Dysbiotic microbiota in autistic children and their mothers: persistence of fungal and bacterial wall-deficient L-form variants in blood.

Based on our hypothesis for existing microbiota of wall-deficient variants (L-forms) in human blood, we created an innovative methodology, which allowed for the development of L-form populations from blood of all investigated people. In contrast to healthy controls, blood L-forms from autistic children and their mothers converted under appropriate conditions of cultivation into detectable opportunistic bacteria and fungi, а process demonstrated by light and transmission electron microscopy. It can be distinguished into two types of states - "eubiotic" blood microbiota in healthy individuals, and "dysbiotic" in autistic children and their mothers. Remarkably, the unifying finding for autistic children and their mothers was the presence in blood of wall-free variants from life-cycle of filamentous fungi. Increased specific IgG, IgM and IgA, together with typical mold growth were a decisive argument for proven presence of Aspergillus fumigatus in almost all of the autistic children. As it was demonstrated in our previous study, filterable L-forms can be transmitted by vertical pathway from mother to child before birth. Thus, it can be suggested that autistic children may be born already colonized with fungi, while a "silent aspergillosis" could contribute or even be a leading cause for neurodevelopmental disorders in the early childhood.

Psycho-emotional status but not cognition is changed under the combined effect of ionizing radiations at doses related to deep space missions.

Human spaceflight launch is the big challenge that the humanity work on. The astronauts' task performance vulnerability to ionizing radiations is one of the major factors limiting deep space missions. In this work, we study the effect of ionizing radiations (γ-quanta and 12C6+ in combination) on cognitive abilities and psycho-emotional status of Wistar rats. Irradiation led to the hyperlocomotion, increase of anxiety-like behavior, suppression of depressive-like behavior and enhancement of spatial learning. These data are consistent with the neurochemical/molecular analysis: enhanced monoaminergic innervation within the hypothalamus (HYP), inhibition of serotonin turnover in the prefrontal cortex and neurokenin 1 receptor overexpression in the amygdala (AMY). In addition, we observe decreased expression of certain biomolecules in the AMY (5-HT2c and 5-HT3) and in the HYP (5-HT2a, 5-HT4 and VMAT2) that can be explained as neuroadaptive changes. Thus, the ionizing radiation exposure significantly modulates the psycho-emotional status. With that, for the first time we received data that radiation effects in the doses and composition of interplanetary space (in terrestrial modeling) could be relatively safe for cognitive functions.

An excess of massive stars in the local 30 Doradus starburst.

The 30 Doradus star-forming region in the Large Magellanic Cloud is a nearby analog of large star-formation events in the distant universe. We determined the recent formation history and the initial mass function (IMF) of massive stars in 30 Doradus on the basis of spectroscopic observations of 247 stars more massive than 15 solar masses ([Formula: see text]). The main episode of massive star formation began about 8 million years (My) ago, and the star-formation rate seems to have declined in the last 1 My. The IMF is densely sampled up to 200 [Formula: see text] and contains 32 ± 12% more stars above 30 [Formula: see text] than predicted by a standard Salpeter IMF. In the mass range of 15 to 200 [Formula: see text], the IMF power-law exponent is [Formula: see text], shallower than the Salpeter value of 2.35.

Mother-to-newborn transmission of mycobacterial L-forms and Vδ2 T-cell response in placentobiome of BCG-vaccinated pregnant women.

The ability of bacteria to exist as a population of self-replicating forms with defective or entirely missing cell wall (L-forms) is an adaptive mechanism for their survival and reproduction under unfavorable conditions. Bacterial mother-to-fetus transfer is a universal phenomenon in the animal kingdom. However, data about vertical transfer of L bacterial forms are extremely scarce. Bacille Calmette-Guérin is an attenuated strain of M. bovis and the only licensed vaccine used for tuberculosis prevention. We already have shown that filterable L-forms of BCG exist freely in the vaccine and are able to reproduce and to form colonies. The present study was focused on the placental microbiome in the context of mother's BCG vaccination. Here we report an isolation of filterable mycobacterial L-form cultures from gestational tissues and blood of healthy newborns delivered by healthy BCG-vaccinated mothers after normal pregnancy. Of note, vertically transmitted mycobacterial L-forms as a part of placentobiome of the pregnant women didn't influence the number of resident pathogen-reactive Vδ2 cells. Placenta colonization with mycobacterial L-forms occurs by maternal blood-to-decidua transfer very early in gestation. Together, these data showed that BCG L-forms have the capacity to pass trans-placental barrier and that maternal BCG vaccination affects the placentobiome.

[The antiviral activity of the adamantane derivatives against the influenza virus A (H1N1) pdm2009 model in vivo].

For the first time in vivo, the model of the viral pneumonia in mice was used to study the antiviral activity against influenza A virus (H1N1) pdm09 synthetic derivatives of adamantane series including the amino acid residues and lipoid acid. It was found that the adamantane derivatives with histidine, serine, and lipoid acid could inhibit the rimantadine-resistant strain of the influenza A (H1N1) pdm09. As a result, the lifespan of the mice infected with the virus has increased by 1.6 times with respect to viral control. Thus, the possibility of restoration of antiviral properties of rimantadine both in vitro and in vivo by introducing into its molecular structure new functionally active groups was tested.

Antibacterial and antimycotic activity of a cross-linked electrospun poly(vinyl pyrrolidone)-iodine complex and a poly(ethylene oxide)/poly(vinyl pyrrolidone)-iodine complex.

Photo-cross-linked poly(vinyl pyrrolidone) (PVP) and poly(ethylene oxide)(PEO)/PVP electrospun nanofibrous mats containing complex-bound iodine have been studied. FT-IR spectroscopy analyses have proved that coordination of molecular iodine with carbonyl group and nitrogen atom of pyrrolidone rings of the PVP chains has taken place. The distribution of iodine along the fibers is uniform as revealed by X-ray mapping. The microbiological tests have demonstrated that the iodine complex-containing electrospun mats are highly effective against the Gram-positive bacterium Staphylococcus aureus, the Gram-negative bacterium Escherichia coli and the fungus Candida albicans. Comparison with iodine complex-containing films has shown that the iodine complex-containing nanofibers exhibit a higher killing rate than the films against bacteria E. coli. SEM observations showed that PVP-iodine nanofibrous mats inhibit the adhesion of bacteria S. aureus. These characteristic features make the electrospun iodine-containing nanofibers good candidates for wound-dressing materials.

[Ophthalmological symptoms in patients with giant pituitary adenomas].

In patients with giant pituitary adenomas, visual dysfunction is a common complaint and the chiasmatic syndrome is the major clinical sign of the disease. Reduced vision should be also considered to be a major factor that influences the quality of life. Early and timely diagnosis of a tumor in the absence of its extracellular growth makes it possible to improve visual function in the postoperative period and the quality of life.

[Invasive giant pituitary adenomas].

7 cases of pituitary adenomas with the pronounced signs of invasive growth, confirmed by MRI, clinical and intraoperative observations. Invasive growth of adenomas was related to neither the structure of the tumor, the degree of anaplasia, or the nature of a generated hormone.

Dynamic changes in nicotinamide pyridine dinucleotide content in normal human epidermal keratinocytes and their effect on retinoic acid biosynthesis.

The function of many enzymes that regulate metabolism and transcription depends critically on the nicotinamide pyridine dinucleotides. To understand the role of NAD(P)(H) in physiology and pathophysiology, it is imperative to estimate both their amount and ratios in a given cell type. In human epidermis and in cultured epidermal keratinocytes, we found that the total dinucleotide content is in the low millimolar range. The dinucleotide pattern changes during proliferation and maturation of keratinocytes in culture. Differences in the concentrations of NAD(P)(H) of 1.5- to 12-fold were observed. This resulted in alteration of the NAD(P)H/NAD(P) ratio, which could impact the differential regulation of both transcriptional and metabolic processes. In support of this notion, we provide evidence that the two-step oxidation of retinol to retinoic acid, a nuclear hormone critical for epidermal homeostasis, can be regulated by the relative physiological amounts of the pyridine dinucleotides.

Morphological variability and cell-wall deficiency in Mycobacterium tuberculosis 'heteroresistant' strains.

SETTING: Sofia State Hospital for Tuberculosis Treatment, Bulgaria. OBJECTIVE: To investigate the morphology of two 'heteroresistant' clinical isolates and one non-heteroresistant isolate, all isolated from newly diagnosed tuberculosis (TB) patients, as well as the reference strain H37Rv. Heteroresistant isolates contained clonally-related sensitive and drug-resistant organisms which could subsequently be separated using drug-containing primary cultures and had been isolated from patients originally diagnosed with susceptible TB by the 1% proportion method. Mycobacterial cultures were evaluated by transmission electron microscopy after 25 days of cultivation in Dubos broth. RESULTS: In contrast to H37Rv and the non-heteroresistant isolate, the bacterial populations in both heteroresistant isolates demonstrated distinct pleomorphic variability and coexistence of both classical and cell-wall deficient forms. Electron micrographs of mutants resistant to streptomycin and isoniazid showed predominance of atypical granular L-forms, which formed L-type colonies on Dubos agar. CONCLUSION: The L-form transformation processes, observed both in clinical heteroresistant isolates containing mixed populations of Mycobacterium tuberculosis organisms with different resistance gene genotypes and in the isolated resistant (mutant) clones, indicate a possible link between resistance and cell-wall deficient L-phase states and suggest one of the possible mechanisms by which resistant mutants are able to survive in vivo.

Intranasal application of lentinan enhances bactericidal activity of rat alveolar macrophages against Mycobacterium tuberculosis.

To study the in vivo interaction between lentinan-stimulated alveolar macrophages and Mycobacterium tuberculosis were used rats intranasally infected with 2 x 10(8) CFU. Before infection animals were treated intranasally with lentinan at a dose of 1 mg/kg (administrated three times at 2 days periods). Samples of broncho-alveolar lavage fluid were obtained from rats at different intervals -3, 24 and 72 h after infection. The process of phagocytosis in vivo was observed by light and electron microscopy, as well as acid phosphatase cytochemistry methods. Bactericidal activity of alveolar macrophages following the same intranasal installation of lentinan was assessed by in vitro "killing" ability test against M. tuberculosis. Nitrite production by lentinan activated alveolar macrophages was measured 24 h after ex vivo culture of these cells. It was demonstrated that lentinan induces high level of alveolar macrophage activation manifested through enhanced bactericidal effect against M. tuberculosis, which correlates with the induction of reactive nitrogen intermediates, increased activity of lysosomal enzymes (acid phosphatase), and with effective phagolysosomal fusion followed by destruction of Mycobacteria.

Properties of Yersinia enterocolitica and Yersinia pseudotuberculosis in red blood cell concentrate of different ABO groups during 30-day storage at 4 degrees C.

OBJECTIVE: The present study aimed to investigate the psychrophilic properties of Yersinia enterocolitica and Yersinia pseudotuberculosis as contaminants of donated blood. METHODS: Bags with red blood cell concentrates (RBCCs) of A, B, and AB blood groups were inoculated with a bacterial suspension of Y. enterocolitica (0 : 3 and 0 : 8) and Y. pseudotuberculosis (serovars I and III) and stored at 4 degrees C for 30 days. Bacterial growth was measured at selected intervals after inoculation. Initial strains and their subcultures (isolated after 30 days' incubation at 4 degrees C) were tested for glycolytic activity and susceptibility to the bactericidal action of human serum. RESULTS: It was found that all strains grew well in the RBCCs of A, B, and AB groups. After incubation at 4 degrees C they increased their glycolytic activity and became more sensitive to the killing ability of human serum. CONCLUSIONS: The prolonged storage of contaminated Y. enterocolitica and Y. pseudotuberculosis RBCCs at 4 degrees C induces bacterial multiplication to high levels and stimulates glycolytic activity of bacterial cells.

[Ultrasonic examinations of thyroid newgrowths]. / Ul'trazvukovye metodiki issledovaniia ob''emnykh obrazovanii shchitovidnoi zhelezy.

The results of complex ultrasonic, histological and cytological examination of 148 patients with newgrowths thyroid were analyzed. Grey-scale mode, colored and energy Doppler scanning, three-dimensional reconstruction of vessels were used. A new classification of nodular vascular pattern types based on 3-D reconstruction is proposed. The majority of malignant formations were characterized by perinodular hypervascularization and internodular hypovascularization (82.4%), adenomas--by peri- and intranodular hypervascularization (88.9%), colloid nodes--by perinodular hypervascularization (85.5%), the blood flow was absent in simple cysts (85.3%). It was established that complex of these methods increases informative value of ultrasonic examination in diagnosis of thyroid nodular diseases: in malignant neoplasms the sensitivity is 92.9%, specificity--86.1%, diagnostic accuracy--86.9%, in adenomas--93.4%, 79.2% and 82%, in colloid nodes--75.7%, 84.1% and 79.8%, respectively. In cystic masses diagnostic value of Doppler methods didn't change: sensitivity was 90.4%, specificity--63%, diagnostic accuracy--80.5%. The combined ultrasonic examination provides a differential approach to treatment of patients with thyroid nodular masses and decreases the number of unjustified surgical interventions.

Comparison of three methods to find the vapor activity of a hydration step.

The formation of a stochiometric salt hydrate takes place at a well defined vapor activity. We have compared three methods to measure this vapor activity. In two of the methods we used a sorption balance in step mode and in ramp mode, respectively, and in one method we used a newly developed sorption microcalorimeter. The tests were made with the formation of morphine sulphate pentahydrate from its dihydrate at 25 degrees C. With all three methods this transition was found to take place at a vapor activity close to 0.21.

Interaction of alveolar macrophages with Staphylococcus aureus and induction of microbial L-forms during infection in rats.

In vivo cell interactions between Staphylococcus aureus and rat alveolar macrophages were investigated after intranasal inoculation during a 30-days period of examination. Some dynamic characteristics of microorganisms in the macrophages were examined by electron microscopy and acid phosphatase cytochemistry. It was found that at earlier infection intervals (days 3 and 7) the ingested cocci were sequestered in phagosomes and phagolysosomes and later many of the microbial cells were digested. An interesting finding was the intracellular appearance of cell wall-defective forms (L-forms) of S. aureus at later intervals (days 14 and 30 after challenge). Infection kinetics were evaluated by isolation and enumeration of colony-forming units of S. aureus from bronchoalveolar fluid and by assessment of blood and bronchoalveolar total and differential leukocyte counts. The results indicate that induction and survival of S. aureus L-forms may occur spontaneously in vivo. This phenomenon could explain some of the mechanisms, provoking the latent and relapsing lung infections.

Complex interactions between epidermal POU domain and activator protein 1 transcription factors regulate the expression of the profilaggrin gene in normal human epidermal keratinocytes.

The human profilaggrin gene is expressed in the granular layer during the late stages of the epidermal differentiation. The proximal promoter region of the gene confers high levels of keratinocyte-specific transcription via interactions with c-Jun/c-Fos heterodimers. Here we provide evidence for another level of complexity in the regulation of the profilaggrin promoter activity. The POU domain proteins Oct1, Skn1a/i, and Oct6, which are abundantly expressed in the epidermal cells, act to both stimulate and repress transcription in a general and a cell type-specific mode. While binding to specific recognition elements within the promoter region, they exert their effects by either stimulating or antagonizing the c-Jun-dependent activity of the promoter. The response of the promoter to forced expression of the POU domain proteins reflects the effect of these transcription factors on the endogenous profilaggrin mRNA synthesis and suggests that the latter requires a fine balance in the amounts and the activities of the individual activator protein 1 and POU domain proteins.

Atypical behaviour and survival of Streptococcus pyogenes L forms during intraperitoneal infection in rats.

Groups of rats were injected intraperitoneally with cell wall-deficient (L) forms of Streptococcus pyogenes, with their parental (S) forms, as well as with a combined inoculum of both forms (S+L). Peritoneal exudate samples were harvested on days 1, 3, 7, 15 and 30 after challenge and were investigated by microbiological, electron microscopic, cytometric and biochemical methods. Parental S forms were isolated from peritoneal exudate samples up to day 15 post infection, while L form cultures were isolated until the end of the examined interval. Electron microscopic examination revealed continuous adhesion of L forms on the macrophage surface as well as intracellular persistence inside them. It was demonstrated that the intraperitoneal inflammatory response to L form infection was higher than to the other infections and the monocyte-macrophage populations were predominant. The established atypical behaviour and long survival of S. pyogenes L forms in the rat's peritoneum could explain some of the mechanisms of the pathogens' persistence as well as the reasons for chronic streptococcal infections.

Cell-wall-deficient forms of Staphylococcus aureus as lung pathogens: an ultrastructural study.

The interaction between stable protoplast L forms of Staphylococcus aureus and the alveolar wall of infected rats was observed in the course of experimental pulmonary infection (days 3, 7, 14, and 30 p.i.). The L forms were successfully cultivated from bronchoalveolar lavage samples taken throughout the tested interval. The ultrastructural results demonstrated the ability of the L forms to invade the alveolar wall where they established, grew and reproduced mainly in the interstitium. The infection caused lung lesions: granulomas, focal fibroses and destruction of type I alveolar epithelial cells.

Persistence and in vivo effects of Yersinia enterocolitica 0:3 endotoxin in rats.

In vivo effects of Yersinia enterocolitica 0:3 lipopolysaccharide (prepared from bacteria grown at 25 degrees C and 37 degrees C) were investigated after intraperitoneal (i.p.) and intraarticular (i.a.) injection in rats during 30 days of examination. The persistence of endotoxin in the peritoneal and the synovial cavities was demonstrated by the immunofluorescence technique. Peritoneal and synovial exudative cell infiltration, as well as changes in some parameters (glycolytic and acid phosphatase activity, and killing ability of peritoneal cells; lactate dehydrogenase concentration in synovial fluid) were studied. The results indicated that endotoxin could persist longer in the synovial than in the peritoneal cavity.