Genitourinary issues during spaceflight: a review.

The genitourinary (GU) system is not uncommonly affected during previous spaceflights. GU issues that have been observed during spaceflight include urinary calculi, infections, retention, waste management, and reproductive. In-flight countermeasures for each of these issues are being developed to reduce the likelihood of adverse sequelae, due to GU issues during exploration-class spaceflight, to begin in 2018 with flights back to the Moon and on to Mars, according to the February 2004 Presendent's Vision for US Space Exploration. With implementation of a robust countermeasures program, GU issues should not have a significant threat for mission impact during future spaceflights.

Inhibition of nanobacteria by antimicrobial drugs as measured by a modified microdilution method.

Compounds from 16 classes of antimicrobial drugs were tested for their abilities to inhibit the in vitro multiplication of nanobacteria (NB), a newly discovered infectious agent found in human kidney stones and kidney cyst fluids from patients with polycystic kidney disease (PKD). Because NB form surface calcifications at physiologic levels of calcium and phosphate, they have been hypothesized to mediate the formation of tissue calcifications. We describe a modified microdilution inhibitory test that accommodates the unique growth conditions and long multiplication times of NB. This modified microdilution method included inoculation of 96-well plates and determination of inhibition by periodic measurement of the absorbance for 14 days in cell culture medium under cell culture conditions. Bactericidal or bacteriostatic drug effects were distinguished by subsequent subculture in drug-free media and monitoring for increasing absorbance. NB isolated from fetal bovine serum (FBS) were inhibited by tetracycline HCl, nitrofurantoin, trimethoprim, trimethoprim-sulfamethoxazole, and ampicillin at levels achievable in serum and urine; all drugs except ampicillin were cidal. Tetracycline also inhibited multiplication of isolates of NB from human kidney stones and kidney cyst fluids from patients with PKD. The other antibiotics tested against FBS-derived NB either had no effect or exhibited an inhibitory concentration above clinically achievable levels; the aminoglycosides and vancomycin were bacteriostatic. Antibiotic-induced morphological changes to NB were observed by electron microscopy. Bisphosphonates, aminocaproic acid, potassium citrate-citric acid solutions, and 5-fluorouracil also inhibited the multiplication of NB in a cidal manner. Insights into the nature of NB, the action(s) of these drugs, and the role of NB in calcifying diseases may be gained by exploiting this in vitro inhibition test system.

[Culture and identification of nanobacteria in bile].

OBJECTIVE: To study the distribution and identification of nanobacteria in bile and to evaluate the identifying methods of nanobacteria. METHODS: RPMI1640 culture or RPMI1640 culture with 10% heat-inactivated gamma-FBS was added into 75 samples of cystic bile from gallbladders resected in operation. Nanobacteria were identified by immunohistochemical staining, transmission electron microscopy (TEM), and calcific staining. RESULTS: Nanobacteria were found in 45 bile samples with a positive rate of 61.3%. The positive rate of nanobacteria was 75.7% among 37 bile samples with white precipitate adhering to the tube, and was 47.4% among the samples with flocculent precipitate or without precipitate (P < 0.05). The immunohistochemically confirmed presence of nanobacteria was re-confirmed by TEM in all the positive samples. The positive rate, sensitivity, specificity, false positive rate and false negative rate of calcific staining were 38.7%, 58.7%, 93.1%, 6.9% and 41.3% respectively. CONCLUSION: Immunohistochemistry with monoclonal antibody of nanobacteria associated with TEM is useful in identifying nanobacteria. Calcific staining is of great value to identification of nanobacteria. Precipitation of white floccules adhering to the tube is an important microbiological characteristic of nanobacteria.

Nanobacteria: controversial pathogens in nephrolithiasis and polycystic kidney disease.

Nanobacteria are unconventional agents 100-fold smaller than common bacteria that can replicate apatite-forming units. Nanobacteria are powerful mediators of biogenic apatite nucleation (crystal form of calcium phosphate) and crystal growth under conditions simulating blood and urine. Apatite is found in the central nidus of most kidney stones and in mineral plaques (Randall's plaques) in renal papilla. The direct injection of nanobacteria into rat kidneys resulted in stone formation in the nanobacteria-injected kidney during one month follow-up, but not in the control kidney injected with vehicle. After intravenous administration in rats and rabbits, nanobacteria are rapidly excreted from the blood into the urine, as a major elimination route, and damage renal collecting tubuli. Nanobacteria are cytotoxic to fibroblasts in vitro. Human kidney cyst fluids contain nanobacteria. Nanobacteria thus appear to be potential provocateurs and initiators of kidney stones, tubular damage, and kidney cyst formation. It is hypothesized that nanobacteria are the initial nidi on which kidney stone is built up, at a rate dependent on the supersaturation status of the urine. Those individuals having both nanobacteria and diminished defences against stone formation (i.e. genetic factors, diet and drinking habits) could be at high risk. Kidney cyst formation is hypothesized to involve nanobacteria-induced tubular damage and defective tissue regeneration yielding cyst formation, the extent of which is dependent on genetic vulnerability.

[Nanobacteria. An experimental neo-lithogenesis model]. / Nanobacteria. Un modelo de neo-litogénesis experimental.

OBJECTIVE: To study the phenomenon of biomineralization in an experimental model of lithogenesis by percutaneous renal punction without laparotomy and no antibiotics. METHODS/RESULTS: The study comprised 4 Brown-Norway rats. Nanobacteria inoculum (X, 2X and 4X) was administered to three rats and the remaining one was used as control. The analytical and radiological findings showing the development of obstructive pyelocaliceal lithiasis in the kidneys of rats no. 2 and 3 are presented. CONCLUSIONS: Translumbar percutaneous renal puncture has permitted performing laparotoy without antibiotic coverage, which was the main difficulty of the experimental model of lithogenesis. Nanobacteria were cultured successfully, but not without difficulty, and formation of calculi in the rat pyelocaliceal system was achieved. This experimental model will provide further insight into lithogenesis and will allow us to find the answers to some of the many questions concerning this condition that remain.

Endotoxin and nanobacteria in polycystic kidney disease.

BACKGROUND: Microbes have been suspected as provocateurs of polycystic kidney disease (PKD), but attempts to isolate viable organisms have failed. Bacterial endotoxin is the most often reported microbial product found in PKD fluids. We assessed potential microbial origins of endotoxin in cyst fluids from 13 PKD patients and urines of PKD and control individuals. METHODS: Fluids were probed for endotoxin and nanobacteria, a new bacterium, by the differential Limulus Amebocyte Lysate assay (dLAL), genus-specific antilipopolysaccharide (LPS) antibodies, monoclonal antibodies to nanobacteria, and hyperimmune serum to Bartonella henselae (HS-Bh). Selected specimens were also assessed by transmission electron microscopy (TEM) and nanobacterial culture methods. RESULTS: LPS or its antigenic metabolites were found in more than 75% of cyst fluids tested. Nanobacteria were cultured from 11 of 13 PKD kidneys, visualized in 8 of 8 kidneys by TEM, and immunodetected in all 13 PKD kidneys. By immunodetection, nanobacterial antigens were found in urine from 7 of 7 PKD males, 1 of 7 PKD females, 3 of 10 normal males, and 1 of 10 normal females. "Nanobacterium sanguineum" was dLAL positive and cross-reactive with antichlamydial LPS and HS-Bh. Some cyst fluids were also positive for LPS antigens from Escherichia coli, Bacteroides fragilis and/or Chlamydia, and HS-Bh, as were liver cyst fluids from one patient. Tetracycline and citrate inhibited nanobacterial growth in vitro. CONCLUSION: Nanobacteria or its antigens were present in PKD kidney, liver, and urine. The identification of candidate microbial pathogens is the first step in ascertaining their contribution, if any, to human disease.

Nanobacteria: an infectious cause for kidney stone formation.

BACKGROUND: Nanobacteria are cytotoxic, sterile-filterable, gram-negative, atypical bacteria detected in bovine and human blood. Nanobacteria produce carbonate apatite on their cell walls. Data on Randall's plaques suggest that apatite may initiate kidney stone formation. We assessed nanobacteria in 72 consecutively collected kidney stones from Finnish patients. METHODS: Nanobacteria and kidney stone units were compared using scanning electron microscopy (SEM). Demineralized kidney stones were screened for nanobacteria using a double-staining method and a specific culture method. Isolated nanobacteria were analyzed for mineral formation in vitro with Ca and 85Sr incorporation tests. RESULTS: SEM highlighted the resemblance in size and morphology of nanobacteria and the smallest apatite units in the kidney stones. Nanobacterial antigens could be detected after the demineralization of the stones in 1 N HCl. Nanobacteria were surprisingly resistant to this treatment, and cultures could be established from 93.1% of the stones. Only struvite stones had common bacteria, in addition to the nanobacteria. When the results of all of the assays were combined, 70 of the 72 stones (that is, 97.2%) were nanobacteria positive. Although apatite stones indicated highest nanobacteria antigen signals, the overall nanobacteria positivity did not depend on the stone type. The isolated nanobacteria produced apatite stones in vitro, measured by Ca and 85Sr incorporation. CONCLUSIONS: We propose that kidney stone formation is a nanobacterial disease analogous to Helicobacter pylori infection and peptic ulcer disease. Both diseases are initiated by bacterial infection and subsequently endogenous and dietary factors influence their progression.

Nanobacteria: an alternative mechanism for pathogenic intra- and extracellular calcification and stone formation.

Calcium phosphate is deposited in many diseases, but formation mechanisms remain speculative. Nanobacteria are the smallest cell-walled bacteria, only recently discovered in human and cow blood and commercial cell culture serum. In this study, we identified with energy-dispersive x-ray microanalysis and chemical analysis that all growth phases of nanobacteria produce biogenic apatite on their cell envelope. Fourier transform IR spectroscopy revealed the mineral as carbonate apatite. The biomineralization in cell culture media resulted in biofilms and mineral aggregates closely resembling those found in tissue calcification and kidney stones. In nanobacteria-infected fibroblasts, electron microscopy revealed intra- and extracellular acicular crystal deposits, stainable with von Kossa staining and resembling calcospherules found in pathological calcification. Previous models for stone formation have led to an hypothesis that elevated pH due to urease and/or alkaline phosphatase activity is a lithogenic factor. Our results indicate that carbonate apatite can be formed without these factors at pH 7.4, at physiological phosphate and calcium concentrations. Nanobacteria can produce apatite in media mimicking tissue fluids and glomerular filtrate and provide a unique model for in vitro studies on calcification.

Slime production by coagulase-negative staphylococci isolated in chronic blepharitis.

BACKGROUND: The purpose of the study was to determine the impact of slime-producing strains of coagulase-negative staphylococci (CNS) on non-ulcerative blepharitis. Formerly considered harmless organisms, CNS are now recognised as opportunistic pathogens. Although these microorganisms are a component of normal conjunctival flora, they often produce the typical signs and symptoms of chronic staphylococcal blepharoconjunctivitis. Certain strains produce a polysaccharide extracellular material called "slime". Slime production is considered to be associated with the virulence of the organism. METHODS: Swabs were taken from the lids of 38 eyes of 19 patients with chronic non-ulcerative blepharitis and cultured for CNS. A group of 42 normal control eyes were similarly sampled. The strains of CNS isolated from 26 eyes (68.4%) of the patients with blepharitis and 25 eyes (59.5%) of the normal subjects were studied for slime layer production. RESULTS: No significant difference was found between normal subjects and patients in the incidence of slime producing CNS strains from the conjunctiva. The antibiotic sensitivity profiles of the slime-producing strains were no different from the slime-negative isolates in the blepharitis (p=0.85) and normal group (p=0.25). CONCLUSIONS: Our data suggest that slime production by CNS does not play a significant role in the pathogenesis of staphylococcal blepharitis.

A case of human orotracheal myiasis caused by Wohlfahrtia magnifica.

A case of orotracheal myiasis in an 80-year-old man in coma for 1 week in the intensive care unit of Baskent University Hospital, Ankara, Turkey, is reported. A number of larvae of Wohlfahrtia magnifica were recovered repeatedly from the mouth and from the intubation tube of the patient and were reared to adults to facilitate identification. This case history points out the risky of myiasis caused by W. magnifica in the unconscious, debilitated patient.

[Prevalence of G. vaginalis, Mycoplasma, Ureaplasma, T. vaginalis, yeast, N. gonorrhoeae and other bacteria in women with vaginal discharge]. / Vajinal akintisi olan kadinlarda G. vaginalis mikoplazma, üreaplazma, T vaginalis, maya ve N. gonorrhoeae ve diger bakterilerin sikligi.

Vaginal discharge of 118 women attended to outdoor clinics of obstetrics and gynaecology at Medical Faculty of Ankara University (A.U.T.F.), have been examined. Direct microscopy has been made by means of wet mount, Gram and Giemsa staining. Vaginal pH has been measured, by 10% KOH the presence of "fishy odor" has been investigated. For culturing vaginal secretions in Amies transport medium have been brought to Microbiology Department of Ankara University, Faculty of Medicine Inoculations have been made on to Blood agar, MacConkey agar, Sabouraud Dextrose agar, New York City. Medium, % 5 Human blood bilayer Tween 80 Medium (HBT), Vivey Trichomonas Agar Medium and into M-broth and U9 broth. In this study, we have isolated 9 (7.62%) Gardnerella vaginalis, 41 (33.89%) Ureaplasma, 13 (11.01%) Mycoplasma, 25 (21.18%) yeasts, 4 (3.38%) Trichomonas vaginalis and 43 (36.46%) various bacteria, 26 cases were found to have polymicrobial etiology.

[Antibiotic susceptibility of group A beta hemolytic Streptococcus]. / A grubu beta hemolitik streptokoklarin antibiyotiklere duyarliligi.

In this study, we researched the susceptibility of group A Beta-hemolytic Streptococcus to antibiotics which are isolated from 218 Aydinlikevler primary blind boarding school students aged between 7-16 and a group of patient's throat and nose culture with acute and chronic upper respiratory tract infection. The susceptibility to 9 antibiotics of (100) group A Beta hemolytic Streptococcus which were isolated 52 from throat, 48 from nasal discharge were examined by the method of disc diffusion test after determination of the colony, hemolysis and staining characteristics also the sensibility to optochine and bacitracin (0.04 U) discs colonies. The antibiotic resistance ratio are observed as 24%. For Penicillin-G, 59% for Ampicillin, 17% for Amoxicillin, 31% for Methicillin, 22% for Cephalexin, 33% for Carbenicillin, 77% for Lincomycin, 60% for Trimethoprim/Sulphamethoxazole, 26% for Chloramphenicol. This finding shows that group A Beta hemolytic Streptococcus are getting being resistant to this 9 antibiotics. Also this results indicate that the treatment after antibiotic susceptibility test of group A Beta hemolytic Streptococcus is more effective.

[Immunoelectrophoretic analysis of Escherichia coli strains (EPEC, ETEC, EIV and UPEC) (1)]. / Escherichia coli (EPEC-ETEC-EIV ve UPEC) Suslarinin immünoelektroforetik analiz örnekleri (1).

Water extracts of Escherichia coli "O" and "K" antigen test strains (EPEC, ETEC, EIEC, UPEC) were examined in immunodiffusion and immunoelectrophoresis tests. The precipitation arcs corresponding to the O-antigen specificity and to the thermostable polysaccharide K antigen were easy to identify. All strains gave an O antigen precipitation arc found either on the anodic or the cathodic side of application basin and close to this. The so-called enteropathogenic types (from infantile diarrhoea) had a cathodic O antigen arc type; from dysentery-like disease had a negatively charged O-antigen, but no special thermostable K-antigen. Thus E. coli strains which may invade the tissues when conditions allow have a negatively charged surface antigen, either O-antigen lipopolysaccharide or both. Acidic components were found in the anodic O-antigen.