Repetitive hyperbaric oxygenation attenuates reactive astrogliosis and suppresses expression of inflammatory mediators in the rat model of brain injury.

The exact mechanisms by which treatment with hyperbaric oxygen (HBOT) exerts its beneficial effects on recovery after brain injury are still unrevealed. Therefore, in this study we investigated the influence of repetitive HBOT on the reactive astrogliosis and expression of mediators of inflammation after cortical stab injury (CSI). CSI was performed on male Wistar rats, divided into control, sham, and lesioned groups with appropriate HBO. The HBOT protocol was as follows: 10 minutes of slow compression, 2.5 atmospheres absolute (ATA) for 60 minutes, and 10 minutes of slow decompression, once a day for 10 consecutive days. Data obtained using real-time polymerase chain reaction, Western blot, and immunohistochemical and immunofluorescence analyses revealed that repetitive HBOT applied after the CSI attenuates reactive astrogliosis and glial scarring, and reduces expression of GFAP (glial fibrillary acidic protein), vimentin, and ICAM-1 (intercellular adhesion molecule-1) both at gene and tissue levels. In addition, HBOT prevents expression of CD40 and its ligand CD40L on microglia, neutrophils, cortical neurons, and reactive astrocytes. Accordingly, repetitive HBOT, by prevention of glial scarring and limiting of expression of inflammatory mediators, supports formation of more permissive environment for repair and regeneration.

Discriminatory ability of fractal and grey level co-occurrence matrix methods in structural analysis of hippocampus layers.

Fractal and grey level co-occurrence matrix (GLCM) analysis represent two mathematical computer-assisted algorithms that are today thought to be able to accurately detect and quantify changes in tissue architecture during various physiological and pathological processes. However, despite their numerous applications in histology and pathology, their sensitivity, specificity and validity regarding evaluation of brain tissue remain unclear. In this article we present the results indicating that certain parameters of fractal and GLCM analysis have high discriminatory ability in distinguishing two morphologically similar regions of rat hippocampus: stratum lacunosum-moleculare and stratum radiatum. Fractal and GLCM algorithms were performed on a total of 240 thionine-stained hippocampus micrographs of 12 male Wistar albino rats. 120 digital micrographs represented stratum lacunosum-moleculare, and another 120 stratum radiatum. For each image, 7 parameters were calculated: fractal dimension, lacunarity, GLCM angular second moment, GLCM contrast, inverse difference moment, GLCM correlation, and GLCM variance. GLCM variance (VAR) resulted in the largest area under the Receiver operating characteristic (ROC) curve of 0.96, demonstrating an outstanding discriminatory power in analysis of stratum lacunosum-moleculare (average VAR equaled 478.1 ± 179.8) and stratum radiatum (average VAR of 145.9 ± 59.2, p < 0.0001). For the criterion VAR ≤ 227.5, sensitivity and specificity were 90% and 86.7%, respectively. GLCM correlation as a parameter also produced large area under the ROC curve of 0.95. Our results are in accordance with the findings of our previous study regarding brain white mass fractal and textural analysis. GLCM algorithm as an image analysis method has potentially high applicability in structural analysis of brain tissue cytoarcitecture.

Melatonin: a "Higgs boson" in human reproduction.

As the Higgs boson could be a key to unlocking mysteries regarding our Universe, melatonin, a somewhat mysterious substance secreted by the pineal gland primarily at night, might be a crucial factor in regulating numerous processes in human reproduction. Melatonin is a powerful antioxidant which has an essential role in controlling several physiological reactions, as well as biological rhythms throughout human reproductive life. Melatonin, which is referred to as a hormone, but also as an autocoid, a chronobiotic, a hypnotic, an immunomodulator and a biological modifier, plays a crucial part in establishing homeostatic, neurohumoral balance and circadian rhythm in the body through synergic actions with other hormones and neuropeptides. This paper aims to analyze the effects of melatonin on the reproductive function, as well as to shed light on immunological and oncostatic properties of one of the most powerful hormones.

Application of fractal and grey level co-occurrence matrix analysis in evaluation of brain corpus callosum and cingulum architecture.

This aim of this study was to assess the discriminatory value of fractal and grey level co-occurrence matrix (GLCM) analysis methods in standard microscopy analysis of two histologically similar brain white mass regions that have different nerve fiber orientation. A total of 160 digital micrographs of thionine-stained rat brain white mass were acquired using a Pro-MicroScan DEM-200 instrument. Eighty micrographs from the anterior corpus callosum and eighty from the anterior cingulum areas of the brain were analyzed. The micrographs were evaluated using the National Institutes of Health ImageJ software and its plugins. For each micrograph, seven parameters were calculated: angular second moment, inverse difference moment, GLCM contrast, GLCM correlation, GLCM variance, fractal dimension, and lacunarity. Using the Receiver operating characteristic analysis, the highest discriminatory value was determined for inverse difference moment (IDM) (area under the receiver operating characteristic (ROC) curve equaled 0.925, and for the criterion IDM≤0.610 the sensitivity and specificity were 82.5 and 87.5%, respectively). Most of the other parameters also showed good sensitivity and specificity. The results indicate that GLCM and fractal analysis methods, when applied together in brain histology analysis, are highly capable of discriminating white mass structures that have different axonal orientation.

Hyperbaric oxygenation improves locomotor ability by enhancing neuroplastic responses after cortical ablation in rats.

OBJECTIVE: To investigate whether hyperbaric oxygenation (HBO) can improve the recovery of motor functions in rats after suction ablation of the right sensorimotor cortex. METHODS: The experimental paradigm implies the following groups: Control animals (C), Control + HBO (CHBO), Sham controls (S), Sham control + HBO (SHBO), Lesion group (L), right sensorimotor cortex was removed by suction, Lesion + HBO (LHBO). Hyperbaric protocol: pressure applied 2.5 atmospheres absolute, for 60 minutes, once a day for 10 days. A beam walking test and grip strength meter were used to evaluate the recovery of motor functions. Expression profiles of growth-associated protein 43 (GAP43) and synaptophysin (SYP) were detected using immunohistochemistry. RESULTS: The LHBO group achieved statistically superior scores in the beam walking test compared to the L group. Additionally, the recovery of muscle strength of the affected hindpaw was significantly enhanced after HBO treatment. Hyperbaric oxygenation induced over-expression of GAP43 and SYP in the neurons surrounding the lesion site. CONCLUSIONS: Data presented suggest that hyperbaric oxygen therapy can intensify neuroplastic responses by promoting axonal sprouting and synapse remodelling, which contributes to the recovery of locomotor performances in rats. This provides the perspective for implementation of HBO in clinical strategies for treating traumatic brain injuries.

Hyperbaric oxygenation alters temporal expression pattern of superoxide dismutase 2 after cortical stab injury in rats.

AIM: To evaluate the effect of hyperbaric oxygen therapy (HBOT) on superoxide dismutase 2 (SOD2) expression pattern after the cortical stab injury (CSI). METHODS: CSI was performed on 88 male Wistar rats, divided into control, sham, lesioned, and HBO groups. HBOT protocol was the following: pressure applied was 2.5 absolute atmospheres, for 60 minutes, once a day for consecutive 3 or 10 days. The pattern of SOD2 expression and cellular localization was analyzed using real-time polymerase chain reaction, Western blot, and double-label fluorescence immunohistochemistry. Neurons undergoing degeneration were visualized with Fluoro-Jade®B. RESULTS: CSI induced significant transient increase in SOD2 protein levels at day 3 post injury, which was followed by a reduction toward control levels at post-injury day 10. At the same time points, mRNA levels for SOD2 in the injured cortex were down-regulated. Exposure to HBO for 3 days considerably down-regulated SOD2 protein levels in the injured cortex, while after 10 days of HBOT an up-regulation of SOD2 was observed. HBOT significantly increased mRNA levels for SOD2 at both time points compared to the corresponding L group, but they were still lower than in controls. Double immunofluorescence staining revealed that 3 days after CSI, up-regulation of SOD2 was mostly due to an increased expression in reactive astrocytes surrounding the lesion site. HBOT attenuated SOD2 expression both in neuronal and astroglial cells. Fluoro-Jade®B labeling showed that HBOT significantly decreased the number of degenerating neurons in the injured cortex. CONCLUSION: HBOT alters SOD2 protein and mRNA levels after brain injury in a time-dependent manner.

An immunological insight into premature ovarian failure (POF).

Premature ovarian failure (POF), a serious life-changing condition that affects young women, remains an enigma and the researchers' challenge. The term POF generally describes a syndrome of gonadal failure before the age of 40, characterized by amenorrhea, sex steroid deficiency and elevated levels of gonadotropins. Infertility and psychological stress are common consequences of this entity the prevalence of which is 0.9-3%. The known cause of this condition includes: genetic aberrations, autoimmune ovarian damage, iatrogenic and environmental factors, although in majority of cases the underlying cause is not identified. For many women in whom the cause of ovarian failure is unknown, autoimmunity may be the pathogenic mechanism. There is currently evidence that some cases of POF are due to faulty recognition of self in the ovary by the immune system, possibly provoked by genetic or environmental factors initiating such immune response. Numerous evidence, including association with multiple autoimmune endocrine disorders, clinical reversibility, transitory estrogen deficiency, histological and immunological features and the demonstration of circulating ovarian antibodies in serum samples from women with POF, have suggested its immunological origin. We discuss the possible role of such an autoimmune process as a cause or consequence of POF including treatment strategies in POF patients.

[Hyperbaric oxygen therapy of angiopathic changes in patients with inherited gene imbalance].

INTRODUCTION: Phenotype match inherited by genes is in most cases present in monozygotic twins. Their phenotypic resemblance is unfortunately characterized by strong susceptibility for the development of chronic non-infectious diseases. One of the most common non-infectious chronic diseases that are phenotipically represented in twins is diabetes mellitus. Genetic imbalance is, in most cases, placed in 2, 3, 7, 8, 11, 12, 19 and 20 chromosomal pair of the human genome. CASE OUTLINE: This study describes a pair of monozygotic twins, aged 54, who were diagnosed for diabetes type 2 ten years earlier. The first patient had trophic changes of muscles and skin tissues of the lower limb, and a necrotic wound on his right leg tibial region with the claudication distance of 50 m. After arteriography, he was referred by a vascular surgeon for hyperbaric oxygen therapy (HBO). HBO protocol implied 70 min. application of 100% oxygen at 2.5 absolute atmospheres. After the first series of HBO therapies consisting of 20 HBO treatments, claudication was eliminated and the necrotic wound healed. Next, surgical aortofemoral bypass was done. During the second HBO treatment, his monozygotic twin brother presented with angiopathic changes due to diabetes. In both patients, biochemical parameters corresponded to the expected level for diabetes type 2 imbalance, and the localization of the chromosomal defect (placed on 3, 11 and 19 chromosomal pair) was also in accordance with the respective disorder. After they were included into next 10 HBO treatments, Doppler imaging of the major arteries of limbs revealed normal findings. CONCLUSION: Identical genetic impairment in monozygotic twins can lead to identical somatic changes with resultant consequences. HBO treatment of such patients associated with other therapeutic procedures (conducted by diabetologist, vascular surgeon and physiatrist) can postpone or prevent irreversible changes occurring due to blood vessel disorders.

Hyperbaric oxygenation as a possible therapy of choice for infertility treatment.

Endometrial sonographic and color doppler features can be used to predict the occurrence of pregnancy in natural or stimulated cycles. Implantation will usually only take place if the endometrium has reach a certain stage of vascularisation and development. The aim of this study was to evaluate endometrial development -- endometrial thickness and reflectivity , subendometrial, endometrial and uterine perfusion, after hyperbaric oxygenation, using transvaginal color doppler. During a three years period 32 women with unexplained infertility were entered into a randomised study. The patients were treated in multiplaced HAUX chamber at pressure of 2.3 ATA during 70 minutes, 7 days consecutively beginning with day 5th of menstrual cycle. The evaluation of effects of hyperbaric oxygen therapy was carried out by transvaginal color doppler sonography which was continuously used starting from 8th day of menstrual cycle until the ovulation in the cycles when the therapy was applied , one month before and one month after the therapy. Folliculometry in the cycles when hyperbaric oxygen therapy at 2.3 ATA was applied, indicated an excellent response of endometrium. Thickness of endometrium at the time of ovulation was 11.0 +/- 2.6 mm. Desirable quality of endometrium was significantly better in the cycle when HBO therapy had been applied (p< 0.001). The doppler flowmetry of the uterine arteries indicated that the uterine blood vessel resistance was slightly higher than expected. Mapping of subendometrial blood vessels in the cycles covered by hyperbaric oxygen therapy showed the intensive capillary network of endometrium with low resistance Ri< 0.45. The oxygen used under higher pressure -- oxygen as a drug , may have an extraordinary significance for better outcome of pregnancy implantation by improving endometrial receptivity. If endometrial receptivity is conditioned by adequate vascularisation and oxygenation, then hyperbaric oxygen therapy is the treatment of choice.

Relation between body mass and postmenopausal symptoms in surgically castrated women.

INTRODUCTION: Post-menopause androgens are the essential precursors for estrogen synthesis through their conversion in peripheral tissues. The main postmenopausal estrogen is estrone, which is also peripherally converted into estradiol. Concentrations of these two estrogens are directly proportional in circulation. We wanted to establish whether there is a direct correlation between body mass and postmenopausal symptoms in surgically castrated women. MATERIALS AND METHODS: We analyzed 30 patients with artificially induced menopause, 3-12 months after the operation (hysterectomy and bilateral adnexectomy). Prior to being put on hormone substitution therapy the following was obtained from the patients: Body Mass Index--BMI (kg/m(2)), measurement of estradiol (E(2)) by RIA method, and 12 subjective symptoms analyzed according to intensity and frequency. The relationship between BMI, E(2) level and subjective symptoms were assessed. RESULTS: By analyzing the correlation coefficient it was shown that there was statistically significant causal-consequential connection between the E(2) level and BMI (R(2)=0.1647, p<0.05). There is no strong correlation between BMI and symptoms (R(2)=0.004, p<0.05). However, E(2) level correlated with subjective symptoms (R(2)=0.2123, p<0.05). CONCLUSIONS: Estrogen production in postmenopause is dependent on the substrate availability, and thus with adiposity. Therefore, we were surprised by the lack of correlation between BMI and symptoms. This suggests that estrogen metabolism and biological effects are not primarily affected by BMI, and that expression of postmenopausal symptoms does not depend only on estrogen level.