxidative reactions were probably not due to the cytochrome P-450 system or free radical mechanisms.

Scott Med J, 1988 Oct, 33(5), 337 - 8
Infective endocarditis as a complication of a permanent indwelling right atrial catheter in a patient with severe aplastic anaemia; Morrison AE et al.; Endocarditis is a recognized complication of both temporary and permanent indwelling right atrial catheters . Endocardial damage by the catheter may result in non-bacterial thrombotic endocarditis . Sterile vegetations composed of platelets and fibrin may become infected, either by direct spread of bacteria along the catheter, or following an episode of bacteraemia . Bacteria in infected vegetations may be protected from phagocytosis by a 'roof' of fibrin . Echocardiography is a valuable non-invasive method of diagnosis and may be used to monitor the resolution of vegetations and valve function . The right atrial catheter produces reflections which must be distinguished from cardiac abnormalities . We report a case of infective endocarditis in a patient with severe aplastic anaemia and a permanent indwelling right atrial catheter which was managed conservatively.

Avian Dis, 1988 Oct-Dec, 32(4), 844 - 8
Small-intestinal cryptosporidiosis in a chicken; Goodwin MA; Severe small-intestinal cryptosporidiosis was diagnosed in a chicken that had diarrhea, was weak, and had failed to thrive . Intestine segments also were parasitized by Eimeria sp . and bacteria . Disseminated foci of malignant lymphocytes were present in several organs . Spontaneously occurring intestinal cryptosporidiosis has not been reported in chickens.

Biochem J, 1988 Oct 1, 255(1), 15 - 21
The primary structures of six human salivary acidic proline-rich proteins (PRP-1, PRP-2, PRP-3, PRP-4, PIF-s and PIF-f); Hay DI et al.; Human glandular salivary secretions contain several acidic proline-rich phosphoproteins (PRPs) . These proteins have important biological functions related to providing a protective environment for the teeth, and appear to possess other activities associated with modulation of adhesion of bacteria to oral surfaces . These functions and activities depend on the primary structures of the PRPs . Previously determined amino acid sequences of two 150-residue molecules, PRP-1 and PRP-2, and two related 106-residue proteins, PRP-3 and PRP-4, indicated that residue 4 was Asn in PRP-1 and PRP-3, and Asp in PRP-2 and PRP-4, and position 50 was Asn in all four proteins . Recent data from cDNA sequence studies and further structural studies, however, showed that the previously proposed sequences cannot be completely correct . The present work has shown that the protein previously designated as PRP-1 actually consisted of two positional isomers, PIF-s, which has Asn and Asp at positions 4 and 50 respectively, and authentic PRP-1, which has the reverse arrangement . The same isomerism is present in the smaller proteins, PIF-f and PRP-3 . Since the isomeric pairs have identical compositions and charges, their presence was not previously detected . Also, by using a more highly purified preparation, it has been found that position 50 in PRP-2 and PRP-4 is Asp, rather than Asn previously reported . These new findings for the six PRPs define their complete primary structures, which are now consistent with those proposed for PRP-1 and PIF-s from cDNA data, and are also consistent with the chromatographic and electrophoretic behaviours of the six PRPs and their derived peptides . These corrected structures are important for understanding the biological functions and activities of these unusual proteins.

Am J Vet Res, 1988 Oct, 49(10), 1688 - 94
Pharmacokinetics and estimated bioavailability of amoxicillin in mares after intravenous, intramuscular, and oral administration; Wilson WD et al.; The pharmacokinetics and estimated bioavailability of amoxicillin were determined after IV, intragastric, and IM administration to healthy mares . After IV administration of sodium amoxicillin (10 mg/kg of body weight), the disposition of the drug was best described by a 2-compartment open model . A rapid distribution phase was followed by a rapid elimination phase, with a mean +/- SD half-life of 39.4 +/- 3.57 minutes . The mean volume of distribution was 325 +/- 68.2 ml/kg, and the mean body clearance was 5.68 +/- 0.80 ml/min.kg . It was concluded that frequent IV administration of sodium amoxicillin would be required to maintain therapeutic plasma concentrations of amoxicillin, and thus, the use of this dosage form should be limited to the initiation of treatment or to intensive care situations . After intragastric administration of amoxicillin trihydrate (20 mg/kg), 5% cherry-flavored suspension, the drug was rapidly, but incompletely, absorbed and rapidly eliminated (mean half-life of the decline phase of the plasma amoxicillin concentration-time curve, 51 minutes) . The mean estimated bioavailability (fractional absorption) of the administered dose was 10.4%, and the mean peak plasma amoxicillin concentration was 2.73 micrograms/ml at 1.5 hours after dosing . In one horse with clinical signs of abdominal discomfort and diarrhea, the absorption of amoxicillin from the gastrointestinal tract was delayed and the fraction absorbed was increased . It was concluded that this oral dosage form could be recommended only for the treatment of infections caused by bacteria that are highly susceptible to amoxicillin, that frequent dosing would be necessary, and that absorption may be inconsistent in horses with gastrointestinal disease.(ABSTRACT TRUNCATED AT 250 WORDS)

Proc Natl Acad Sci U S A, 1988 Oct, 85(19), 7074 - 8
Rhodobacter capsulatus puf operon encodes a regulatory protein (PufQ) for bacteriochlorophyll biosynthesis; Bauer CE et al.; Biosynthesis of the photochemical apparatus by purple nonsulfur photosynthetic bacteria is known to be inhibited by molecular oxygen and high light intensity . Polypeptides that bind bacteriochlorophyll (BChl) to form the light-harvesting I (LH-I) and reaction-center (RC) complexes are encoded by a single transcriptional unit termed the puf operon . In this investigation we demonstrate that the first structural gene in the puf operon (pufQ) of Rhodobacter capsulatus encodes a protein that is required for BChl biosynthesis and that there exists a linear relationship between the amount of pufQ expression and the level of BChl synthesis . Protein sequence similarity exists between PufQ and the region of RC polypeptides that are known to bind BChl and quinone . These observations suggest that pufQ may regulate BChl biosynthesis by a "carrier polypeptide" mechanism as originally proposed by Lascelles.

J Bacteriol, 1988 Oct, 170(10), 4979 - 82
Plasmid DNA from the acetotrophic methanogen Methanosarcina acetivorans; Sowers KR et al.; Nine acetotrophic and three methylotrophic strains of methane-producing bacteria were screened for the presence of plasmid DNA . Plasmids were detected in three marine isolates, including Methanosarcina acetivorans . All three plasmids appeared to be similar based on size and restriction site analyses . The plasmid from M . acetivorans, designated pC2A, was approximately 5.1 kilobase pairs in size and was estimated to be present in a low copy number of six plasmids per genome . Multimers were also observed . A restriction map was constructed . The function of this plasmid is cryptic.

J Bacteriol, 1988 Oct, 170(10), 4652 - 7
Regulation of tetrapyrrole synthesis by light in chemostat cultures of Rhodobacter sphaeroides; Oelze J; Control of bacteriochlorophyll (Bchl), magnesium protoporphyrin monomethyl ester (MgPME), cytochromes, and coproporphyrin by light was studied with chemostat cultures of Rhodobacter sphaeroides growing at a constant dilution rate . By increasing the growth-limiting light energy flux from 10 to 55 W/m2, specific Bchl contents decreased from 19.3 to 7.9 nmol/mg of protein . This was strictly proportional to a decrease in the ratio of B800-850 to B875 light-harvesting complexes . MgPME levels increased from 1.5 to 5.3 nmol/mg of protein, while cytochrome as well as coproporphyrin levels stayed constant at 0.46 and 1.95 nmol/mg of protein, respectively . Since in chemostat cultures steady-state levels of a product represent the rate of synthesis, these results infer only slight control of the rate-limiting step of total tetrapyrrol formation by light . In substrate-limited cultures MgPME was accumulated when growth and Bchl formation approached substrate saturation . This suggests that light controls a second step, i.e., MgPME conversion, whenever too much precursor is available, owing to the low sensitivity of the initial step of control . MgPME was preferentially localized in a subcellular fraction with high contents of B875 complexes . A second fraction exhibiting increased contents of B800-850 complexes lacked significant levels of MgPME . These results are discussed in terms of localization of Bchl synthesis in the membrane system of R . sphaeroides.

Biochem J, 1988 Oct 1, 255(1), 197 - 202
DL-alpha-difluoromethyl{3,4-3H}arginine metabolism in tobacco and mammalian cells . Inhibition of ornithine decarboxylase activity after arginase-mediated hydrolysis of DL-alpha-difluoromethylarginine to DL-alpha-difluoromethylornithine; Slocum RD et al.; DL-alpha-Difluoromethylarginine (DFMA) is an enzyme-activated irreversible inhibitor of arginine decarboxylase (ADC) in vitro . DFMA has also been shown to inhibit ADC activities in a variety of plants and bacteria in vivo . However, we questioned the specificity of this inhibitor for ADC in tobacco ovary tissues, since ornithine decarboxylase (ODC) activity was strongly inhibited as well . We now show that {3,4-3H}DFMA is metabolized to DL-alpha-difluoromethyl{3,4-3H}ornithine {( 3,4-3H}DFMO), the analogous mechanism-based inhibitor of ODC, by tobacco tissues in vivo . Both tobacco and mammalian (mouse, bovine) arginases (EC 3.5.3.1) hydrolyse DFMA to DFMO in vitro, suggesting a role for this enzyme in mediating the indirect inhibition of ODC by DFMA in tobacco . These results suggest that DFMA may have other effects, in addition to the inhibition of ADC, in tissues containing high arginase activities . The recent development of potent agmatine-based ADC inhibitors should permit selective inhibition of ADC, rather than ODC, in such tissues, since agmatine is not a substrate for arginase.

J Parasitol, 1988 Oct, 74(5), 847 - 60
Phylogenetic relationships of the apicomplexan protist Sarcocystis as determined by small subunit ribosomal RNA comparison; Johnson AM et al.; Reverse transcription of total cellular RNA was used to obtain the partial nucleotide sequence of the small subunit ribosomal RNA (srRNA) of Sarcocystis gigantea . The sequence was compared with the homologous sequences of 24 other eukaryotes . Phylogenetic analysis of the semiconserved regions by 4 different tree-building methods using bacteria as an outgroup all concur in showing monophyly of Sarcocystis gigantea and Toxoplasma gondii to the exclusion of all other taxa for which homologous sequences are available.

Ann Cardiol Angeiol (Paris), 1988 Oct, 37(8), 443 - 8
{Cardiac echo-Doppler in the diagnosis and surveillance of the course of acute endocarditis}; Roudaut R et al.; Infectious endocarditis is still currently a problem . Its frequency is not decreasing and new etiologies have developed, which are often responsible for acute endocarditis, resistant to hospital bacteria . The cardiac Doppler has a role in the positive diagnosis of endocarditis, especially on native valves . This examination permits an accurate evaluation of valvular and perivalvular lesions (abscess, mycotic aneurysms) and their repercussions on the ventricle . Studies in this last decade, have emphasized that the presence of vegetations is a factor of gravity . The Doppler permits an "active" monitoring in high risk forms: endocarditis of the aortic orifice, endocarditis with perivalvular abscess, endocarditis occurring on a prosthetic valve.

Toxicology, 1988 Oct, 51(2-3), 281 - 9
Chlorinated drinking water is mutagenic and causes 3-methylcholanthrene type induction of hepatic monooxygenase; Liimatainen A et al.; Acid/neutral fractions of 4 chlorinated drinking water samples were tested for mutagenicity in the Ames' test and injected intraperitoneally to 10- and 20-day-old Wistar rats at doses of 200 and 100 liters of water/kg body weight . Cytochrome P-450 mediated enzyme activities of ethylmorphine-N-demethylase (EMND), 7-ethoxycoumarin-O-deethylase (ECOD), 7-ethoxyresorufin-O-deethylase (EROD) and 7-pentoxyresorufin-O-dealkylase (PEROD) were determined in the 9000 g supernatant fraction of liver homogenate . EROD was introduced by the concentrates . The induction was related to the mutagenic activity . About 4-fold increase in activity was observed with the most mutagenic sample . PEROD was also slightly enhanced . EMND and ECOD activities were not affected by the lower dose, but the higher dose caused inhibition of 30-40% . Although the extracts were not toxic to bacteria, they were unexpectedly toxic to rats . It is concluded that the samples contained 3-methylcholanthrene (3-MC) type inducer(s).

J Exp Med, 1988 Oct 1, 168(4), 1351 - 62
Dissecting human T cell responses against Bordetella species; De Magistris MT et al.; To identify the minimal structures that may be important for the creation of a synthetic and/or recombinant vaccine against whooping cough, human T cell clones were obtained against Bordetella antigens . Cloned peripheral blood T lymphocytes from an immune donor were grown in IL-2 and tested for proliferation in response to inactivated Bordetella species (B . pertussis, B . parapertussis, and B . bronchiseptica) and mutants deficient for the expression of virulence-associated antigens . All the T cell clones obtained were CD4+8- and recognized specifically the Bordetella antigens when presented by autologous B cells . On the basis of the responsiveness to the whole inactivated bacteria, it was possible to cluster the 12 clones obtained into four groups with the following specificity: (1) filamentous hemagglutinin (FHA); (2) B . pertussis-specific antigens; (3) virulence-associated Bordetella-specific antigens; and (4) nonvirulence-associated Bordetella-specific antigens . Using two new B . pertussis deletion mutants, clone 6 (representative of cluster 1) was found to recognize the COOH terminus of FHA . Furthermore, three out of four clones of cluster 3 were specifically stimulated by the soluble 69-kD protein from the outer membrane of B . pertussis . Surprisingly, none of the twelve clones obtained by stimulation in vitro with whole inactivated bacteria recognized pertussis toxin (PT), which is believed to be the most important protein to be included in an acellular vaccine . However, when a new generation of clones was obtained using soluble PT as the in vitro stimulus, it was observed that 11 clones of this group recognized this antigen . Thus, PT does not seem to be the most representative antigen on the whole inactivated bacteria, although T cell memory against PT exists in a donor who had the disease several years ago.

Avian Dis, 1988 Oct-Dec, 32(4), 787 - 92
A comparison of outer membrane proteins and surface characteristics of adhesive and non-adhesive phenotypes of Bordetella avium; Hellwig DH et al.; The outer membrane protein profiles of four adherent and one reduced-adherence mutant phenotype of Bordetella avium were compared; a non-adherent B . avium-like organism isolated from turkeys was also examined . The organisms were grown on brain-heart infusion agar at 35 C for 36 hours . In addition, one of the adherent phenotypes was grown at 18 C and 40 C . The outer membrane proteins were isolated by sonication and detergent extraction with Triton X-100 . Surface characteristics of intact bacteria were examined using negative stain and transmission electron microscopy . The adherent phenotypes had identical protein profiles by electrophoresis . The non-adherent B . avium-like organism lacked at least five of the proteins present on the adherent strains . The non-adherent mutant phenotype had a protein profile similar to that of the adherent organisms, although several proteins were present in much lower concentrations . Fimbriae were found on both adherent and non-adherent organisms . By comparing protein profiles of adherent and non-adherent B . avium we were able to make a preliminary determination of the membrane proteins that lack adhesive properties.

J Bacteriol, 1988 Oct, 170(10), 4681 - 92
In vivo analysis of puf operon expression in Rhodobacter sphaeroides after deletion of a putative intercistronic transcription terminator; DeHoff BS et al.; The intercistronic region of the mRNA derived from the puf operon of Rhodobacter sphaeroides is capable of forming two stable stem-loop structures, the first of which resembles a factor-independent transcription terminator . A puf operon construction lacking the putative transcription terminator was made in vitro and crossed into the chromosome of R . sphaeroides PUFB1 to yield a single chromosomal copy in the terminator-deleted strain . The mutant strain, designated PUF delta 348-420 which was otherwise isogenic with the wild-type strain 2.4.1, showed a normal growth rate at high light intensity compared with the wild type, with the levels of the B875 and reaction center spectral complexes being approximately 7% and 25%, respectively, of those found in the wild type . The deletion mutation correlated with a reduction in the size of the fixed photosynthetic unit from 15:1 in the wild type to 4:1 in the mutant . The level of the B800-850 complex was increased approximately twofold in the mutant strain . However, substantial amounts of the B875 and reaction center polypeptides were not incorporated into spectrally active complexes, suggesting the importance of other factors in the assembly of these complexes . Removal of the intercistronic stem-loops resulted in increased readthrough of the puf operon terminator to regions downstream, as well as altering the stability of the resulting puf operon-specific transcripts . A model is proposed which links ribosome stalling within the open reading frame K leader region of the puf operon transcript with chain termination.

Gene, 1988 Sep 30, 69(2), 337 - 43
Sequence and structural analysis of the alpha- and beta-dinitrogenase subunits of Thiobacillus ferrooxidans; Rawlings DE; The structural genes (nifD and nifK) for the alpha and beta subunits of the molybdenum-iron (MoFe) protein of the Thiobacillus ferrooxidans dinitrogenase have been sequenced . The Mr values deduced from the nucleotide sequences are 54,919 and 57,901 for the alpha and beta subunits, respectively . The amino acid sequences of both subunits were quantitatively compared with the equivalent subunits from other bacteria . Distinct areas of amino acid homology were found between the alpha and beta subunits of T . ferrooxidans.

Nature, 1988 Sep 29, 335(6189), 457 - 8
Phosphonate biosynthesis: isolation of the enzyme responsible for the formation of a carbon-phosphorus bond; Seidel HM et al.; The first isolation of a naturally occurring phosphonate in 1959 led rapidly to the discovery of a variety of metabolites containing a phosphorus-carbon bond . Phosphonates have been found in bacteria, fungi, and higher organisms such as the snail schistosome vector Biomphalaria . The biosynthetic path to the P-C bond has, however, remained undefined . Thus although it was shown twenty years ago that the isotope label from {14C}glucose or from {32P}phosphoenolpyruvate is incorporated into 2-aminoethylphosphonate by the protozoan Tetrahymena pyriformis, the presumed stoichiometric transformation of phosphoenolpyruvate to phosphonopyruvate has never been demonstrated . Low conversions of phosphoenolpyruvate into 2-aminoethylphosphonate and the trapping of phosphonopyruvate from phosphoenolpyruvate have been reported, but these reactions have not proved reproducible, and the existence of the critical enzyme, phosphoenolpyruvate phosphonomutase, has remained notional . We now report experiments that resolve this enigma, and describe the isolation and characterization of the pure mutase from T . pyriformis.

Biochemistry, 1988 Sep 20, 27(19), 7531 - 7
Characterization of the ATP synthase of Propionigenium modestum as a primary sodium pump; Laubinger W et al.; The ATP synthase (F1F0) of Propionigenium modestum has been purified to a specific ATPase activity of 5.5 units/mg of protein, which is about 6 times higher than that of the bacterial membranes . Analysis by SDS gel electrophoresis indicated that in addition to the five subunits of the F1 ATPase, subunits of Mr 26,000 (a), 23,000 (b), and 7500 (c) have been purified . The ATPase activity of F1F0 was specifically activated about 10-fold by Na+ions . The enzyme was strongly inhibited by dicyclohexylcarbodiimide, venturicidin, tributyltin chloride, and azide . After incubation with {14C}dicyclohexylcarbodiimide, about 3-4 mol of the inhibitor was bound per 500,000 g of the enzyme . The radioactive label was specifically bound to submit c . These subunits form stable aggregates which resist dissociation by SDS at 100 degrees C . The monomer is formed upon heating with SDS to 121 degrees C or by extraction of the membranes with chloroform/methanol . The ATP synthase was incorporated into liposomes by a freeze-thaw-sonication procedure . The reconstituted proteoliposomes catalyzed the transport of Na+ions upon ATP hydrolysis . The transport was completely abolished by dicyclohexylcarbodiimide . Whereas monensin prevented the accumulation of Na+ions, the uptake rate was stimulated 4-5-fold in the presence of valinomycin or carbonyl cyanide m=chlorophenylhydrazone . These results indicate an electrogenic Na+ transport and also that it is a primary event and not accomplished by a H+-translocating ATP synthase in combination with a Na+/H+ antiporter.

J Biol Chem, 1988 Sep 15, 263(26), 13310 - 6
Bordetella pertussis adenylate cyclase . Identification of multiple forms of the enzyme by antibodies; Rogel A et al.; Two forms of Bordetella pertussis adenylate cyclase of 200 and 47 kDa have been purified from dialyzed urea extract of the bacteria to specific activities of 466 and 1685 mumol.min-1.mg-1, respectively . Both forms are activated 50-200-fold by calmodulin . The half-maximum concentration required for the activation of the 200-kDa catalyst is 5.4.10(-9) M, whereas the one required for activation of the 47-kDa catalyst is 1.8.10(-10) M . Polyclonal antibodies raised against the 47-kDa catalyst specifically recognize both forms of the enzyme in purified state as well as in bacterial extracts on immunoblots . The antibody inhibits at similar titer adenylate cyclase activity of the purified forms as well as the activity present in dialyzed urea extract of the bacteria . It also prevents the penetration of the invasive B . pertussis adenylate cyclase into human lymphocytes . The inhibition induced by the antisera is specific to B . pertussis enzyme, since both calmodulin-dependent brain and sperm adenylate cyclase are not affected by the antibody.

J Immunol Methods, 1988 Sep 13, 112(2), 183 - 90
Purification of human monocytes/macrophages by adherence to cytodex microcarriers; Kumaratilake LM et al.; Adherence to cytodex microcarries was used as a means of purifying and culturing human monocytes . Mononuclear leukocytes obtained from centrifugation of blood on Hypaque-Ficoll medium could be enriched for monocytes/macrophages by adherence to cytodex microcarriers . Using this system it was possible to obtain monocytes/macrophages in high yields and viability, and good purity . Detachment of the monocytes from the microcarriers was simply achieved by a 1-2 min agitation of the contents . Cells which had been cultured up to 30 days on the microcarriers could be similarly detached . Functional studies on the detached monocytes macrophages showed the cells to respond well to stimuli such as zymosan and fMLP, and were capable of killing opsonized bacteria.

FEBS Lett, 1988 Sep 12, 237(1-2), 91 - 7
L-tyrosine is the precursor of PQQ biosynthesis in Hyphomicrobium X; van Kleef MA et al.; A method was developed to study amino acids as possible precursors of PQQ biosynthesis . Cultures of Hyphomicrobium X, growing on {13C}methanol, were supplemented with unlabelled amino acids . Uptake and participation in metabolism were determined via gas chromatography/mass spectrometry of derivatized amino acids, obtained from hydrolyzed cellular protein, by measuring their 12C content . Several amino acids appeared to be incorporated into the protein to a significant extent, without degradation or conversion . Among these were the aromatic amino acids, L-tyrosine and L-phenylalanine . Using the same replacement approach, their incorporation into PQQ was determined by 1H- and 13C-NMR spectroscopy of purified PQQ obtained from the culture medium . It appeared that the complete carbon skeleton of tyrosine was present, forming the o-quinone and pyrrole-2-carboxylic acid moieties in PQQ, while phenylalanine was not incorporated at all . Starting with L-tyrosine, possible biosynthetic routes to PQQ are discussed.

FEBS Lett, 1988 Sep 12, 237(1-2), 141 - 4
A stretch-activated anion channel in tobacco protoplasts; Falke LC et al.; Stretch-activated ion channels have been described in animal cells, where they might serve as mechanoreceptors, baroreceptors or osmoreceptors, as well as in yeast and bacteria, where osmoregulatory functions have been suggested . Here we report a large conductance, stretch-activated, anion-selective channel in protoplasts of a higher plant, tobacco, and discuss its possible role in osmoregulation.

Nature, 1988 Sep 8, 335(6186), 176 - 8
Vaccinia virus encodes a secretory polypeptide structurally related to complement control proteins; Kotwal GJ et al.; Several polypeptides are secreted into the medium of cells infected with vaccinia virus, a cytoplasmic DNA virus belonging to the poxvirus family . One of these, a polypeptide of relative molecular mass 19,000 is structurally related to epidermal growth factor and binds to epidermal growth factor receptor stimulating proliferation of uninfected cells in vitro and in vivo . Here, we show that a second, and much more abundant secretory polypeptide, is also encoded by vaccinia virus and is structurally related to the superfamily of complement control proteins . Members of this family can block complement-mediated induction of the inflammatory response, and engulfment, killing and lysis of bacteria and viruses.

Gene, 1988 Sep 7, 68(2), 323 - 33
Use of pulsed-field-gradient gel electrophoresis to construct a physical map of the Caulobacter crescentus genome; Ely B et al.; The restriction enzyme DraI cleaves the Caulobacter crescentus genome into at least 35 fragments which have been resolved in agarose gels using pulsed-field-gradient gel electrophoresis (PFGE) . When digests were performed using DNA from strains containing Tn5 insertion mutations, altered band migrations were observed . Using PFGE with the appropriate pulse times, size differences as small as 2% could be resolved in large fragments . Using this approach, we have constructed a partial physical map of the genome which correlates well with the C . crescentus genetic map and have shown the size of the genome to be approx . 3800 kb . Using hybridization with cloned genes, we have determined the map locations of five previously unmapped genes . In addition, we have shown that PFGE can be used to rapidly determine the map locations of new insertion mutations or the sizes of deletion mutations.

J Thorac Cardiovasc Surg, 1988 Sep, 96(3), 436 - 9
The role of early limited thoracotomy in the treatment of empyema; Van Way C 3rd et al.; On the basis of clinical experience with 80 patients at Denver General Hospital from 1979 through 1984, we devised a three-part classification of empyema . Class I empyema (n = 12) is pleural effusion with pH less than 7.2 and with no bacteria . Patients with this type of empyema were treated with short-duration chest tubes . Further treatment was required in two of 12 . There were no deaths . Class II (n = 28) is classic uniloculate empyema . Patients with this category of empyema were treated with chest tubes, with two deaths . Class III (n = 40) is complicated empyema, with multiple loculations . Tube thoracostomy failed more often than not; decortication was required in 10 of 18 patients treated with prolonged tube drainage . Limited thoracotomy for drainage and placement of tubes was done in 22 patients . Five required extension of the thoracotomy and decortication . All 22 had resolution of the empyema with no additional procedures . Limited thoracotomy immediately or during the first week of treatment is recommended for all multiloculated or complex empyemas.

J Bacteriol, 1988 Sep, 170(9), 3915 - 23
Phenotypic switching of Thiobacillus ferrooxidans; Schrader JA et al.; Two solid medium formulations, designated 100:10 and 10:10, were developed for the growth of Thiobacillus ferrooxidans . The new media contain a mixture of both ferrous iron and thiosulfate as available energy sources, permitting the detection of colony morphology variants that arise spontaneously in a wild-type population . Several morphological and physiological characteristics of a class of T . ferrooxidans variants, termed LSC for large spreading colony, are described . LSC variants lack the ability to oxidize iron but retain the capacity to utilize thiosulfate or tetrathionate as energy sources . An LSC colony spreads on the surface of solid 100:10 medium as a monolayer of cells in a fashion resembling that of certain swarming or gliding bacteria . The LSC variant reverts to a parental wild type at frequencies that vary in different independently arising isolates . The identity of the LSC variant as a derivative of the parental wild-type T . ferrooxidans was established by Southern blot hybridization.

J Bacteriol, 1988 Sep, 170(9), 3882 - 90
Quantitative assay for binding of Bradyrhizobium japonicum to cultured soybean cells; Ho SC et al.; Incubation of Bradyrhizobium japonicum with the cultured soybean cell line SB-1 resulted in the adhesion of the bacteria to the plant cells . An antiserum was raised against B . japonicum, and the 125I-labeled immunoglobulin fraction was used to quantitate the number of bacteria bound to the soybean cells . The measurement of 125I-labeled antibody binding correlated well with parallel assays by microscopic observation . Using this quantitation, we have optimized the parameters of the assay in terms of time course, ratio of B . japonicum to SB-1 cells, and pH . We then explored the effects of saccharides, NaCl, EDTA, and culture age of the bacteria and SB-1 cells on B . japonicum binding under these optimal assay conditions . The results showed good correlation between conditions that govern B . japonicum binding to SB-1 cells in culture and those that regulate B . japonicum-induced nodulation in legume roots . Together, they suggest that this binding event may be important in controlling host specificity.

Dev Biol, 1988 Sep, 129(1), 114 - 23
Accumulation and decay of DG42 gene products follow a gradient pattern during Xenopus embryogenesis; Rosa F et al.; The DG42 gene is expressed during a short window during embryogenesis of Xenopus laevis . The mRNA for this gene can be first detected just after midblastula, peaks at late gastrula, and decays by the end of neurulation . The sequence of the DG42 cDNA and genomic DNA predicts a 70,000-Da protein that is not related to any other known protein . Antibodies prepared against portions of the DG42 open reading frame that had been expressed in bacteria detected a 70,000-Da protein in the embryo with a temporal course of appearance and decay that follows that of the RNA by several hours . Localization of the mRNA in dissected embryos and immunohistochemical detection of the protein showed that DG42 expression moves as a wave or gradient through the embryo . The RNA is first detected in the animal region of the blastula, and by early gastrula is found everywhere except in the outer layer of the dorsal blastopore lip . By midgastrula DG42 protein is present in the inner ectodermal layer and the endoderm; it disappears from dorsal ectoderm as the neural plate is induced and later decays in a dorsoventral direction . The last remnants of DG42 protein are seen in ventral regions of the gut at the tailbud stage.

Cell Immunol, 1988 Sep, 115(2), 221 - 7
Prevention by a platelet-derived factor (platelet factor 4) of induction of low dose tolerance to pneumococcal polysaccharides; Yin JZ et al.; It was previously shown that human or mouse serum, and platelet factor 4 (PF4) prepared from human platelet releasate, counteracts nonspecific immunosuppression induced in mice by injection of concanavalin A or syngeneic gamma-irradiated lymphoma cells . The present studies show that PF4 prepared from normal mouse or human serum by absorption to heparin-agarose and elution between 0.5 and 1.5 M NaCl is also active in this respect . The ability of PF4 to counteract antigen-specific suppression of the antibody response to pneumococcal polysaccharide (pps) was now studied . PF4 derived from human or mouse serum as well as recombinant PF4 interferes with induction of antigen-specific low dose tolerance when they are injected at the same time as a low dose (0.2 microgram) of type 14 pps 3 days before an optimal immunizing dose (25 micrograms) . Furthermore, injection of platelet releasate at the time of an optimal primary immunizing dose of pps type 14 enhances the secondary response to killed bacteria injected 2 weeks later, but not the primary response itself . Both effects are interpreted as due to interference with antigen-specific suppressor cell induction during primary immunization . Injection of PF4 is much less effective in reversing low dose tolerance to an optimal immunizing dose (0.1 microgram) of type 3 pps induced by injection of 0.005 microgram of this antigen . Differences in the mechanism of tolerance induction for the two pps types that might be responsible for this are discussed.

Scand J Gastroenterol, 1988 Sep, 23(7), 808 - 12
Experimentally induced ileal ulcers in rats . A model to study non-specific inflammatory bowel disease; Wickbom G et al.; A surgical technique was used to establish chronic intestinal ulcers in Sprague-Dawley rats . A 2-cm-long segment of the distal ileum was excised and left attached on the mesentery . The ileum was reanastomosed . The excluded ileal segment formed a 'cyst' of various sizes . Initially, the anastomoses healed well, but after 6-8 weeks para-anastomotic ulcers developed in more than 50% of the rats . Histopathology showed a chronic inflammatory reaction with a predominance of mononuclear cells and increased numbers of eosinophilic granulocytes . The surface of the ulcers was covered with bacteria . Penetrating ulcers with fistula formation occurred . It is concluded that this experimental model may be useful for time sequential studies of the development of chronic and ulcerative ileitis . It may also be used to study the effect of medical and surgical regimens for the treatment of non-specific chronic inflammatory bowel disease.

Food Chem Toxicol, 1988 Sep, 26(9), 797 - 801
The metabolism and deamination of {14C}sulphamethazine in a germ-free pig: the influence of nitrate and nitrite; Struble CB et al.; Twenty-four hours after feeding nitrite in combination with {14C}sulphamethazine to a germ-free and a conventional control pig the level of {14C}desaminosulphamethazine in tissues from both pigs was high, accounting for 11 to 30% of the total tissue 14C . When another germ-free pig was fed {14C}sulphamethazine in combination with nitrate, a trace amount of {14C}desaminosulphamethazine was found by gas chromatography-mass spectroscopy in the skeletal muscle but not in other tissues . When a control pig was fed {14C}sulphamethazine and nitrate, {14C}desaminosulphamethazine was found in all tissues examined . The results from this study show that feeding pigs nitrite together with sulphamethazine increases the amount of desaminosulphamethazine in the tissues . Most of the desaminosulphamethazine found in the tissues of the control pig fed nitrate was presumably the secondary effect of bacterial reduction of nitrate to nitrite.

J Biol Buccale, 1988 Sep, 16(3), 157 - 68
{Evaluation of the Caridex system and its pulp biocompatibility}; Waltmann E et al.; A study of the pulpal biocompatibility of the Caridex system was carried out on human premolars to be extracted bilaterally for orthodontic reasons, in 3 volunteers aged 10 to 13 years . A class V cavity was prepared bilaterally on the 2 premolars to be extracted: one of the cavities was washed with Caridex . The other was used as a control . The extractions were made immediately, 4 and 7 weeks after these operative procedures . Histological examinations showed that the Caridex system was biocompatible for the human dental pulp . A comparative study of carious cavities was conducted using light microscopy, Gram stain and transmission electron microscopy on the quality of the cleaning obtained after conventional cavity preparation and use of the Caridex system . After Caridex treatment, the dentinal surface was free of demineralized zones and no smear layer was present . The occasional presence of bacteria was noticed after conventional use of a bur as well as after Caridex . More limited clinical indications of the Caridex system are proposed.

Am J Sports Med, 1988 Sep-Oct, 16(5), 539 - 40
Ethyl chloride: an ineffective bacteriostatic or cidal agent for arthrocentesis; James WS 3rd et al.; The local and general anesthetic properties of ethyl chloride have been well delineated . The purpose of this study was to determine if ethyl chloride, when used as a local anesthetic, had any bacteriostatic or cidal effects on the normal array of skin flora . Fifteen different aerobic bacteria or fungi were collected from pure cultures . A total of 30 specimens were sprayed; 2 specimens of each of the 15 bacteria were sprayed with ethyl chloride for both 5 and 10 seconds . These specimens then were restreaked once the ethyl chloride had evaporated, and were incubated for 24 hours . At 24 hours the cultures were checked and found to have no significant growth inhibitions . It is thus concluded that ethyl chloride alone does not have any bacteriostatic or cidal effects on these commonly found skin flora.

Equine Vet J, 1988 Sep, 20(5), 368 - 70
Effect of ovarian steroids on migration of uterine lumenal neutrophils and on chemokinetic factors in uterine secretions from mares; Watson ED; Incubation of blood neutrophils with uterine flushings collected from ovariectomised mares treated with oestradiol, stimulated migration under agarose, whereas flushings from mares treated with progesterone or oily vehicle, inhibited migration . After intra-uterine infusion of bacteria, however, flushings from oestradiol-treated and vehicle-treated mares inhibited migration, whereas progesterone treatment stimulated migration . Migration of uterine-derived neutrophils under agarose was less than that of blood neutrophils and was not influenced by treatment with ovarian steroids . Uterine susceptibility to infection in progesterone-treated mares was not, therefore, correlated with the migratory activities of uterine neutrophils or the chemokinetic properties of uterine secretions in vitro.

Anaesthesia, 1988 Sep, 43(9), 760 - 1
Minitracheotomy in children . The management of sputum retention in older children by minitracheotomy; Allen PW et al.; The successful management of sputum retention by minitracheotomy in two 12-year-old patients is reported . Some advantages and potential problems are discussed . This is the first time that minitracheotomy has been reported in this age group.

Acta Radiol, 1988 Sep-Oct, 29(5), 565 - 70
Chronic recurrent multifocal osteomyelitis in children . A roentgenologic and scintigraphic investigation; Mortensson W et al.; Thirty cases of chronic recurrent multifocal osteomyelitis have been reported in the literature . A radiologic evaluation of thirty-one additional cases, 25 of whom also underwent bone scintigraphy, is presented . Bone biopsy specimens were obtained in 16 patients . Most lesions were located in the metaphyses of the long bones adjacent to the physis and had a characteristic, probably pathognomonic appearance . Extension into the epiphyses was rare . Lesions in the vertebral bodies, clavicle and pelvis had possibly a less specific radiographic appearance . Lesions in short tubular bones were non-specific . Bone scintigraphy had a practical value in evaluating the global distribution of lesions including asymptomatic lesions and lesions in the spine or pelvis, the latter being somewhat hard to detect with conventional radiography . All biopsies showed acute, subacute or chronic unspecific osteomyelitis, sometimes mixed in the same lesion . Staining for bacteria and fungi was negative.

J Gen Microbiol, 1988 Sep, 134 ( Pt 9), 2605 - 14
Proteinase activity in rumen ciliate protozoa; Lockwood BC et al.; Azocasein-degrading proteinase activity was detected in all rumen ciliate protozoa that were examined from four entodiniomorphid and two holotrich genera . All of the activities were optimal in the range pH 4.0-5.0 and were inhibited by cysteine proteinase inhibitors, notably leupeptin . The inhibition profiles and extent of inhibition observed with the different groups of inhibitors were organism-specific . Gelatin-SDS-polyacrylamide gel electrophoresis of protozoal lysates revealed multiple forms of the proteinases in the species examined . The number of enzymes detected, their molecular masses, the level of activity and inhibitor susceptibility was genus-dependent . The proteinase profiles of the two holotrich species differed and inter-species differences were also apparent among species of the genus Entodinium . The characteristics and molecular size distribution of rumen bacterial proteinases were different to the protozoal proteinases . Low levels of proteinase activity, of apparently bacterial origin, were detected by gelatin-SDS-PAGE analysis of cell-free rumen liquor.

Infect Dis Clin North Am, 1988 Sep, 2(3), 705 - 18
Diarrhea in AIDS; Lopez AP et al.; The spectrum of illness associated with AIDS has been enlarging since its initial description in 1981, and the gastrointaestinal tract continues to be one of the major targets of this devastating disease . The many causative agents are discussed in this article, including bacteria, viruses, fungi, protozoa, helminths, and various noninfectious diseases.

Genetics, 1988 Sep, 120(1), 23 - 35
Reciprocality of recombination events that rearrange the chromosome; Mahan MJ et al.; We describe a genetic system for studying the reciprocality of chromosomal recombination; all substrates and recombination functions involved are provided exclusively by the bacterial chromosome . The genetic system allows the recovery of both recombinant products from a single recombination event . The system was used to demonstrate the full reciprocality of three different types of recombination events: (1) intrachromosomal recombination between direct repeats, causing deletions; (2) intrachromosomal recombination between inverse homologies, causing inversion of a segment of the bacterial chromosome; and (3) circle to circle recombination (in the absence of any plasmid or phage functions) . Results suggest that intrachromosomal recombination in bacteria is frequently fully reciprocal.

Acta Paediatr Scand, 1988 Sep, 77(5), 747 - 51
Effect of prophylactic acetaminophen administration on reaction to DTP vaccination; Uhari M et al.; The effectiveness of acetaminophen in preventing post-vaccination fever was studied in a double-blind randomized manner . Healthy five-month-old infants vaccinated with diphtheria-tetanus-pertussis (DTP) or DTP-inactivated polio vaccine were randomly allocated to receive either placebo (n = 130) or 75 mg of acetaminophen (n = 133) four hours after the vaccination . Rectal temperatures of the infants were measured in the post-vaccination evening and next morning by the parents . The mean values of rectal temperatures were equal in both groups, i.e . 37.6 degrees C, both in the evening and in the morning . No significant difference was found in the occurrence of other minor adverse reactions . Antibody titres to diphtheria and tetanus toxoids and pertussis bacteria of the placebo (n = 25) and acetaminophen (n = 34) groups did not differ significantly from each other . It is concluded that acetaminophen in a single dose schedule is ineffective in decreasing post-vaccination fever and other symptoms.

Arch Latinoam Nutr, 1988 Sep, 38(3), 519 - 42
{Carbohydrates and fiber}; Lajolo FM et al.; Dietary carbohydrates comprise two fractions that may be classified as digestible, and which are useful as energy sources (simple and complex carbohydrates) and fiber, which is presumed to be of no use to the human body . There are insufficient epidemiologic data on the metabolic effects of simple carbohydrates and it is not advisable to make quantitative recommendations of intake . It is questionable to recommend in developing countries that a fixed proportion of dietary energy be derived from simple sugars, due to the high prevalence of deficient energy intake, cultural habits, and regional differences in food intake and physical activity . In relation to recommendations of complex carbohydrates, it should be considered that their absorption is influenced by many factors inherent to the individual and to the foods . Fiber is defined as a series of different substances derived from tissue structures, cellular residues and undigested chemical substances that may be partially utilized after intestinal bacteria have acted on them . There is not a clear definition of the chemical composition of fiber, but it consists mainly of polysaccharides (such as cellulose, hemicellulose and pectins), lignin and end products of the interactions of various food components . The effects of fiber, such as control of food intake, regulation of gastrointestinal transit, post-prandial blood concentrations of cholesterol, glucose and insulin, flatulence and alterations in nutrient bioavailability are due to various physical properties inherent to its chemical components . Impairment of nutrient absorption may be harmful, mainly among populations whose food intake is lower than their energy needs, and with a high fiber content . This may be particularly important in pregnant women, growing children and the elderly, and should be considered when making nutrient recommendations . A precise knowledge of fiber is also important to calculate the real energy value of foods, mainly for two reasons: 1) the proportion of "crude fiber" (as measured by acid and alkaline digestion) leads to an over-estimation of the proportion of digestible carbohydrates calculated by difference; 2) fiber may alter the polysaccharide utilization of some foods, as shown by the "glycemic index" . It is difficult to make recommendations on dietary fiber due to insufficient data on intake, fiber composition, its physiological effects, and epidemiological studies . However, a preliminary evaluation of the diets from most Latin American countries shows large intakes of vegetable foods and, consequently, an adequate fiber intake may be expected.

Dev Biol, 1988 Sep, 129(1), 140 - 6
Characterization of genes which are transiently expressed during the preaggregative phase of development of Dictyostelium discoideum; Singleton CK et al.; We have identified and characterized three genes, the I genes (I for induced), which are induced during the preaggregative phase of the developmental program of Dictyostelium discoideum . None of these genes are expressed in cells growing vegetatively on bacteria or in axenic broth, and their induction during early development is due to transcriptional activation . Developmental expression of I6, I8, and I11 occurs even in the absence of protein synthesis . Their induction is very rapid and occurs essentially at the onset of development . The expression is transient, peaking between 2 and 4 hr followed by a rapid loss of expression . These characteristics suggest that the induction of I6, I8, and I11 is a primary result of the initiation of development, and thus they represent the first such genes isolated . Although their expression behavior shares these characteristics, examination of their expression under various conditions of development and in a variety of aggregation-deficient mutant strains reveals that the details of the regulation and developmental control of these three genes are distinct.

Biochem Biophys Res Commun, 1988 Aug 30, 155(1), 123 - 9
Intracellular pH of halobacteria can be determined by the fluorescent dye 2', 7'-bis(carboxyethyl)-5(6)-carboxyfluorescein; Tsujimoto K et al.; Determination of the internal pH of halobacterial cells grown in 4M salt solution has proven to be a difficult problem . We now report the steady state cytosolic pH of Halobacterium halobium S-9 to be 7.2 . Intracellular pH was determined after the cells were loaded with the membrane permeable precursor of the pH sensitive dye 2', 7'-bis-(carboxyethyl)-5(6)-carboxyfluorescein-acetyxymethyl ester - (BCECF/AM) . In order to minimize light-scattering in the measurement of the fluorescence, a thin cuvette was newly devised . This method should be suitable for studies of the cytosolic pH in other bacteria.

Isr J Med Sci, 1988 Aug, 24(8), 401 - 4
Acute acalculous cholecystitis--experimental and clinical observations; Shaked G et al.; Induction of acute acalculous cholecystitis was studied in dogs . Designed models included ligation of the cystic artery and/or duct separately or combined and injection of colonic flora into the gallbladder . Cholecystectomy was performed 72 h later . Histological studies of the gallbladders and bile cultures demonstrated various degrees of cholecystitis, the severest form occurring as a result of cystic artery ligation, whereas the mildest was produced by injection of bacteria . Secondary infection was a constant feature of our experimental models.

Am J Obstet Gynecol, 1988 Aug, 159(2), 363 - 8
Fetal body and breathing movements as predictors of intraamniotic infection in preterm premature rupture of membranes; Goldstein I et al.; A prospective study of fetal behavior in cases complicated with preterm premature rupture of membranes was conducted in 41 patients . The length of time and the number of fetal breathing and gross body movements were correlated with the amniotic fluid culture results . An episode of fetal activity (body movements and breathing movements) of greater than or equal to 30 seconds during 30 minutes of observation was associated with the absence of intraamniotic infection in 100% of the cases . On the other hand, the absence of fetal breathing movements and gross body movements of less than a 50 second duration during 30 minutes of observation was associated with positive amniotic fluid cultures in all cases . If an episode of fetal breathing movements was present but lasted less than 30 seconds and/or the total time of gross body movements was greater than 50 seconds, 64% of patients had an intraamniotic infection.

Infect Immun, 1988 Aug, 56(8), 2089 - 93
Influence of genetic background on host resistance to experimental murine tularemia; Anthony LS et al.; The host response to experimental murine tularemia was examined in different inbred mouse strains . The kinetics of growth of Francisella tularensis live vaccine strain (LVS) in the livers and spleens of A and C57BL/6 mice were monitored, and it was observed that mice of the A strain were more susceptible to the proliferation of LVS than were C57BL/6 mice . The difference was most marked 5 days following infection, when the number of bacteria isolated from the spleens of A mice was found to exceed that of C57BL/6 mice by 100-fold . In addition, the C57BL/6 strain exhibited a more pronounced splenomegaly 8 days after infection than did the A strain . When the response of other inbred strains was evaluated by determining the splenic count of LVS on day 5 postinfection, several levels of antiularemic resistance were observed . Mice of the AKR, BALB/cBy, C57BL/10, and SJL strains were found to be most resistant, while SM mice were most susceptible to the proliferation of LVS . The DBA/2, CBA, 129, C3H/HeJ, and A strains expressed a resistance phenotype which was intermediate between the two extremes, with A and C3H/HeJ mice being somewhat more susceptible than DBA/2, CBA, or 129 mice . The trait of resistance or susceptibility was analyzed genetically in (C57BL/6 x A)F1 hybrid mice and in F2 generation and recombinant inbred (RI) mouse strains derived from C57BL/6 (resistant) and A (susceptible) strain progenitors . The F1 progeny exhibited a level of resistance to infection which was similar to that of the resistant parent . In both the F2 generation mice and the RI strains, a continuous spectrum of resistance levels was observed . The results of these experiments indicate that the genetic background of the host influences host resistance to experimental murine tularemia and that multiple genetic loci are involved in this response.

Infect Immun, 1988 Aug, 56(8), 1890 - 6
Ultrastructural localization of capsules, cell wall polysaccharide, cell wall proteins, and F antigen in pneumococci; Skov Sorensen UB et al.; The localization of pneumococcal capsular and cell wall antigens was examined by immunoelectron microscopy . C polysaccharide (C-Ps), a common component of all pneumococci, was uniformly distributed on both the inside and outside of the cell walls . The thickness of the C-Ps varied with the strain . Encapsulated strains were covered by varied amounts of capsular polysaccharide concealing the C-Ps of the bacteria so as to render it inaccessible to anti-C-Ps antibodies . In addition to C-Ps, protein antigens were demonstrable on the surface of nonencapsulated pneumococci . The proteins were not masked by the C-Ps layer . An extra layer on the cell walls was conspicuous on electron micrographs of both rough and encapsulated pneumococci . The nature of this extra layer has not been disclosed . F antigen, another common antigen of pneumococci, was uniformly distributed on the surface of the plasma membranes . During the course of the experimental work a reproducible method of gold labeling immunoglobulins was developed.

Chest, 1988 Aug, 94(2 Suppl), 126S - 129S
Comparison of two nonculture techniques for detection of Hemophilus influenzae in sputum . In situ hybridization and immunoperoxidase staining with monoclonal antibodies; Terpstra WJ et al.; Two nonculture methods, in situ hybridization and immunoperoxidase staining with monoclonal antibodies, were compared for the detection of Hemophilus influenzae in 184 sputa . For in situ hybridization, a biotin-labeled probe of total genomic DNA of H influenzae type b was prepared that hybridizes specifically with H influenzae, H parainfluenzae, H hemolyticus, and H parahemolyticus DNA . Immunoperoxidase staining was done with monoclonal antibody 8BD9 directed against outer membrane protein P6 of H influenzae . Both techniques detected Hemophilus in sputum equally well and were superior to culture: all 30 sputum samples culture-positive for H influenzae were positive on both nonculture tests, and 13 additional positive sputum samples were detected from which Hemophilus was not cultured . The higher sensitivity of the nonculture tests was mainly attributed to culture failure because of overgrowth of H influenzae by other bacteria, especially in patients with cystic fibrosis . The immunoperoxidase staining technique appeared slightly easier and quicker to perform than the in situ hybridization test . For the in situ DNA hybridization probe, DNA can be prepared from any strain of H influenzae . The immunoperoxidase test requires monoclonal antibody 8BD9 but has a higher specificity than the hybridization technique . Both techniques can be reliably applied, especially for the detection of Hemophilus in sputum of patients with cystic fibrosis.

Int J Food Microbiol, 1988 Aug, 7(1), 1 - 7
The effect of lactic acid sprays on the keeping qualities of meat during storage; Cudjoe KS; Spraying the meat surface of skinned cow heads with 1% v/v lactic acid resulted in significant reduction in total viable counts of bacteria at storage temperatures of 4, 15 and 20 degrees C . The number of coliform bacteria was also reduced at all three temperatures but the reductions were not statistically significant on most occasions . However, after five and two days at 15 degrees C and 20 degrees C, respectively, when the initial effect of acid appeared to be lost, the number of coliforms on sprayed heads exceeded those on unsprayed heads . The shelf lives of all sprayed heads were observed to have been extended for about three days at 4 degrees C and one day for both 15 degrees C and 20 degrees C.

Br J Rheumatol, 1988 Aug, 27(4), 306 - 9
The role of HLA-B27 in arthritis; Archer JR et al.; We postulate (Table I) that ReA is an antigen or immune complex induced condition caused by chronic intracellular bacterial infection at a distant site . The main predisposing factor is a failure to resist this infection . If the bacteria happen to carry MF, the inflammation is exacerbated in B27 positive patients . In contrast AS occurs in individuals who lack immunity to MF and eventually become infected by an intracellular organism which synthesizes it (virus or plasmid?) . HLA-B27 acts only at the site of inflammation.

Infect Immun, 1988 Aug, 56(8), 2028 - 35
Chemical and biological characterization of the lipopolysaccharide of the oral pathogen Wolinella recta ATCC 33238; Gillespie J et al.; To investigate the potential pathogenic mechanisms of the oral periodontopathogen Wolinella recta ATCC 33238, we have isolated its lipopolysaccharide (LPS) and determined the chemical composition and selected in vitro biological activities of the molecule . Sodium desoxycholate-polyacrylamide gel electrophoresis revealed the W . recta LPS to be an atypical smooth LPS with short O-antigenic side chains . Chemically the LPS consisted of 47.2% lipid A, 19.6% polysaccharide, 9.0% heptose, 8.5% hexosamine, 3.2% phosphate, and 0.6% 2-keto-3-deoxyoctanoate . The major fatty acids were hexadecanoic acid (25.0%), 3-OH tetradecanoic acid (23.8%), tetradecanoic acid (15.4%), 3-OH hexadecanoic acid (11.6%), and octadecenoic acid (10.9%) . Rhamnose constituted 87.8% of the carbohydrates generally associated with the O antigen, with smaller amounts of glucose (5.5%), mannose (4.9%), and an unidentified sugar (1.9%) . CD-1 and C3H/HeN macrophages (M phi) exposed to 1 microgram of W . recta LPS per ml released 6.0 and 10.5 ng of prostaglandin E per ml of supernatant, representing 625% and 1,306% of prostaglandin E release by the control (without LPS) . Maximum prostaglandin E release occurred in CD-1 M phi exposed to 100 micrograms of LPS per ml and was equivalent to 1,542% of release by the control . Interleukin-1 (IL-1) activities in CD-1 and C3H/HeN M phi exposed to 1 micrograms of LPS per ml were 257% and 1,941% of activities in the control, respectively . Maximum IL-1 release in CD-1 M phi occurred in response to 50 micrograms of LPS per ml and represented a 927% increase over release in the control, while 100 micrograms LPS per ml stimulated maximum IL-1 release in C3H/HeN M phi that was greater than 5,000% of release by the control.

J Gen Microbiol, 1988 Aug, 134 ( Pt 8), 2149 - 54
Utilization of histidine by Caulobacter crescentus; Ferber DM et al.; Caulobacter crescentus has an inducible pathway which is responsible for the degradation of histidine . Induction of this pathway occurs in the presence of both glucose and ammonia . Growth yield experiments indicate that only two of the three available nitrogens are used for growth suggesting that formamide may be produced as a waste product . However, formamide was not detected in the culture fluid and formate was formed instead . These results suggest that histidine may be degraded in a novel pathway which results in the production of 1 mol each of ammonia, glutamate and formate per mol of histidine . The third nitrogen from histidine appears to be sequestered in some kind of secondary metabolite.

Pharmazie, 1988 Aug, 43(8), 537 - 8
Synthesis and biological activity of some new pyrimidine and thienopyrimidine derivatives; el-Bahaie S et al.; The readily available 4-mercapto-6-methyl-2-phenyl-5-acetylpyrimidine (1) was alkylated with a variety of activated halomethylene compounds to give the corresponding thieno {2,3-d} pyrimidine derivatives 3a-h via the intermediate alkylmercaptopyrimidine derivatives 2b-h . The biological activity of the new compounds was screened against several strains of bacteria.

Microb Pathog, 1988 Aug, 5(2), 87 - 95
Growth of Legionella pneumophila in a human macrophage-like (U937) cell line; Pearlman E et al.; We established a model of the bacteria-macrophage interaction to study the cellular basis of Legionella pneumophila pathogenesis and to characterize avirulent L . pneumophila . We found that U937 cells, which are derived from a human histiocytic lymphoma cell line, support intracellular growth of L . pneumophila with a doubling time of 6 h, and that sustained intracellular growth is associated with a cytopathic effect (CPE) that can be detected by microscopic examination and quantified with the vital stain 3-(4,5-dimethyl thiazol-2-yl)-2,5,-diphenyl tetrazolium bromide (MTT) . An L . pneumophila isolate obtained directly from infected guinea-pig spleens can grow and produce CPE in these cells, destroying most of the cell layer after 72 h of growth . Only 10(6) organisms of this strain are required to kill 50% of guinea-pigs inoculated by the intraperitoneal route . In contrast, an avirulent isolate derived by 203 successive plate passages of the same strain can neither kill guinea-pigs at an intraperitoneal inoculum of 10(7) nor grow or produce CPE in U937 cells . Since the cells were able to differentiate between a virulent and an avirulent strain of L . pneumophila, we conclude that U937 cells are an appropriate model system for study of the bacteria-macrophage interaction.

Genomics, 1988 Aug, 3(2), 137 - 42
Hypotheses for testing deviations from random integration: evidence for nonrandom retroviral integration; Wilson AF et al.; The use of a recently developed in vitro model for retroviral integration provides a means of statistically testing hypotheses concerning the distribution of integration sites and hypotheses about the sequence of proviral orientations . In this study, three null hypotheses are formulated and applied to previously published data . Statistical analyses of these data suggest that the distribution of integration sites may not be uniform, and the sequence of proviral orientations is not random . On the basis of these results and the observed clustering of orientations, it was postulated that if a DNA sequence was involved in nonrandom proviral integration, that sequence would be found in the regions where the orientations change direction with respect to the target DNA ("I" regions) . Computer analyses for homologous and complementary DNA sequences were performed on all possible pairs of identifiable "I" regions . A common sequence (at least 8 bp in size) was found in three out of four regions and that sequence was absent elsewhere in the target DNA . A model, with features of recombination reminiscent of chi sequences in bacteria, is proposed that may account for these results.

Burns Incl Therm Inj, 1988 Aug, 14(4), 297 - 302
A model to reproduce predictable full-thickness burns in an experimental animal; Kistler D et al.; A standardized, reproducible animal model is a prerequisite to study concepts in the therapy of extensive burn injuries . The development of a new model makes it possible to produce predetermined burn injuries with a set temperature, time, contact pressure, and standard extent of tissue damage . For our studies we chose rats and exposed them to a temperature of 250 degrees C for 20 s and a contact pressure of 500 g/cm2 over various percentages of TBSA (total body surface area) . The animals received shock prophylaxis for 3 days postburn and were kept under standardized conditions in a laminar airflow compartment . The temperature was kept at 32 degrees C and the relative humidity at 75 per cent . To reduce bacterial contamination, air was filtered through special bacteria-proof filters . Under these conditions we found burns of approximately 35 per cent TBSA to be sublethal resulting in 80 per cent mortality between days 5 and 7 . This model permits the investigator to vary the burned skin area to any required extent for a reproducible study of different concepts of burn therapy.

Chemioterapia, 1988 Aug, 7(4), 253 - 5
The treatment of gynaecological and intra-abdominal infections: a comparative study of cefotetan versus netilmicin plus clindamycin; Poularas J et al.; In a prospective randomised study 60 patients with gynaecological or intra-abdominal infections were given either 2g iv every 12 hours of cefotetan or a combination of netilmicin (150mg iv every 12 hours) and clindamycin (600mg iv every 8 hours) . The clinical condition of nearly half the patients (26 of 60) was characterized as serious and surgical manipulation and drainage were performed in 57 of the 60 patients . The clinical response was similar in both groups with 21 of 29 patients in the cefotetan group and 29 of 33 patients in the netilmicin plus clindamycin group . Side effects were few and mild in nature with no significant differences between the two groups . This work is continuing but the results to date suggest that cefotetan monotherapy is a safe and effective alternative to a combination of netilmicin and clindamycin in the treatment of gynaecological and intra-abdominal infections.

Appl Environ Microbiol, 1988 Aug, 54(8), 2003 - 9
Effects of acidification on mercury methylation, demethylation, and volatilization in sediments from an acid-susceptible lake; Steffan RJ et al.; The effect of experimental acidification on mercury methylation, demethylation, and volatilization was examined in surficial sediment samples from a weakly buffered northern Wisconsin lake . All mercury transformations were measured with radioisotopic tracers . Acidification of sediment pH with H2SO4, HCl, or HNO3 significantly decreased 203Hg(II) methylation . Acidification of pH 6.1 (ambient) sediments to pH 4.5 with either H2SO4 or HCl inhibited methylation by over 65% . The decreased methylation was due to the increased hydrogen ion concentration because methylation was not affected by concentrations of Na2SO4 or NaCl equimolar to the amount of acid added . Inhibition of methylation was observed even after prolonged acidification of sediments to pH 5.0 for up to 74 days . Acidification of sediments to pH 5.5, 4.5, and 3.5 with HNO3 resulted in a near complete inhibition of methylation at each pH . Similarly, the addition of equimolar amounts of NaNO3 resulted in a near complete inhibition of methylation, indicating that the inhibition was due to the nitrate ion rather than to the acidity . Demethylation of methyl mercury was not affected by pHs between 8.0 and 4.4, but sharply decreased below pH 4.4 . Volatilization of 203Hg(II) from surface sediments was less than 2% of methylation activity and was not significantly different from that in killed sediments . This study indicated that acidification of sediments inhibits mercury methylation and that the observed increase in the mercury burdens in fish from low pH lakes is not due to increased production of methylmercury in sediments.

Jpn J Antibiot, 1988 Aug, 41(8), 997 - 1001
{Transfer of cefuzonam into female genital organs}; Hongo M et al.; Cefuzonam (CZON) was given intravenously at a 1 gram dose to 24 patients prior to abdominal total hysterectomy for uterine myoma . Concentrations of CZON in the serum of cubital vein, uterine artery, and in tissues of uterus and adnexa uteri were measured by the bioassay method . Bilateral uterine arteries were clamped at 15 and 30 minutes and 1, 2, 4 and 6 hours after the administrations of the drug . Measurements were done on 4 patients at each sampling . Concentrations of CZON in the serum of cubital vein and uterine artery changed in a similar manner with respective half-lives of 52.0 minutes and 48.2 minutes . Peak concentrations in oviduct, ovary, endometrium, myometrium, cervix uteri and portio vaginalis were reached at 15 minutes after drug administration . The peak concentration in the ovary was as high as 37.5 micrograms/g, and those in other tissues varied between 17.6 and 25.5 micrograms/g . Tissue concentrations decreased gradually after their peaks in similar manners to those in serum, and their half-lives ranged between 37.4 and 53.7 minutes . From these results, it was found that CZON was transferred to female genital organs in high enough concentrations to inhibit growth of bacteria which were often isolated from the pelvic inflammatory diseases, and CZON seemed to be highly effective to clinical pelvic inflammatory diseases.

J Prosthet Dent, 1988 Aug, 60(2), 212 - 8
Effects of sodium hypochlorite on denture base metals during immersion for short-term sterilization; McGowan MJ et al.; The effects on Vitallium and Ticonium alloys immersed in different concentrations of sodium hypochlorite for various immersion times are described . The results of this investigation demonstrate that samples of these metals soaked in 2% sodium hypochlorite for at least 5 minutes or in 5.25% sodium hypochlorite for a minimum of 3 minutes were sterilized and not tarnished or corroded . Although Vitallium alloy was more resistant than Ticonium alloy to the deleterious effects of sodium hypochlorite, either metal can be safely immersed for sterilization purposes under the guidelines presented . Therefore, an effective and economical method for sterilizing removable partial denture frameworks made of chrome-cobalt or chrome-nickel alloys is attainable through the proper use of household bleach.

Genitourin Med, 1988 Aug, 64(4), 273 - 5
Rectal occurrence of Mobiluncus species; Hallen A et al.; The simultaneous occurrence of Mobiluncus spp in the vagina and rectum was studied in women attending a sexually transmitted diseases (STD) department . Of 43 women with bacterial vaginosis (BV), 32 had Mobiluncus spp in the vagina and 23 in the rectum . In 20 women the same Mobiluncus species was found concomitantly in the vagina and the rectum . Mobiluncus spp were found in the rectum only in two women with BV, one with candidal vaginitis, and one healthy woman . In a treatment study of 23 women, BV was cured in nine out of 10 treated with metronidazole and six out of 13 treated with tetracycline . After treatment Mobiluncus spp persisted in the vaginas of two out of 19 women and in the rectums of two out of 10.

Clin Invest Med, 1988 Aug, 11(4), 297 - 303
Experimental respiratory infection with Bordetella pertussis in mice: comparison of two methods; Halperin SA et al.; The mouse respiratory model is being used increasingly to study the pathogenesis and immunology of Bordetella pertussis infection . Two methods of inoculation, aerosol and intranasal, are routinely used to establish the infection . We compared the two methods of inoculation for reproducibility of infection using quantitative lung cultures and distribution of infection with {35S} methionine labeled bacteria and pulmonary histopathology . Ability to produce a respiratory infection intranasally was related to the inoculum volume; a minimum of 20 microliters was required although considerable variability remained . Lung bacterial counts in identically inoculated mice varied 1,000 fold following intranasal inoculation compared to only 5 fold following aerosol inoculation . Distribution of pulmonary 35S-labeled bacteria varied widely (right lung, 43-84%; left lung 16-57%) following intranasal in comparison to aerosol inoculation (right, 60-68%; left 32-40%) . Finally, intranasal inoculation produced a scant, patchy, bronchopneumonia whereas diffuse pathology involving all pulmonary segments was seen following aerosol infection . Due to the superior reproducibility and predictable distribution of infection and pathology, aerosol inoculation is the method of choice for establishing the mouse model of pertussis respiratory infection.

J Electron Microsc Tech, 1988 Aug, 9(4), 413 - 9
Correlation averaging of a badly distorted lattice: the surface protein of Pyrodictium occultum; Hegerl R et al.; The surface protein of the archaebacterium Pyrodictium occultum forms two-dimensional periodic arrays of extremely poor order . Two variants of correlation averaging have been applied in order to retrieve the unit cell structure from electron micrographs of negatively stained samples: straightforward correlation averaging correcting for lateral displacements only and a more elaborate approach, including a partial compensation for rotational disorder . Surprisingly, both routes yield virtually identical structures . Inclusion of molecular motifs from highly disordered domains, which are rejected in the "straightforward" approach, appears not to improve resolution, possibly because the high local strain tends to distort the individual molecules.

Burns Incl Therm Inj, 1988 Aug, 14(4), 326 - 30
Preliminary clinical studies of a biological skin equivalent in burned patients; Wassermann D et al.; The possibility of covering large areas of full thickness skin loss with 'living skin equivalent' produced by a modification of Bell's method was studied . Living skin equivalents, composed of a dermal equivalent (fibroblasts plus collagen) covered by epithelial cells were grafted, meshed or non-meshed, onto granulation tissue and, in one patient, onto fascia . Eight patients with full skin thickness burn wounds covering over 15 per cent of the body surface area were thus partially covered . The graft 'take' was evaluated every 48 h . In every patient grafted, an extensive lysis (60-90 per cent) of the skin equivalent graft was observed at the first dressing (48 h) . In one patient only, a significant percentage of 'take' (40 per cent) was observed 14 days after grafting . These disappointing results were probably related to the presence of collagenases or proteases produced on the wound bed either by bacteria or by surrounding human cells . It appears that at the present time the biochemical nature of the dermal equivalent used is not yet completely appropriate to serve routinely as a substitute for human skin.

J Electron Microsc Tech, 1988 Aug, 9(4), 301 - 24
Introduction to the computer image processing of electron micrographs of two-dimensionally ordered biological structures; Stewart M; Methods are described for the analysis of electron micrographs of regular biological objects . Fourier-based processing of one-dimensionally ordered arrays is described by way of introduction, before analysing two-dimensional crystals in projection with the aim of enhancing signal:noise ratio and thus of feint features that were initially obscured . This form of analysis is then extended to decomposing the moire patterns formed when sheets overlap, thereby enabling the separation of interfering image patterns . Analogous forms of an analysis can also be applied to objects with rotational symmetry . Methods for treating the effect of the microscope imaging system and compensating for lattice disorder in crystalline specimens are also discussed.

J Protozool, 1988 Aug, 35(3), 389 - 93
Malaria sporozoites leave behind trails of circumsporozoite protein during gliding motility; Stewart MJ et al.; As Plasmodium sporozoites undergo gliding motility in vitro, they leave behind trails of circumsporozoite (CS) protein that correspond to their patterns of movement . This light microscopic observation was made using Plasmodium berghei sporozoites, a monoclonal antibody (MAb H4) directed against the immunodominant repetitive epitope of the CS protein of P . berghei, and an immunogold-silver staining (IGSS) technique . Sporozoites pretreated with agents that inhibit sporozoite motility and invasiveness did not produce trails . Sporozoites that glided on microscope slides coated with MAb H4 left behind considerably longer CS protein trails than those on uncoated slides, and the staining of these trails was more intense . The fact that the CS protein is an exoantigen continuously released as trails by motile sporozoites, together with our previous finding that anti-CS protein antibodies inhibit sporozoite motility, strongly suggests that the CS protein plays a role in gliding motility . The sensitive IGSS technique used in this study may be a useful tool in the study of the translocation of surface proteins during gliding of other apicomplexans, other protists, and bacteria.

Ecotoxicol Environ Saf, 1988 Aug, 16(1), 65 - 71
Review of the environmental toxicity of quaternary ammonium halides; Cooper JC; This paper summarizes all available information on the environmental toxicity of quaternary ammonium compounds (QACs) . Ten-minute contact kills of bacteria occur at 50-333 mg/liter . These chemicals are acutely toxic at approximately 1 mg/liter to invertebrates and fish and toxicity is occasionally as low as 0.1 mg/liter; no-effect levels are generally 10 times lower than LC50 values . Toxicity to invertebrates and fish appears to be relatively independent of structure; the compounds studied, which have a large variety of structures, are all toxic at approximately the same order of magnitude . The QACs tested inhibited plant growth at 3-5 mg/liter . Predictions of toxicity to QACs are made based on knowledge of structure-activity relationships.

Vet Clin North Am Equine Pract, 1988 Aug, 4(2), 221 - 8
Uterine defense mechanisms in the mare; Liu IK; Uterine defense against infection in the mare has been actively investigated over the past decade . Mechanisms of defense, including the role of immunoglobulins, polymorphonuclear neutrophils, and the physical ability of mares to eliminate bacteria from the uterus, are discussed.

J Gen Microbiol, 1988 Aug, 134 ( Pt 8), 2363 - 74
Identification of species of the genus Legionella using a cloned rRNA gene from Legionella pneumophila; Saunders NA et al.; A cloned EcoRI fragment from Legionella pneumophila, which includes 16S and 23S rRNA genes, was used to identify bacteria belonging to the genus Legionella by hybridization to a series of species specific restriction fragments . Examination of the type strains of 28 species of legionellae gave different band patterns in every case . When further isolates of these species were tested the patterns obtained were usually either identical, or very similar, to those of the respective type strains . Thirty-one coded isolates were examined and of these 29 were allocated to the correct species . The remaining strains (a non-Legionella and a L . pneumophila) could not be identified using this technique . The rRNA gene probe method should be of great value in the identification of legionellae, particularly for those species which are at present very difficult to distinguish serologically.

Gene, 1988 Jul 30, 67(2), 271 - 7
A simple phase-extraction assay for chloramphenicol acyltransferase activity; Seed B et al.; A simple and convenient phase extraction assay for chloramphenicol (Cm) acetyltransferase (CAT) activity has been developed, based on the enzymatic butyrylation of radiolabelled Cm . The assay is linear over two to three orders of magnitude of enzyme concentration, is highly sensitive, and substantially less expensive than all presently available alternatives . Methods for convenient CAT assay, adapted for mammalian cells, plant protoplasts, and mammalian cell culture supernatants, are described . These methods should also simplify measurement of CAT activity in other organisms, such as yeast and bacteria . In addition, a simple pre-extraction procedure is presented for purifying radiolabelled Cm which allows a 25-fold increase in sensitivity using tritiated substrates.

Nucleic Acids Res, 1988 Jul 25, 16(14B), 6987 - 99
Oligonucleotide-directed construction of mutations: a gapped duplex DNA procedure without enzymatic reactions in vitro; Fritz HJ et al.; The gapped duplex DNA approach to oligonucleotide-directed construction of mutations (Kramer et al . 1984, Nucl . Acids Res . 12, 9441-9456) has been developed further . A procedure is described that makes in vitro DNA polymerase/DNA ligase reactions dispensable . Direct transfection of host bacteria with gdDNA molecules of recombinant phage M13 plus mutagenic oligonucleotide results in marker yields in excess of 50% (gap size 1640 nucleotides) . An important feature incorporated into the mutagenic oligonucleotide is the presence of one or two internucleotidic phosphorothioate linkages immediately adjacent to the 5'-terminus . Automated preparation and biochemical properties of such compounds are described as well as their performance in oligonucleotide-directed mutagenesis . A systematic study of the following parameters influencing marker yield is reported: Gap size, length of oligonucleotide, chemical nature of oligonucleotide termini and heatshock temperature during transformation.

J Biol Chem, 1988 Jul 25, 263(21), 10048 - 51
RNA polymerases B and C are more closely related to each other than to RNA polymerase A; Memet S et al.; Amino acid sequence comparison of the largest subunit of the three forms of yeast nuclear RNA polymerase disclosed six major conserved regions that are partly retained in the cognate subunits from bacteria, viral, and insect enzymes (Memet, S., Gouy, M., Marck, C., Sentenac, A., and Buhler, J.-M . (1988) J . Biol . Chem . 263, 2830-2839) . Within these conserved domains, the high sequence similarity of B220 and C160 subunits (52% identity) sets them apart from yeast enzyme A subunit A190 . Parsimony analysis at the gene and protein levels suggests the existence of a transient ancestor to eukaryotic RNA polymerases B and C . These results are discussed in the light of the recent finding of class C genes containing RNA polymerase B promoter elements.

Biochim Biophys Acta, 1988 Jul 13, 950(2), 193 - 203
Isolation, characterization and microsequence analysis of a small basic methylated DNA-binding protein from the Archaebacterium, Sulfolobus solfataricus; Choli T et al.; DNA-binding proteins have been extracted from the thermoacidophilic archaebacterium Sulfolobus solfataricus strain P1, grown at 86 degrees C and pH 4.5 . These proteins, which may have a histone-like function, were isolated and purified under standard, non-denaturing conditions, and can be grouped into three molecular mass classes of 7, 8 and 10 kDa . We have purified to homogenity the main 7 kDa protein and determined its DNA-binding affinity by filter binding assays and electron microscopy . The Stokes radius of gyration indicates that the protein occurs as a monomer . The complete amino-acid sequence of this protein contains 14 lysine residues out of 63 amino acids and the calculated Mr is 7149 . Five of the lysine residues are partially monomethylated to varying extents and the methylated residues are located exclusively in the N-terminal (positions 4 and 6) and the C-terminal (positions 60, 62 and 63) regions only . The protein is strongly homologous to the 7 kDa proteins of Sulfolobus acidocaldarius with the highest homology to protein 7d . Accordingly, the name of this protein from S . solfataricus was assigned as DNA-binding protein Sso7d.

Jikken Dobutsu, 1988 Jul, 37(3), 311 - 6
Enhancing effect of phenobarbital sodium on Tyzzer's disease in mice; Xian M et al.; Tyzzer's disease in mice was aggravated by phenobarbital sodium (PB) given consecutively after bacterial inoculation . In PB-treated mice, mortality rate and severity of liver lesions were higher with more prominent bacterial propagation in hepatocytes as compared with non-treated ones, suggesting that PB had an enhancing effect on metabolic activities of host hepatocytes resulting in increased intracellular growth of bacteria.

Hum Toxicol, 1988 Jul, 7(4), 337 - 41
Studies of the 'adaptive' repair response in human lymphocytes and V79 cells after treatment with MNNG and MNU; Anderson D et al.; In bacteria, there is evidence that a damage inducible repair response system known as the adaptive response exists since pretreatment with low doses of a simple monofunctional alkylating agent leads to a decrease in both the lethal and mutagenic effects of a subsequent challenge dose of the agent . The evidence for an analogous system in mammalian cells has proved to be inconsistent to date . The induction of chromosome repair mechanisms in human cells by low-dose radiation from tritiated thymidine has been shown to make the cells refractory to the induction of chromosome aberrations by X-rays . The present communication investigates the induction of an adaptive response in human lymphocytes from four donors and V79 cells using SCE and mutation as endpoints and MNNG and MNU for the adapting and challenging treatment . It is clear that a reproducible model of the adaptive response in human lymphocytes is difficult to establish because of the variability between different donors and different culture times . In V79 cells, assays with much larger cell numbers are required to detect a reproducible response with such small changes in mutant frequency . To demonstrate an adaptive response conclusively in mammalian cells will probably require the use of more sensitive experimental protocols and alternative methods of administration of adaptive doses of mutagen.

Br J Nutr, 1988 Jul, 60(1), 185 - 92
Degradation of hemicellulose and pectin by horse caecum contents; Bonhomme-Florentin A; 1 . Polysaccharide depolymerases and glycoside hydrolases involved in the breakdown of plant structural polysaccharides (hemicellulose and pectins) were monitored in three fractions of the liquid phase of horse caecum digesta: acellular fluid (AF), bacteria (B) and protozoa plus bacteria (PB) . 2 . Both bacteria and protozoa were found to be involved in the decomposition of pectic substances, with two enzymic activities: depolymerase (polygalacturonase, EC 3.2.1.15; and pectin lyase, EC 4.2.2.10) and esterase (pectinesterase, EC 3.1.1.11) . The activity of the PB fraction was higher than that of B . 3 . With hemicellulosic substrates, all three fractions showed a significant xylan endo-1,3-beta-xylosidase (EC 3.2.1.32) activity . Mannan was hardly broken down . 4 . Galactomannan and arabinogalactan were broken down more extensively by the PB fraction than by the B fraction . Glycosidase activities (xylan 1,4-beta-xylosidase, EC 3.2.1.37 and alpha-L-arabinofuranosidase, EC 3.2.1.55) were also observed.

J Bacteriol, 1988 Jul, 170(7), 3189 - 93
Purification and properties of formate dehydrogenase from Moraxella sp . strain C-1; Asano Y et al.; NAD+-dependent formate dehydrogenase was screened in various bacterial strains . Facultative methanol-utilizing bacteria isolated from soil samples, acclimated to a medium containing methanol and formate at pH 9.5, were classified as members of the genus Moraxella . From a crude extract of Moraxella sp . strain C-1, formate dehydrogenase was purified to homogeneity, as judged by disc gel electrophoresis . The enzyme has an isoelectric point of 3.9 and a molecular weight of approximately 98,000 . The enzyme is composed of two identical subunits with molecular weights of about 48,000 . The apparent Km values for sodium formate and NAD+ were calculated to be 13 mM and 0.068 mM, respectively.

Gastroenterology, 1988 Jul, 95(1), 164 - 9
Kinetics for the synthetic bile acid 75selenohomocholic acid-taurine in humans: comparison with {14C}taurocholate; Jazrawi RP et al.; The "apparent" fractional turnover rate of the gamma-labeled bile acid analogue 75selenohomocholic acid-taurine (75SeHCAT) was assessed from decline in radioactivity over the gallbladder area on 4 successive days using a gamma-camera, and was compared in the same subjects with the fractional turnover rate of the corresponding natural bile acid, cholic acid-taurine, labeled with 14C ({14C}CAT) using the classical Lindstedt technique . Very similar results were obtained in 5 healthy individuals (coefficient of variation 4.8%, medians 0.35 and 0.34, respectively) . By contrast, the fractional deconjugation rate assessed from zonal scanning of glycine- and taurine-conjugated bile acids on thin-layer chromatography was much less for 75SeHCAT than for {14C}CAT (0.02 and 0.13, respectively; p less than 0.05) . The fractional rate for deconjugation plus dehydroxylation was also determined by zonal scanning, and gave lower values for 75SeHCAT than for {14C}CAT (0.02 and 0.12, respectively; p less than 0.05) . There was a striking similarity between the fractional rate for deconjugation alone and that for deconjugation plus dehydroxylation for both bile acids in individual samples (r = 0.999, p less than 0.001), suggesting that these two processes might occur simultaneously and probably involve the same bacteria . We conclude that our scintiscanning technique provides an accurate, noninvasive method of measuring fractional turnover rate of a bile acid in humans, and that the finding that 75SeHCAT remains conjugated with taurine during enterohepatic recycling means that absorption should be specific for the ileal active transport site, thus rendering it an ideal substance for assessing ileal function.

Eur J Biochem, 1988 Jul 1, 174(4), 621 - 7
Conformational alterations detected by circular dichroism induced in the normal ras p21 protein by activating point mutations at position 12, 59, or 61; Valencia A et al.; Activation of the oncogenic potential of ras oncogenes occurs by point mutations at codons 12, 13, 59, 61, and 63 of the sequences that codify for its product, a 21-kDa protein designated as p21 . This activation has been postulated by computer models as modifiers of the structure of the protein, which may alter its biochemical and biological activities . We have expressed in bacteria the normal ras p21 and five mutated p21 proteins with mutations at positions 12, 59, 61, 12 plus 59, and 12 plus 61 . Purification was carried out by solubilization from bacterial pellets in 7 M urea and chromatography through a Sephadex G-100 column to obtain greater than 95% purified proteins . Circular dichroic (CD) spectra showed that the normal protein and that activated by substitution of Ala59 to Thr59 are very similar in their overall structure . By contrast, point mutations affecting either 12 or 61 residues substantially altered the structure of the proteins . When the parameters of Chen et al . {Biochemistry II, 4120-4131 (1972)} were applied to the CD spectra, both normal and thr59-mutated ras proteins showed a less organized structure than mutated proteins at position 12 or 61 . Since the Thr59 mutant has more similar transforming activity than other activated proteins, but a GTPase activity similar to that of the normal protein, our results support the hypothesis that there is more than one mechanism of activation of the ras p21 protein . One of these mechanisms involves important structural alterations by point mutations at position 12 or 61 which reduce the GTPase activity of the protein . Another mechanism will be that induced by a substitution of Ala59 to Thr59 which does not substantially alter the protein conformation . A putative alternative mechanism for the activation of this mutant is discussed.

ASAIO Trans, 1988 Jul-Sep, 34(3), 765 - 8
Clinical experience with a new hemodiafiltration (HDF) system; Rindi P et al.; Purity of reinfusion fluid and dialysate is mandatory in high flux, high efficiency substitutive treatments for chronic uremia, particularly when using bicarbonate concentrates . Bags of sterile and pyrogen free substitution fluid are in part responsible for the high costs of these treatments . In order to overcome these problems, the authors tested a new monitoring system (MPS-10, Gambro) for high efficiency hemodiafiltration (HDF) in which the reinfusion fluid was prepared from bicarbonate dialysate by filtration . During 5 months time, 96 on-line HDF treatments were performed in five chronic uremic patients . We found the infusion fluid and dialysate containing bicarbonate free from bacteria and endotoxins, with a correct weight balance obtained by the monitoring system . The authors believe that MPS-10 is a safe and adequate system for the production of sterile, pyrogen free dialysate and reinfusion fluid for on-line HDF, hemodialysis (HD), and hemofiltration (HF).

ASAIO Trans, 1988 Jul-Sep, 34(3), 205 - 7
Easy production of sterile, pyrogen-free dialysate; Erley CM et al.; Hemodialysis (HD) hypotension is frequently encountered during conventional acetate HD . Recently it has been suggested that monocytes that adhere to the dialysis membrane are also stimulated by endotoxins diffusing from the dialysate side . Stimulated monocytes, however, release interleukin-1, which mediates fever and hypotension through its action on the cyclo-oxygenase cascade . To prevent this endotoxin-induced stimulation of monocytes, a hemofilter with a polyamide membrane (FH 88, Gambro, Lund, Sweden, cut-off 20,000 daltons, surface area 2.0 m2) was interposed between the dialysate outlet of the HD machine and the dialyzer . The data obtained clearly show that the filtered dialysate was always pyrogen-free when tested with a limulus-amebocyte-lysate assay . In addition, in 80% of cases no bacteria were detected after the sterilizing filter . Almost no febrile episodes were observed when sterile dialysate was used.

Avian Dis, 1988 Jul-Sep, 32(3), 445 - 50
Intestinal and bursal cryptosporidiosis in turkeys following inoculation with Cryptosporidium sp . isolated from commercial poults; Bermudez AJ et al.; Cryptosporidium meleagridis oocysts, originally isolated from droppings of commercial turkey poults with increased mortality due to viral (reovirus) hepatitis and enteritis, were treated with peracetic acid to kill companion bacteria and viruses and then propagated by passage in young turkeys . Thirty-eight 5-day-old large white turkey poults were inoculated by crop gavage with 500,000 cryptosporidial oocysts and compared with 40 uninoculated poults . Cryptosporidial oocysts shedding began 3 days postinoculation (PI), peaked on day 4 PI, and persisted at a low level for the duration of the 21-day trial . Low to moderate cryptosporidial infections of the ileal mucosa (days 3, 6, and 15 PI), cecal mucosa (days 3, 6, and 21 PI), and bursa of Fabricius (days 6, 12, 15 and 21 PI) were found on histopathological examination . There were no differences in mean body weights between the inoculated and uninoculated groups, and no mortality or clinical signs of disease were seen in either group.

Somat Cell Mol Genet, 1988 Jul, 14(4), 351 - 7
Enhancement of DNA repair capacity of mammalian cells by carcinogen treatment; Protic M et al.; To determine whether DNA excision repair is enhanced in mammalian cells in response to DNA damage, as it is in bacteria as part of the SOS response, we used an expression vector-host cell reactivation assay to measure cellular DNA repair capacity . When UV-damaged chloramphenicol acetyltransferase (CAT) vector DNA was introduced into monkey cells (CV-1), the level of CAT activity was inversely related to the UV fluence due to inhibition of CAT gene expression by UV photoproducts . When CV-1 cells were treated with either UV radiation or mitomycin C, 24-48 h before transfection, CAT expression from the UV-irradiated plasmid was increased . This increase also occurred in a line of normal human cells, but not in repair-deficient human xeroderma pigmentosum cells . We confirmed that this increase in CAT expression was due to repair, and not to production of damage-free templates by recombination; the frequency of generation of supF+ recombinants after transfection with UV-irradiated pZ189 vectors carrying different point mutations in the supF gene did not significantly increase in carcinogen-treated CV-1 cells . From these results we conclude that carcinogen treatment enhances the excision-repair capacity of normal mammalian cells.

Orthop Clin North Am, 1988 Jul, 19(3), 605 - 10
Role of gentamicin-impregnated cement in total joint arthroplasty; Garvin KL et al.; Palacos-Gentamicin (PG) was used in 130 joint arthroplasties at The Hospital for Special Surgery . At an average follow-up of 2.5 years (range, 2 to 5), the overall recurrence rate of infection was 3.8 per cent . Only one (2.0 per cent) infection occurred where the bacteria were sensitive to PG and there were no infections in the two-stage reimplantation group . There have been no adverse effects from the gentamicin and the inferior intrusion properties of the PG have not been reflected in the clinical or radiographic results.

J Clin Neurophysiol, 1988 Jul, 5(3), 211 - 35
Fluorescent voltage-sensitive dyes: applications for neurophysiology; Dasheiff RM; Voltage-sensitive dyes are a means to optically monitor changes in membrane potential . Their application in research has grown steadily over the last two decades as better dyes have been developed . The techniques presently in use are providing unique information about biologic systems from bacteria to the functional organization of primate occipital cortex . This review provides a history of the dyes, the data supporting their voltage sensitivity, and the techniques required for their use . The limitations in using and interpreting the voltage-sensitive dyes, as well as their diverse applications in all areas of research, especially neurophysiology, are comprehensively presented.

J Exp Med, 1988 Jul 1, 168(1), 267 - 77
Receptor analogs and monoclonal antibodies that inhibit adherence of Bordetella pertussis to human ciliated respiratory epithelial cells; Tuomanen E et al.; The adherence of Bordetella pertussis to human respiratory cilia is critical to the pathogenesis of whooping cough . To explore the development of agents that could interrupt adherence, the structure of the receptor on the ciliary surface was investigated . Using an in vitro adherence assay to human ciliated epithelial cells, galactose, lactose, and complex carbohydrates containing lactose eliminated adherence when preincubated with the bacteria . 10(-2) M galactose eluted adherent bacteria from cilia . B . pertussis and its two purified adhesins bound specifically to natural lactose-containing glycolipids in a TLC assay . mAbs to eukaryotic glycoconjugates with specificity for substituted galactose-glucose moieties blocked adherence when preincubated with ciliated cells . The carbohydrates that serve as receptors for B . pertussis on human cilia are galactose-glucose-containing glycolipids . Receptor analogs and anti-receptor antibodies effectively block adherence of B . pertussis to cilia and thus should be considered candidates for therapeutic intervention against disease.

Curr Eye Res, 1988 Jul, 7(7), 661 - 6
Rapid detection of herpes simplex virus (HSV) antigen in human ocular infections; Dunkel EC et al.; The new HERPCHEK (Dupont, No . Billerica, MA) enzyme immunosorbent assay (EIA) was used in a double-blind clinical study for rapid and specific detection of ocular herpes simplex virus (HSV) infection . This 4-hour assay can be used to demonstrate conclusively the presence of HSV antigen without culture and thereby rapidly differentiate between HSV and other clinically similar ocular infectious diseases . Ocular samples were collected from 180 individuals including 30 patients with acute HSV, 90 with latent HSV (ie, currently asymptomatic but with a positive history), 11 with acute or latent varicella zoster virus, 30 with nonherpetic infections (due to adenovirus, Acanthamoeba or bacteria), and 19 normal controls . A clinical diagnosis was made by one of us (DPL) and duplicate tear-film samples obtained by swabbing the conjunctival cul-de-sac and cornea . Coded samples were tested by routine viral culture on Vero cell monolayers and also were run independently in the HERPCHEK test . During active HSV infection, the HERPCHEK correlated 100% with clinical diagnosis, and virus culture correlated 90% with clinical diagnosis . In all latent HSV ocular infections, other nonherpetic ocular infections and normal samples, both the HERPCHEK and culture assays were negative.

Vopr Virusol, 1988 Jul-Aug, 33(4), 444 - 7
{Development of test systems of immunoenzyme analysis for the rapid diagnosis of rotavirus infection}; Bukrinskaia AG et al.; A test system of enzyme immunoassay (EIA) consisting of simian rotavirus SA-11 and rabbit antiserum has been developed for the detection of rotavirus antigen . Direct EIA was used for tests on stool specimens from 289 children varying in ages from 10 days to 12 years suffering from acute enteric infections and 56 normal children . The antigen was detected in 22.1% and 3.5%, respectively, in patients predominantly in the first 3 days of the disease and most frequently in cases running as gastroenteritis . The results are discussed with reference to the possibility of the development of rotavirus carrier state and mixed virus-bacteria infections.

Am J Obstet Gynecol, 1988 Jul, 159(1), 114 - 9
The value and limitations of the Gram stain examination in the diagnosis of intraamniotic infection; Romero R et al.; Gram stain examination of amniotic fluid is a method used for the rapid diagnosis of intraamniotic infection in patients with preterm premature rupture of membranes and preterm labor . The management of these patients relies heavily on the Gram stain results . Therefore, it is critical that the diagnostic value and limitations, optimal technique, and outcome correlates be precisely established . Most studies have focused on culture results rather than on Gram stain results to describe neonatal and maternal outcome . However, management is based on the Gram stain because culture results are not immediately available . One hundred eighty-seven amniocenteses were performed in 131 patients with preterm premature rupture of the membranes (n = 90) and preterm labor (n = 41) . Spun and unspun Gram stains were performed . Centrifugation of the sample did not improve the sensitivity of the technique significantly . The agreement between the two methods was substantial (kappa index 0.89, p less than 0.001) . The sensitivity of the Gram stain was 44.8% and the specificity was 97.6% . The sensitivity of the Gram stain was directly proportional to the number of bacteria present in amniotic fluid . In the presence of greater than 10(5) colony forming units per milliliter, 80% of the Gram stains were positive . The absence of both bacteria and white blood cells in a smear was associated with a negative culture of amniotic fluid in 95% of the cases . Clinical chorioamnionitis was associated with a positive Gram stain of amniotic fluid (p less than 0.001) . There was a trend toward a higher incidence of endometritis in patients with a positive Gram stain compared with those with a negative Gram stain (p = 0.07) . There was no neonatal infectious morbidity in patients with a true negative Gram stain . Patients with a false negative Gram stain had a 25% incidence of neonatal infectious complications (proved and suspected sepsis).

J Chromatogr, 1988 Jun 17, 442, 29 - 52
Conformational studies of bovine alkaline phosphatase in hydrophobic interaction and size-exclusion chromatography with linear diode array and low-angle laser light scattering detection; Krull IS et al.; Alkaline phosphatase has been studied in hydrophobic interaction chromatography (HIC), using a bonded C1-ether phase on a silica gel support, together with an aqueous salt gradient . Its behavior under various gradient elution conditions has demonstrated good chromatographic performance and retention of enzymatic activity under aqueous conditions . It has now been studied using linear photodiode array (LDA) spectroscopy in combination with low-angle laser light scattering (LALLS) in gradient elution HIC . HIC-LALLS permitted the use of routine salt gradients for collection of molecular weight information, despite small changes in the baseline, via computerized baseline subtraction . Size-exclusion chromatography (SEC)-LALLS measurements, under various isocratic conditions, meant to mimic HIC elution, have indicated the presence of monomer/dimer, dimer/trimer, or mainly trimer . aggregates of alkaline phosphatase can also be detected under salt gradient HIC conditions, but at lower levels relative to the monomer . This paper also describes the behavior of alkaline phosphatase when detected using LDA under various chromatographic, temperature, and concentration (injected) conditions . The results indicate a facile equilibrium of at least two monomeric forms of alkaline phosphatase of the same molecular weight, which change relative populations as a function of operational conditions . Most interesting is the suggestion that alkaline phosphatase undergoes rapid conformational interconversions on the chromatographic detection time scales, and that these interconverting conformations, concentration dependent, produce a novel dual wavelength ratioing, viz., a pseudo-Gaussian peak mimicking the chromatographic elution profile at either wavelength . The reasons for these observations and their possible use in future high-performance liquid chromatographic biopolymer studies are discussed and described.

J Theor Biol, 1988 Jun 7, 132(3), 307 - 18
Complex behaviour of the repressible operon; Sinha S et al.; The repressor-mediated repression process in bacteria is modelled using a gene-enzyme-endproduct control unit . A combined analytical-numerical study shows that the system, though stable normally, becomes unstable for super-repressing strains even at low values of the cooperativity of repression, provided demand for the endproduct saturates at large endproduct concentrations . In addition the system also shows bistability, i.e., the co-existence of a stable steady-state and a stable limit cycle . The tryptophan operon is used as a model system and the results are discussed in the light of differential regulation of gene expression in lower organisms, especially in mutant strains.

J Am Vet Med Assoc, 1988 Jun 1, 192(11), 1563 - 8
Tuberculosis in five basset hounds; Carpenter JL et al.; Five Basset Hounds (2 females and 3 males) under the age of 5 years, acquired systemic tuberculosis . We suspected tuberculosis in one dog, because it had histologic lesions similar to those in 4 dogs in which bacteria were identified as Mycobacterium avium complex . A review of canine tuberculosis revealed a similar diagnosis in a Basset Hound . The association of this infection in Basset Hounds suggests an inherited immunologic defect . Results of our survey suggest that the defect might exist in cell-mediated immunity.

Ann Surg, 1988 Jun, 207(6), 693 - 8
The role of feces, necrotic tissue, and various blocking agents in the prevention of adhesions; O'Leary JP et al.; Ischemic tissue and intraperitoneal bacteria have been ascribed an etiologic role in the production of intra-abdominal adhesions . To further elucidate the role of these stimuli and to evaluate the potential protective effect of various agents, peritonitis was induced in 160 Sprague-Dawley rats . The experiment was stratified into those animals with peritonitis plus necrotic tissue, solid feces, both, or neither . The agents tested were a nonsteroidal anti-inflammatory (ibuprofen), free radical scavenger (SOD), and an anticoagulant (heparin) . Death was less likely to occur in animals treated with heparin (3 of 40 vs . 12 of 40, p less than 0.01) or SOD (4 of 40 vs . 12 of 40, p less than 0.05) . Ibuprofen did not increase survival in this model . Heparin protected against adhesions in animals with an ischemic ileum of limb and without solid feces . In animals with a nonischemic isolated segment of ileum and solid feces, adhesion formation was increased in both the ibuprofen and the heparin treatment groups (p less than 0.05).

Ann Surg, 1988 Jun, 207(6), 686 - 92
The capacity of serum to support neutrophil phagocytosis is a vital host defense mechanism in severely injured patients; Polk HC Jr et al.; The opsonic capacity of patient serum was studied in 43 trauma patients of whom 13 recovered uneventfully, 21 developed major infection, and nine died, mostly of infection . Blood samples were taken within 24 hours of injury . Fifteen patients were studied serially of whom 14 developed severe infection and/or died . Opsonic capacity was determined by flow cytometry and measured as the ability of normal neutrophils to phagocytose killed bacteria previously incubated with patient serum . The most dilute sera reflected changes for better and worse most clearly . On initial assessment, those who died of sepsis showed a 61% mean fluorescent intensity (MFI), which was significantly lower than the 99% MFI for those who survived infection (p less than 0.01) and the 78% MFI of those who developed no infection (p less than 0.05) . Serial samples demonstrated a super serum response in four of seven patients surviving major sepsis but in none of the seven who died of infection.

Am J Med Sci, 1988 Jun, 295(6), 503 - 6
Effects of ribavirin on neutrophil function; Steele RW et al.; Ribavirin, a broad spectrum antiviral agent, has been shown to exhibit immunosuppressive activity . This property has raised concerns during clinical trials because candidates for antiviral therapy are those who may develop secondary bacterial infection . We therefore investigated a number of parameters of neutrophil function after the in vitro addition of various concentrations of ribavirin . At pharmacologic concentrations (1.25-7.5 micrograms/mL), percent killing of phagocytized bacteria was increased as measured by an acridine orange fluorochrome microassay, significantly higher at a concentration of 5 micrograms/mL (p less than .02) . There was no concomitant adverse effect on adherence, random migration, chemotaxis, opsonization or phagocytosis . The present data indicate that ribavirin has no acute suppressive effect on granulocyte function and may even enhance bacterial killing capabilities for treated patients.

J Clin Microbiol, 1988 Jun, 26(6), 1134 - 7
Isolation of Mobiluncus species from clinical specimens by using cold enrichment and selective media; Smith HJ et al.; New and selective Rlk and SA media, combined with cold enrichment at 4 to 5 degrees C, allowed isolation of Mobiluncus species from patients with bacterial vaginosis at higher rates than with conventional cultivation methods . Rlk medium consists of Columbia CNA agar supplemented with peptone, yeast extract, 5% laked rabbit or sheep blood, nalidixic acid, and tinidazole . SA medium consists of Columbia CNA agar supplemented with 2% rabbit serum, 1.6% laked rabbit or sheep blood, nalidixic acid, and tinidazole . Use of these selective media plus the cold enrichment technique permitted Mobiluncus species to propagate at rates similar to those of other anaerobic members of the vaginal flora.

Clin Cardiol, 1988 Jun, 11(6), 430 - 2
Aspergillus endocarditis in association with a false aortic aneurysm; Corrigan C et al.; A 32-year-old man was admitted with an intracerebral hematoma and subsequently with bilateral femoral emboli five months after aortic valve replacement . Blood cultures for bacteria and fungi were negative but microscopy of the embolus revealed Aspergillus fumigatus . At operation a large false aneurysm of the ascending aorta and vegetations on the prosthetic aortic valve were found . Aspergillus endocarditis is diagnosed antemortem in only 23% of fatal aortic aneurysms: blood cultures are positive in only 8% . Clinically a large vessel embolus is characteristic, being present in 83% and microscopy and culture of such an embolus if present are vital investigations.

J Clin Pathol, 1988 Jun, 41(6), 679 - 82
Comparison of conventional and single bottle system for blood cultures; Wilson JA et al.; A single bottle blood culture system (Oxoid Signal system) was compared with a conventional two bottle subculturing system . A total of 2016 routine blood samples yielded 186 (9%) clinically important isolates . Of these, 40 (21%) were isolated only in the Oxoid system and 30 (17%) only in the conventional system . One hundred and sixteen (62%) were isolated from both systems . Volume of blood was not significantly associated with the rate of detection but was significantly associated with the speed of detection . A continuation of the study with a Signal bottle and single conventional bottle confirmed initial results of the increased isolation rate using two systems . The Oxoid single bottle blood culture system was at least comparable with a conventional system in terms of rate of detection of organisms and was superior in terms of speed of detection.

J Clin Microbiol, 1988 Jun, 26(6), 1219 - 20
Modified Thayer-Martin medium for recovery of Nocardia species from contaminated specimens; Murray PR et al.; Specimens submitted for isolation of Nocardia species are frequently contaminated with other bacteria . Although decontamination with sodium hydroxide or benzalkonium chloride can remove these bacteria, this treatment is also toxic for Nocardia species . We demonstrate that modified Thayer-Martin medium can be used for selective isolation of Nocardia species.

Eur J Biochem, 1988 Jun 1, 174(2), 331 - 8
Kinetic and spectral studies on the redox forms of methanol dehydrogenase from Hyphomicrobium X; Frank J Jr et al.; Several reaction rate constants in the catalytic cycle of methanol dehydrogenase (EC 1.1.99.8) in vitro were determined with stopped-flow spectrophotometry . The studies revealed that the high pH required for adequate activity of the enzyme is related to the strong pH dependency of the oxidation rates of the reduced and semiquinone enzyme forms, MDHred and MDHsem, with the artificial electron acceptor Wurster's blue . The rate-limiting step in the catalytic cycle is associated with the conversion of oxidized enzyme-substrate complex (MDHox.S) into reduced enzyme (MDHred) and product . The effect of activator (ammonium salts) was also confined to this step, but even saturating concentrations were unable to remove the limitation completely . Making use of the large deuterium isotope effect associated with substrate oxidation in the absence of activator, the transient MDHox.C2H3OH complex could be isolated and its slow decomposition into MDHred and formaldehyde could be demonstrated . Further evidence is presented to support the view that the different absorption spectra observed originate from genuine redox forms of methanol dehydrogenase with different redox states of the cofactor PQQ and not from enzyme-electron acceptor complexes . The results confirm and extend our original view on the mechanism of action and contradict the mechanism proposed by others {Parkes, C . & Abeles, R . H . (1984) Biochemistry 23, 6355-6363}.

Radiology, 1988 Jun, 167(3), 669 - 71
Acute cholecystitis: diagnostic accuracy of percutaneous aspiration of the gallbladder; McGahan JP et al.; Results of gallbladder bile aspiration and culture were correlated with presence or absence of acute cholecystitis in 36 patients to test the role of these procedures in hospitalized patients with sepsis . Diagnostic aspiration of the gallbladder was performed in 11 patients, and in the remaining patients a combination of percutaneous aspiration, percutaneous cholecystostomy, or cholecystectomy was used . Bile culture was not helpful in the prediction of acute cholecystitis, since results were not available for a minimum of 24-48 hours after aspiration . In addition, gram-stained smears and bile cultures suffered from low sensitivity (48% and 38%, respectively); consequently, a negative test does not allow the diagnosis of acute cholecystitis to be excluded . Bile aspiration of the gallbladder thus has a limited role in the diagnosis of this condition.

J Immunol, 1988 Jun 1, 140(11), 3812 - 6
Mapping of biologically relevant sites on human IL-1 beta using monoclonal antibodies; Massone A et al.; mAb have been raised that recognize human IL-1 beta . Using overlapping peptide fragments expressed in yeast and bacteria, we have mapped the regions of the protein to which these antibodies bind . To assess the relevance of the different regions of IL-1 beta for the expression of its biologic activity, the ability of the antibodies to block IL-1 activity was assayed . Antibodies recognizing the regions 133-148 and 251-269 of human IL-1 beta could inhibit the activity of IL-1 beta, but not of IL-1 alpha, in two different biologic assays, the murine thymocyte proliferation and PGE2 release from human fibroblasts . Conversely, antibodies that recognize the region 218-243 have only a moderate inhibitory effect on the IL-1 beta biologic activity in both assays . Finally, an antibody mapping to the region 148-192 did not inhibit IL-1 beta activity either on thymocytes or on fibroblasts . It is suggested that IL-1 beta-induced cell activation involves different regions of the protein and that both N-terminal and C-terminal fragments are involved in the correct functioning of the IL-1 beta molecule.

Can J Microbiol, 1988 Jun, 34(6), 757 - 66
Ultrastructure of the in situ adherence of Mobiluncus to vaginal epithelial cells; De Boer JM et al.; From patients with bacterial vaginosis motile, anaerobic, comma-shaped bacteria can be isolated, which have recently been placed into the new genus Mobiluncus . In this study, electron microscopy was used to examine the in situ adherence of these motile curved rods to detached epithelial cells (comma cells) in vaginal fluid from two patients with bacterial vaginosis . Thin sections showed that the curved rods attached both directly to the epithelial cell surface and at various distances from it . It is concluded that after initial attachment these motile bacteria can grow at the epithelial cell surface in sessile microcolonies . Ruthenium red staining demonstrated a coating of precipitated glycocalyx material both on the surface of the curved rods and on their flagella . This may indicate that in situ the adherent curved rods were enclosed in a very hydrated matrix of exopolysaccharides . Conspicuous was the ability of the curved rods to attach to the epithelial cell surface via their cell tips . However, in situ no specialized bacteria cell surface structures were seen that might explain this polar attachment . Electron microscopy of pure cultures demonstrated that both Mobiluncus curtisii subsp . curtisii and Mobiluncus mulieris can produce a glycocalyx in vitro.

Arkh Anat Gistol Embriol, 1988 Jun, 94(6), 74 - 7
{Reaction of lymphocytes of the endometrium to the intrauterine administration of antigens}; Iakhnitsa AG et al.; In the mucosal membrane of the uterine horns in Wistar white rats mainly small and middle lymphocytes (predominantly T-cells) and rather small amount of plasmocytes and macrophages are presented . After intrauterine administration of bacteria and viruses (BCG vaccine, influenza inactivated liquid vaccine of A type) the local immune reaction develops according to the cellular type with maximum at the first week, before the total immune response, with its peak occurring on the 14th day of the experiment.

Anal Biochem, 1988 Jun, 171(2), 339 - 45
A general coupled spectrophotometric assay for decarboxylases; Burns DH et al.; A rapid, continuous spectrophotometric method has been developed for the assay of decarboxylases . The assay uses a coupled enzyme system in which liberated CO2 is reacted with phosphoenolpyruvate and phosphoenolpyruvate carboxylase to form oxaloacetate, which in turn is reduced by malate dehydrogenase to L-malate concomitantly with the oxidation of NADH to NAD . The resultant decrease in absorbance at 340 nm accurately reflects the activity of the decarboxylase . The method is capable of detecting the liberation of as little as 1 nmol of CO2/min and was tested in assays of lysine decarboxylase, orotidine-5'-phosphate decarboxylase, and 4'-phosphopantothenoyl-L-cysteine decarboxylase.

Proc Natl Acad Sci U S A, 1988 Jun, 85(12), 4262 - 6
No strict alignment is required between a transcriptional activator binding site and the "TATA box" of a yeast gene; Ruden DM et al.; GAL4 is a transcriptional activator of the galactose metabolism genes in the yeast Saccharomyces cerevisiae . We show that GAL4 expressed in yeast activated transcription equally well when a single GAL4 binding site was placed at any of nine positions upstream of the GAL1 (galactokinase gene) "TATA box." We chose a sufficient number of positions for the binding site to ensure that, in several of these positions, GAL4 was on the opposite side of the DNA helix with respect to the TATA box . Smaller GAL4 derivatives were similar to wild-type GAL4 in that they also activated transcription in a manner independent of the side of the DNA helix they bound with respect to the TATA box . Unlike wild-type GAL4, however, these smaller GAL4 derivatives activated transcription better when we placed a binding site progressively closer to the TATA box over a distance of 34 base pairs.

J Biochem (Tokyo), 1988 Jun, 103(6), 962 - 8
The protein responsible for center A/B in spinach photosystem I: isolation with iron-sulfur cluster(s) and complete sequence analysis; Oh-oka H et al.; The 9 kDa polypeptide from spinach photosystem I (PS I) complex was isolated with iron-sulfur cluster(s) by an n-butanol extraction procedure under anaerobic conditions . The polypeptide was soluble in a saline solution and contained non-heme irons and inorganic sulfides . The absorption spectrum of this iron-sulfur protein was very similar to those of bacterial-type ferredoxins . The amino acid sequence of the polypeptide was determined by using a combination of gas-phase sequencer and conventional procedures . It was composed of 80 amino acid residues giving a molecular weight of 8,894, excluding iron and sulfur atoms . The sequence showed the typical distribution of cysteine residues found in bacterial-type ferredoxins and was highly homologous (91% homology) to that deduced from the chloroplast gene, frxA, of liverwort, Marchantia polymorpha . The 9 kDa polypeptide is considered to be the iron-sulfur protein responsible for the electron transfer reaction in PS I from center X to {2Fe-2S} ferredoxin, namely a polypeptide with center(s) A and/or B in PS I complex . It is noteworthy that the 9 kDa polypeptide was rather hydrophilic and a little basic in terms of the primary structure . A three-dimensional structure was simulated on the basis of the tertiary structure of Peptococcus aerogenes {8Fe-8S} ferredoxin, and the portions in the molecule probably involved in contacting membranes or other polypeptides were indicated . The phylogenetic implications of the structure of the present polypeptide as compared with those of several bacterial-type ferredoxins are discussed.

Scanning Microsc, 1988 Jun, 2(2), 1123 - 38
The role of scanning electron microscopy in periodontal research; Carrassi A et al.; During recent years a great amount of research has led to a better understanding of the etiology, pathogenesis and pattern of progression of periodontal diseases . Scanning electron microscopy (SEM) has contributed to this improvement, mainly with respect to histology of periodontal tissues, the description of the morphology and distribution of bacteria on the exposed root surface, analysis of the host-parasite interactions on the gingival pocket wall, and morphological evaluation of root treatment . This review deals with all these topics . Unusual types of SEM research are also described and discussed . Uncommon sample preparation techniques for SEM in periodontal research are described . SEM in periodontal research should be of great application in the near future . Cathodoluminescence, back-scattered emission and immunolabelling techniques will be formidable tools in this field of dentistry.

Rev Rhum Mal Osteoartic, 1988 Jun, 55(8), 601 - 6
{Cellular targets of immunoglobulins eluted from the placenta}; Cohen JH et al.; Placenta eluted gammaglobulin were reported to have a beneficial effect among rheumatoid arthritis suffering patients . These gammaglobulin obtained by acidic elution from washed placentas (PEGG) were compared to plasmatic immunoglobulin (Ig) for their ability to inhibit mixed lymphocyte reaction (MLR), for their content of antibodies (Ab) directed against human tissues or bacteria and viruses . In addition, biochemical fractionations of PEGG were performed . Both PEGG and Ig from plasma (Sandoglobulin {SG}) contained anti nuclear Ab, Ab against smooth muscle, epithelium, vascular endothelium, synovial cells from rheumatoid arthritis synovium . Furthermore, PEGG at high concentration stained the Raji cell line that SG did not stain . PEGG inhibited undirectional MLR by acting on the stimulating cells . SG or plasmin digested Ig from retroplacental blood (Veinoglobulins, Merieux {VG}) also inhibited MLR but at 20 to 50 fold higher concentrations than PEGG . This activity was restricted to intact IgG, even further purified on protein A . Biochemical fractionation indicated that about 10% of PEGG consisted of free light chains, suggesting that deposits of free light chains might exist on placentas.

Agents Actions, 1988 Jun, 24(1-2), 114 - 9
Leukotriene B4 and inflammatory disease; McMillan RM et al.; Polymorphonuclear leukocytes (PMNL) are prominent at sites of acute inflammation . Their infiltration is stimulated under pathological conditions by a variety of agents which include bacteria, immune complexes and complement derived chemotactic peptides . Recently attention was focussed on the 5-lipoxygenase product leukotriene B4 (LTB4) which has been demonstrated to induce the key features associated with an acute inflammatory reaction . However, evidence supporting a pro-inflammatory role for LTB4, and therefore the anti-inflammatory efficacy of 5-lipoxygenase inhibitors, is largely circumstantial . Moreover, there are concerns that other chemotactic factors, notably C5a, may compensate for the absence of LTB4 . Here we challenge this view and, on the basis of recent experimental and clinical data suggest that LTB4 does not simply duplicate the activity of C5a . Instead we propose that their predominant site(s) of action differ in such a way that they may synergise in mediating PMNL recruitment.

J Virol, 1988 Jun, 62(6), 1948 - 55
Heterogeneity of adenovirus type 5 E1A proteins: multiple serine phosphorylations induce slow-migrating electrophoretic variants but do not affect E1A-induced transcriptional activation or transformation; Richter JD et al.; The 289-amino-acid product encoded by the adenovirus E1A 13S mRNA has several pleiotropic activities, including transcriptional activation, transcriptional repression, and when acting in concert with certain oncogene products, cell transformation . In all cell types in which E1A has been introduced (except bacteria), E1A protein is extensively posttranslationally modified to yield several isoelectric and molecular weight variants . The most striking variant is one that has a retarded mobility, by about Mr = 2,000, in sodium dodecyl sulfate gels . We have investigated the nature of this modification and have assessed its importance for E1A activity . Phosphorylation is responsible for the altered mobility of E1A, since acid phosphatase treatment eliminates the higher apparent molecular weight products . By using several E1A deletion mutants, we show that at least two seryl residues, residing between residues 86 and 120 and 224 and 289, are the sites of phosphorylation and that each phosphorylation can independently induce the mobility shift . However, E1A mutants lacking these seryl residues transcriptionally activate the adenovirus E3 and E2A promoters and transform baby rat kidney cells to near wild-type levels.

Arch Biochem Biophys, 1988 Jun, 263(2), 369 - 77
Sulfur-dependent inhibition of protein and RNA synthesis by iron-grown Thiobacillus ferrooxidans; Oliver DJ et al.; The addition of sulfur to iron-grown Thiobacillus ferrooxidans resulted in a rapid inhibition in the rates of protein synthesis and RNA synthesis . The inhibition of both functions was measured within 15 to 30 min and was maximal between 70 and 90% compared to the iron-grown controls . DNA synthesis, carbon dioxide fixation, and short-term ferrous oxidation rates of the bacteria growing on ferrous ions were not effected by sulfur addition, indicating that the sulfur addition was not perturbing general cellular energy metabolism . The inhibition caused by sulfur mimicked the effect of the RNA synthesis inhibitor, rifampicin, which inhibited both RNA and protein synthesis, but did not correspond with the translational inhibitor, chloramphenicol, which inhibited only protein synthesis in the first hour . Since chloramphenicol pretreatment did not block the sulfur effect, the inhibition of RNA synthesis following sulfur addition was not mediated through protein synthesis.

Gene, 1988 May 30, 65(2), 319 - 23
Entrapment of recombinant plasmids in SeaPlaque agarose plugs and their rapid purification from recircularised vectors; Upcroft P; A simple method is described which permits both the separation and concentration of circular recombinant plasmids from smaller plasmid vectors that are an undesirable by-product of a ligation reaction . SeaPlaque agarose plugs are used to entrap open-circular forms of recombinant plasmids during electrophoresis . In the example described over 98% of supercoiled, open-circular and linear forms of the 2.9-kb Bluescript plasmid vector, as well as the equivalent dimer forms, pass through the 1.4% SeaPlaque plug . Circular recombinant plasmids greater in length than the vector dimer are entrapped within the plug . By increasing the concentration of SeaPlaque, recombinants smaller than the vector dimer are retained in the trap, but with a concomitant increase in contamination by open-circular vector dimers . For most library constructions the high ratio of insert to vector used during the ligation reaction reduces the formation of vector dimers and makes this level of contamination inconsequential . The recombinant plasmids can be extracted readily from the SeaPlaque plug by excising it, melting the agarose and extracting with phenol . Alternatively, the excised plug can be melted and the recombinant plasmids used to transform bacteria, or mammalian cells, directly in the agarose . The procedure should be valuable for cloning large inserts for 'jumping' and 'linking' libraries, for large inserts in general where recircularisation is a low-frequency event, e.g., minichromosomes, for pulsed-field gel electrophoresis applications, and for hosts and vectors where genetic selection of the recombinant is not possible.

FEBS Lett, 1988 May 23, 232(2), 364 - 8
Photosynthetic reaction centre of Chloroflexus aurantiacus . Primary structure of M-subunit; Ovchinnikov YuA et al.; The M-subunit primary structure of the reaction centre (RC) from Chloroflexus aurantiacus composed of 306 amino acid residues has been determined by parallel analysis of the protein and corresponding DNA . The blocked N-terminus as well as replacement of the essential histidine liganding Mg of an accessory bacteriochlorophyll in purple bacteria by leucine distinguishes the M-subunit of Chloroflexus RC from that of purple bacteria.

Gene, 1988 May 15, 65(1), 83 - 91
Plasmid pIJ699, a multi-copy positive-selection vector for Streptomyces; Kieser T et al.; A plasmid vector, pIJ699, which provides positive selection for cloned DNA, was constructed using the replication functions of the Streptomyces wide-host-range multi-copy plasmid pIJ101 . The selection for inserts is based on the principle that plasmids with long uninterrupted perfect palindromes (inverted repeats) are 'not viable' in bacteria . For cloning, pIJ699 is digested with BglII . This produces two fragments, one of which is the linearized vector, with two arms of the palindrome at its ends, and the other is a 'spacer' which is needed to keep the inverted repeat sequences apart . The vector fragment is separated from the 'spacer' fragment and ligated with the DNA to be cloned . Plasmids with a fragment of cloned DNA, but not the circularized vector, give rise to thiostrepton-resistant transformants in Streptomyces lividans . The inverted repeat sequences contain a strong transcription terminator which reduces transcriptional read-through both in and out of the cloned fragment . This improves the stability of many hybrid plasmids and facilitates the study of the regulation of cloned genes.

FEBS Lett, 1988 May 9, 232(1), 221 - 6
The gamma-subunit of ATP synthase from spinach chloroplasts . Primary structure deduced from the cloned cDNA sequence; Miki J et al.; cDNA clones encoding the gamma-subunit of chloroplast ATP synthase were isolated from a spinach library using synthetic oligonucleotide probes . The predicted amino acid sequence indicated that the mature chloroplast gamma-subunit consists of 323 amino acid residues and is highly homologous (55% identical residues) with the sequence of the cyanobacterial subunit . The positions of the four cysteine residues were identified . The carboxyl-terminal region of the chloroplast gamma-subunit is highly homologous with those of the gamma-subunits from six other sources (bacteria and mitochondria) sequenced thus far.

Dtsch Med Wochenschr, 1988 May 6, 113(18), 711 - 8
{Mycobacterioses in patients with HIV infection}; Flepp M et al.; Mycobacterial infections were confirmed in 629 HIV-infected persons (1.4%) . Five patients had a generalized infection with Mycobacterium avium complex (MAC), four an extrapulmonary tuberculosis caused by Mycobacterium tuberculosis (M . tbc) . In general, the tuberculosis was the first severe opportunistic infection, while infections with MAC were more frequent in patients with already manifest AIDS . Common to all patients were a septic temperature and definite shift to the left of neutrophil granulocytes . Four of five patients with MAC also had diarrhoea, and three of four with tuberculosis additionally had peripheral lymphomas . The chest x-ray films were normal in six of the nine patients . The diagnosis was made in six patients primarily by the microscopic demonstration of acid-fast bacteria in lymph node tissue or stool, in three patients by culture from blood or liver tissue . Microscopic stool examination was helpful: in three of five patients with MAC and one of two with M . tbc in the stool culture, acid-fast bacteria had already been demonstrated . In an individual case MAC infections could not be distinguished either clinically or morphologically from infections with M . tbc, but only by culture.

J Appl Physiol, 1988 May, 64(5), 1953 - 9
Comparative physiology of rodent pulmonary macrophages: in vitro functional responses; Warheit DB et al.; Since toxicological testing of inhaled materials frequently requires utilization of several species, we have investigated pulmonary macrophage (PM) functional responses and compared the rat model with other rodents . Two strains of rats, three strains of mice, and one strain each of hamster and guinea pig were used in this study . The numbers of recovered cells by bronchoalveolar lavage generally correlated with animal body weight . The one exception was the Syrian Golden hamster from which increased numbers of macrophages were recovered . Cellular differential data obtained from lavaged cytocentrifuge preparations demonstrated that PM's account for greater than 97% of recoverable free lung cells for all species except the guinea pig, which contains a resident population of eosinophils . Cell morphology studies indicated that hamster PM exhibited the highest proportion of ruffled PM and demonstrated the highest phagocytic activity, whereas mouse PM phagocytic activity was significantly reduced compared with the other three species . In addition, chemotaxis studies showed that rat PM migrated best to zymosan-activated, complement-dependent chemoattractants, whereas hamster PM demonstrated an enhanced chemotactic response to N-formyl peptides . The results of these studies suggest that the rat may be the most efficient species for clearing inhaled particles, whereas hamsters and guinea pigs may best respond to bacteria.

Drug Intell Clin Pharm, 1988 May, 22(5), 399 - 401
Aseptic meningitis associated with naproxen; Sylvia LM et al.; A rare complication of nonsteroidal antiinflammatory drug (NSAID) use, particularly in patients with collagen vascular or autoimmune diseases, is aseptic meningitis . A healthy 21-year-old man receiving naproxen for muscle spasm was admitted with a chief complaint of severe headache . Approximately one week after beginning naproxen, the patient developed headache, fever (T 38.8 degrees C), shaking chills, and nuchal rigidity with occasional nausea and vomiting resulting in a 15-lb weight loss . Findings from a cerebrospinal fluid examination revealed polymorphonuclear pleocytosis and elevated protein, but no evidence of infection with bacteria, fungi, mycobacteria, or viral agents was noted . Within 36 hours of discontinuing naproxen, the meningitis-like symptoms markedly improved . Rechallenge with naproxen was not performed . In patients exhibiting meningitis-like symptoms, a thorough drug history, including that of recent or intermittent NSAID use, should be obtained.

J Steroid Biochem, 1988 May, 29(5), 505 - 10
Simultaneous quantification of five odorous steroids (16-androstenes) in the axillary hair of men; Nixon A et al.; Five 16-androstenes have been simultaneously quantified in extracts of the axillary hair of men (age range 18-40 years) using combined capillary gas chromatography-mass spectrometry, with specific ion monitoring . Quantities found (pmol/mg.hair, with approximate 24-h totals in parentheses) were: 5 alpha-androst-16-en-3-one, 0-15 (0-433); 4, 16-androstadien-3-one, 0-143 (0-4103); 5,16-androstadien-3 beta-ol, 0-3.5 (0-728); 5 alpha-androst-16-en-3 alpha-ol, 0-17 (0-1752) and 5 alpha-androst-16-en-3 beta-ol, 0-4 (0-416) . There were no significant relationships with age of the subjects for any of the steroids measured but significant relationships were found between the amounts of the two ketones and between 5 alpha-androst-16-en-3 alpha- and 3 beta-ols . These findings may indicate the existence of a pathway of metabolism in axillary bacteria in which 4,16-androstadien-3-one is reduced to 5 alpha-androst-16-en-3-one and thence to the 3 alpha- and 3 beta-alcohols . The data are discussed in the context of axillary odour because of the low olfactory thresholds of several of the 16-androstenes measured and because of the relatively large quantities found in some subjects.

J Pediatr Surg, 1988 May, 23(5), 476 - 7
Diffuse necrotizing tracheobronchitis: an acute and chronic disease; Rubin SZ et al.; Necrotizing tracheobronchitis (NTB) is characterized by acute episodes of airway obstruction, hypercarbia, and lack of chest movement in mechanically ventilated neonates . Emergency bronchoscopic removal of necrotic tissue is essential for survival . Although postmortem lesions extend into smaller bronchi, survivors have not demonstrated residual tracheobronchial abnormalities . Two infants were treated successfully for NTB but succumbed to diffuse tracheobronchial strictures with progressive pulmonary hyperinflation . A third neonate with esophageal atresia and left pulmonary agenesis developed NTB . Despite initial postbronchoscopic improvement, the infant died at age 6 weeks with diffuse obstructing NTB . All three infants required endotracheal intubation and mechanical ventilation . High-frequency jet ventilation was not used . Tracheal cultures for fungi, bacteria and viruses were negative . Successful treatment of NTB may be followed acutely by recurrence of NTB and chronically by diffuse tracheobronchial strictures and emphysema.

J Pathol, 1988 May, 155(1), 35 - 9
Fatal Legionnaires' pneumonia: frequency of legionellosis in autopsied patients with pneumonia from 1969 to 1985; Schurmann D et al.; In 89 of 3027 (2.9 per cent) patients in whom pneumonia was found at autopsy during the period from 1969 to 1985, legionella pneumonia was diagnosed . Legionellae were demonstrated through the direct fluorescent antibody test by screening lung tissue sections . All 89 cases were caused by Legionella pneumophila . Suspicious typical rod-shaped organisms were demonstrated in all cases by the modified Dieterle stain whereas conventional histological stains failed to detect causative bacteria . The frequency per year of legionellosis in patients with pneumonia at autopsy ranged from 1.4 per cent (3/212 cases) to 6.8 per cent (12/177 cases) . In patients who have died of unexplained pneumonia, examinations for the demonstration of legionellae should be carried out.

Fundam Appl Toxicol, 1988 May, 10(4), 571 - 89
Genotoxicity of complex mixtures: strategies for the identification and comparative assessment of airborne mutagens and carcinogens from combustion sources; Lewtas J; Two strategies for assessment of the toxicity of complex mixtures are described and illustrated with examples from genotoxicity studies of complex combustion mixtures . The first, a strategy for identifying biologically active compounds or compound classes in complex mixtures, is called bioassay-directed fractionation and characterization . The identification and assessment of mutagens and potential carcinogens in complex mixtures has been significantly advanced by the use of short-term genetic bioassays . Bioassay-directed fractionation coupled with new analytical characterization methods has provided the tools needed to more efficiently identify potential carcinogens in complex combustion emissions and urban air samples . These studies have shown that a significant portion of the mutagenicity in combustion emissions and urban air is found in fractions more polar than polynuclear aromatic hydrocarbons (PAHs) . A second strategy, the comparative potency method, provides an approach to evaluating the relative toxicities of a series of mixtures . The comparative mutagenicity and carcinogenicity of a series of combustion emissions has been assessed using dose-response studies in bacteria, mammalian cells, and rodents . Comparative mutagenic and tumorigenic emission rates or emission factors provide a means to directly compare the relative hazard of the sources . This data base has also been used to develop a comparative risk assessment methodology for combustion emissions.

J Am Dent Assoc, 1988 May, 116(6), 700 - 4
The effect of sealants on dental caries: a review; Swift EJ Jr; Many dentists are concerned that caries may progress if inadvertently sealed within a tooth by a pit and fissure sealant . Several research studies have been performed to determine whether this is an important concern . This report reviews the methods and results of those studies.

J Allergy Clin Immunol, 1988 May, 81(5 Pt 2), 1004 - 9
Recent advances in immunologic reactivity in otitis media with effusion; Bernstein JM; The role of IgE-mediated hypersensitivity in the development of middle ear disease has not been completely resolved . However, on the basis of our investigations and those of other laboratories, we suggest that approximately two thirds of patients with chronic recurrent otitis media do not have allergies . The other third may have allergic rhinitis, and this allergic rhinitis could play a direct role in producing eustachian tube dysfunction in recurrent otitis media . However, viral infections of the upper respiratory tract may also induce IgE-mediated release of mast cell inflammatory mediators, and could cause otitis media on the basis of viral infection alone . Subtle immunologic deficiencies involving the IgG2 subclass and other immunoglobulin subclasses may also be lower in otitis-prone children, and this may be a genetically inherited disorder . Finally, the possibility of food allergy in otitis media must be considered, particularly in the young otitis-prone child with chronic recurrent otitis media.

Med Clin North Am, 1988 May, 72(3), 613 - 21
Metronidazole; Scully BE; Metronidazole is a highly effective therapy for anaerobic infections and a variety of protozoal and parasitic diseases . Its pharmacokinetics, toxicities, and unique mode of action are reviewed in detail . Indications for use and respective dosages are suggested.

Exp Hematol, 1988 May, 16(4), 274 - 80
The dimer of hemoregulatory peptide (HP5B) stimulates mouse and human myelopoiesis in vitro; Laerum OD et al.; A synthetic analogue of a pentapeptide associated with mature granulocytes has been described earlier and shown to suppress myelopoietic colony formation in vitro in concentrations from 10(-13) to 10(-6) M . By oxidation of the peptide, a dimer will rapidly occur by formation of disulfide bridges between cysteine residues . We here demonstrate that this dimer has the opposite effects of the monomer . For both mouse and human granulocyte-macrophage colony-forming units (CFU-GM), a dose-dependent enhancement of colony formation was observed in the dose range 10(-16) to 10(-5) M, where a saturation level was reached above 10(-8) M . At low doses of colony-stimulating activity (CSA) and in the linear stimulating phase, an up to ten times increase of colony formation was seen, whereas at higher doses the effect was less pronounced . Also at the plateau level of CSA stimulation an increased colony yield was seen . All types of colonies were stimulated . The dimer itself had no colony-stimulating factor activity and was not toxic to bone marrow cells in suspension cultures up to 24 h . Upon reduction of the dimer by use of sulfhydryl compounds, inhibitory effects on CFU-GM were restored . The peptide had no effect on the phagocytic process in human granulocytes, including attachment and internalization of bacteria or Zymosan particles . The monomerdimer equilibrium of hemoregulatory peptide may constitute a new mechanism for proliferative regulation of myelopoietic cells.

Clin Neuropathol, 1988 May-Jun, 7(3), 120 - 30
Sural nerve biopsy findings in leprosy: a qualitative and quantitative light and electron microscope study in 4 treated cases of the lepromatous spectrum; Gibbels E et al.; Reports on biopsy findings in multifascicular nerves in lepromatous leprosy (LL) are rare and detailed morphometrical data are not available . In a case of early LL with normal electrodiagnostic findings in sural nerve, the present study revealed marked segmental de- and remyelination concomitant with the sequelae of considerable Wallerian degeneration of preferentially small myelinated fibers (MF) in spite of a normal number/nerve and density/mm2 . Segmental de- and remyelination of several consecutive internodes in teased fibers suggests continuous bacterial spread via Schwann cells . In 2 more advanced LL-cases, nervous parenchyme was severely reduced, in a borderline lepromatous (BL) case obviously in part caused by cell infiltrates and granulomata . Distinct fascicle differences in MF-involvement were demonstrated by coefficients of variation of MF/mm2 and teased fiber preparations in LL, consistent with the hypothesis of initial focal spread of bacteria . Numbers and densities of endoneurial vessels were increased only in the later stages of LL . Enlargement of endoneurial area, due to different factors, was encountered except for the most severe LL-case with extensive endoneurial collagenization . Morphometric results were compared with those of other neuropathies . Intact and degenerating bacteria mostly in phagosomes of Schwann cells associated with unmyelinated axons and in macrophages were seen only in the early LL- and the BL-case . Sparse mononuclear cell infiltrates and small focal necrosis, present even in LL-cases, underline the complex pathogenesis of nerve fiber involvement.

Vet Microbiol, 1988 May, 17(1), 65 - 74
Mechanism of protection induced in mice against Erysipelothrix rhusiopathiae infection by treatment with porcine antiserum to the culture filtrate of an attenuated strain; Sawada T et al.; The mechanism of protection induced in mice against challenge with a virulent strain of Erysipelothrix rhusiopathiae by porcine antiserum to the culture filtrate (CF) of an attenuated strain was investigated . Death and bacterial growth in the spleens of mice challenged with the virulent strain were completely prevented by treatment with the antiserum . The protective effect of the serum was markedly decreased in mice in which polymorphonuclear leucocytes (PMN) were depleted by cyclophosphamide (CY) treatment but not in mice in which macrophages were blocked selectively by carrageenan (CG) . The phagocytic rate of PMN and the number of bacteria ingested by PMN were significantly higher in mice treated with the antiserum than in mice treated with normal serum . These results indicate that anti-CF serum exerts its protective effect by opsonic activity and that opsonized E . rhusiopathiae are eliminated mainly by PMN.

Stain Technol, 1988 May, 63(3), 155 - 8
A quick preparative method for electron microscopy observations of delicate objects using alginate embedding medium; Tamponnet C et al.; A quick, safe method has been devised for embedding small or fragile specimens and keeping delicate structures intact . Cells or organisms to be embedded are placed in a viscous sodium alginate solution (1-2%), which is then polymerized in 100 mM calcium chloride . The resulting gel is easily dehydrated, embedded in resin and sectioned for electron microscopy . This method, the alginate gel portion of which was originally developed for the immobilization of Euglena, allows direct observation of each element of the specimens in micrographs . If desired, the alginate can be removed after sectioning by sequestration of calcium in a 20 mM solution of sodium citrate or a 10 mM solution of EGTA . Cells and organelles in the sections respond normally to standard staining procedures.

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