Extended Prophylactic Effect of N-tert-Butyl-α-phenylnitron against Oxidative/Nitrosative Damage Caused by the DNA-Hypomethylating Drug 5-Azacytidine in the Rat Placenta

Antioxidant N–tert–Butyl–α–phenylnitron (PBN) partly protected embryos from the negative effects of a DNA demethylating drug 5-azacytidine during pregnancy. Our aim was to investigate PBN’s impact on the placenta. Fischer rat dams were treated on gestation days (GD) 12 and 13 by PBN (40 mg/kg), followed by 5azaC (5 mg/kg) after one hour. Global methylation was assessed by pyrosequencing. Numerical density was calculated from immunohistochemical expression in single cells for proliferating (PCNA), oxidative (oxoguanosine) and nitrosative (nitrotyrosine) activity. Results were compared with the PBN-treated and control rats. PBN-pretreatment significantly increased placental weight at GD15 and GD20, diminished by 5azaC, and diminished apoptosis in GD 20 placentas caused by 5azaC. Oxoguanosine expression in placentas of 5azaC-treated dams was especially high in the placental labyrinth on GD 15, while PBN-pretreatment lowered its expression on GD 15 and GD 20 in both the labyrinth and basal layer. 5azaC enhanced nitrotyrosine level in the labyrinth of both gestational stages, while PBN-pretreatment lowered it. We conclude that PBN exerted its prophylactic activity against DNA hypomethylating agent 5azaC in the placenta through free radical scavenging, especially in the labyrinthine part of the placenta until the last day of pregnancy.

Pityriasis lichenoid-like mycosis fungoides

Sir, Mycosis fungoides is a primary cutaneous T–cell lymphoma, secondary clonal proliferation of mature skin-homing T cells, mostly CD4-positive, with a predilection for involving the epidermis. It is an indolent lymphoma that progresses over several years and represents 50% of primary cutaneous T-cell lymphomas [1]. Its clinical presentation is variable, thus leading to several clinical variants. Herein, we describe a rare variant of mycosis fungoides: pityriasis lichenoid-like mycosis fungoides. A 45-year-old female was referred to our department with a papular rash evolving for the last year without regression. The patient had a history of breast carcinoma in complete remission for two years. A clinical examination revealed erythematous, scaly, non-itchy papules covering the entire body but sparing the face (Figs. 1 and 2). There was no scalp involvement or associated lymphadenopathy. Based on the clinical presentation, the suggested diagnosis was pityriasis lichenoid. A histological examination revealed Pautrier’s microabscesses, atypical lymphocyte infiltration along the basal layer and papillary dermis, and prominent epidermotropism (Fig. 3). There was pilotropism without mucin. Besides, hyperkeratosis with focal parakeratosis and perivascular infiltrate were noted. An immunohistochemical analysis revealed infiltrates of T cells expressing CD3, CD2, CD5, and a predominance of CD4-positive T cells in the epidermis compared to CD8-positive T cells. CD7 and CD30 were, however, negative. These findings were consistent with pityriasis lichenoid-like mycosis fungoides. The patient was classified as a IB stage and received UVB phototherapy with good progress.

Comparative characteristics of different methods of treatment of microsporia

Microsporia is the most common disease of fungal etiology, most often caused by the pathogen Microsporum canis. Treatment of this disease requires a comprehensive approach, because the disease is dangerous to humans. Therefore, in the treatment of microspores, it is important not only to carry out therapeutic measures, but also to prevent the spread of the pathogen of the fungus in the environment and increase the immune status of the organism in the fight against infection. The main source of infection is cats. In order to determine the effectiveness of treatment of microsporia by various methods, studied of the blood and skin of guinea pigs infected with the pathogen M. canis. Sick animals were divided into three groups. The first group was treated with the systemic antifungal itraconazole and topical treatment with a solution of clotrimazole. Treatment of the second group was performed with a topical antifungal agent (1% solution of clotrimazole) with vaccination with the antifungal vaccine “Vakderm”. For the third group, the developed drugs were used – antifungal agent “Micromar” and immunostimulant “Biogluk”. During treatment, hematological and immunological blood researches and histological skin examinations were performed. During treatment with antifungal drugs (itraconazole and clotrimazole) the number of leukocytes decreases from 11.13 ± 0.72 to 7.13 ± 0.22, rod-shaped neutrophils from 15.76 ± 1.29 to 5.50 ± 0.76, and segmental increases from 12.17 ± 1.47 to 24.17 ± 2.27, decreases ESR from 5.67 ± 0.67 to 2.33 ± 0.42, which occurs when inhibiting the inflammatory response of the organism to infection. Thrombocytopenia (from 231.17 ± 7.60 to 184.33 ± 7.65) and eosinophilia (from 2.70 ± 0.73 to 7.33 ± 1.33) are also noted. There is a slight increase in T-helpers and a decrease in T-suppressors. Histologically, the infiltration of the dermis by histiocytes and eosinophils under skin persists. In the treatment of microsporia by treatment with 1 % solution of clotrimazole and vaccination with the vaccine “Vakderm” the results of studies showed that the number of leukocytes decreases from 11.13 ± 0.72 to 5.35 ± 0.31 (P < 0.01), rod-shaped neutrophils from 15.76 ± 1.29 to 7.67 ± 0.56, and segmental increases from 12.17 ± 1.47 to 22.17 ± 0.91 (P < 0.001), decreases). The number of T-helpers is increasing. The histological picture on day 7 is characterized by hyperkeratosis, and on day 14 the hyperemia of the basal layer of the epidermis persists. When using the antifungal drug “Micromar” and immunostimulant “Biogluk” the results of studies showed that the number of leukocytes decreases from 11.13 ± 0.72 to 6.95 ± 0.10, rod-shaped neutrophils from 15.76 ± 1.29 to 6.17 ± 0.65, and segmental increases from 12.17 ± 1.47 to 22.00 ± 0.86, decreases ESR from 5.67 ± 0.67 to 2.17 ± 0.31. Increases the number of T-helpers and the number of natural killers and T-suppressors is gradually decreasing. Histological changes are presented in the form of dilation of blood vessels and visualization of single erythrocytes in the dermis on day 14 of treatment.

Nfatc1’s Role in Mammary Epithelial Morphogenesis and Basal Stem/progenitor Cell Self-renewal

Mammary gland is an outstanding system to study the regulatory mechanisms governing adult epithelial stem cell activity. Stem cells in the basal layer of the mammary gland fuel the morphogenesis and regeneration of a complex epithelial network during development and upon transplantation. The self-renewal of basal stem/progenitor cells is subjected to regulation by both cell-intrinsic and extrinsic mechanisms. Nfatc1 is a transcription factor that regulates breast tumorigenesis and metastasis, but its role in mammary epithelial development and stem cell function has not been investigated. Here we show that Nfatc1 is expressed in a small subset of mammary basal epithelial cells and its epithelial-specific deletion results in mild defects in side branching and basal-luminal cell balance. Moreover, Nfatc1-deficient basal cells exhibit reduced colony forming ability in vitro and somewhat compromised regenerative potential upon transplantation. Thus, our study provides evidence for a detectable yet non-essential role of Nfatc1 in mammary epithelial morphogenesis and basal stem/progenitor cell self-renewal.

Ultrastructural Analysis of Rehydrated Human Donor Corneas After Air-Drying and Dissection by Femtosecond Laser

Purpose: To evaluate the efficiency of femtosecond laser (FSL) incision of rehydrated human donor corneas after air-drying and its effects on corneal structure.Methods: We compared the rehydrated and fresh-preserved corneas by microscopy following Victus-Tecnolas FSL treatment for straight-edge anterior lamellar keratoplasty (ALK). The corneas were dehydrated at room temperature under a laminar-flow hood.Results: To obtain the horizontal cut in rehydrated corneas, we increased the FSL pulse energy to 1.2 μJ from 0.80 μJ applied for the fresh corneas and obtained a clear-cut separation of the lamellar lenticule cap from the corneal bed. Light microscopy showed regular arrangement of stromal collagen lamellae, with spaces in between the fibers in the corneal stroma in the fresh and the rehydrated corneas, but the uppermost epithelial layers in the rehydrated corneas were lost. Transmission electron microscopy (TEM) revealed no signs of thermal or mechanical damage to the corneal structure. The epithelial basal membrane and Bowman's layer maintained their integrity. The epithelial basal layer and cells were separated by large spaces due to junction alteration in the rehydrated corneas. There were gaps between the lamellar layers in the stroma, especially in the rehydrated corneas. Keratocytes displayed normal structure in the fresh corneas but were devoid of microorganules in the rehydrated corneas. Minor irregularities were observed in the vertical incision and the horizontal stroma appeared smooth on scanning electron microscopy.Conclusion: The corneal stroma of rehydrated corneas maintained morphology and integrity, while corneal cellular components were generally altered. When corneas are intended for FSL-assisted ALK, effective stromal bed incision is best achieved at a laser power higher than that currently adopted for fresh corneas.

BMSCs and Pectin-Based E2-Loaded Microcapsules with Injectable Pectin-Pluronic® F-127 Scaffolds for Mouse Endometrial Regeneration Application

Background Uterine endometrium is a highly dynamic tissue which consists of a basal layer and a functional layer. Bone marrow-derived mesenchymal stem cells (BMSCs) have been recognized as new candidates for the treatment of serious endometrial injuries. However, due to the local microenvironment of damaged endometrium, transplantation of BMSCs yielded disappointing results with respect to survival, attachment, differentiation, and proliferation. Methods Pectin-Pluronic® F-127 scaffolds were fabricated. E2 was encapsulated into the W/O/W microspheres to construct pectin-based E2-loaded microcapsules (E2 MPs). The BMSCs/E2 MPs/scaffolds system was then injected into the uterine cavity of mouse endometrial injury model. Furthermore, the mechanism of E2 in promoting the repair of endometrial injury was also investigated. Result Pectin-Pluronic® F-127 scaffolds could provide three-dimensional architecture for the attachment, growth, and migration of BMSCs. E2 MPs has the potential to serve as a long-term reliable source of E2 for endometrial regeneration. At four weeks after transplantation, it was demonstrated that the system increased proliferative abilities of uterine endometrial cells, facilitated microvasculature regeneration, and restored the ability of endometrium to receive an embryo, suggesting that the BMSCs/E2 MPs/scaffolds system is a promising treatment option for endometrial regeneration. Exosomes are critical paracrine mediators that act as biochemical cues to direct stem cell differentiation. In this study, it was found that the expression of endometrial epithelial cells (EECs) markers was up-regulated in BMSCs treated by exosomes secreted from endometrial stromal cells (ESCs-Exos). Exosomes derived from E2-stimulated ESCs further promoted the expression level of EECs markers in BMSCs, suggesting exosomes released from ESCs by E2 stimulation could enhance the differentiation efficiency of BMSCs. Conclusion The BMSCs/E2 MPs/scaffolds therapeutic strategy may be beneficial in the treatment of severely damaged endometrium. Exosomes derived from ESCs play paracrine roles in endometrial regeneration stimulated by E2, potentially modulating the differentiation of BMSCs.

Coupled dynamics and evolution of primordial and recycled heterogeneity in Earth’s lower mantle

The nature of compositional heterogeneity in Earth’s lower mantle remains a long-standing puzzle that can inform about the long-term thermochemical evolution and dynamics of our planet. Here, we use global-scale 2D models of thermo- chemical mantle convection to investigate the coupled evolution and mixing of (intrinsically-dense) recycled and (intrinsically- strong) primordial heterogeneity in the mantle. We explore the effects of ancient compositional layering of the mantle, as motivated by magma-ocean solidification studies, and of the physical parameters of primordial material. Depending on these physical parameters, our models predict various regimes of mantle evolution and heterogeneity preservation over 4.5 Gyrs. Over a wide parameter range, primordial and recycled heterogeneity are predicted to co-exist with each other in the lower mantle of Earth-like planets. Primordial material usually survives as mid-to-large scale blobs (or streaks) in the mid-mantle, around 1000-2000 km depth, and this preservation is largely independent on the initial primordial-material volume. In turn, recycled oceanic crust (ROC) persists as large piles at the base of the mantle and as small streaks everywhere else. In models with an additional dense FeO-rich layer initially present at the base of the mantle, the ancient dense material partially survives at the top of ROC piles, causing the piles to be compositionally stratified. Moreover, the addition of such an ancient FeO-rich basal layer significantly aids the preservation of the viscous domains in the mid-mantle. Finally, we find that primordial blobs are commonly directly underlain by thick ROC piles, and aid their longevity and stability. Based on our results, we propose an integrated style of mantle heterogeneity for the Earth, involving the preservation of primordial domains along with recycled piles. This style has important implications for early Earth evolution, and has the potential of reconciling geophysical and geochemical discrepancies on present-day lower-mantle heterogeneity.

Comfortable and dermatological effects of hot spring bathing provide demonstrative insight into improvement in the rough skin of Capybaras

The purpose of this study was to clarify dermatologically the favorable effects of hot spring bathing on the rough skin in Capybaras. Non-volcanic hot springs used in this study showed alkaline quality of water (pH 9.3), containing sodium and chloride ions. The normal skin in Capybaras was characterized by the presence of relatively thick epidermis with mild alkaline state (pH 8.26). The dorsal skin had melanin granules in the basal layer. Their rough skin affected in the Japanese cold winter was improved by daily bathing in an alkaline hot spring. The skin properties returned to the normal skin conditions (moisture, melanin and erythema values) observed in the summer. The facial expression mainly changes in the eyes was scored to evaluate comfortable status. The comfortable status during hot spring bathing significantly increased as compared with that observed before bathing (p < 0.01). The thermography revealed a heat retention effect of body temperature after hot spring bathing for 30 min. In conclusion, this study demonstrates that hot spring had significantly comfortable and dermatological effects on the basis of evaluation for the skin and body conditions in Capybaras.

Clinico-echographic manifestation of endometritis in cows

Purpose: to study the clinical and echographic manifestation of endometritis.Materials and methods. Clinical and echographic research was carried out on 19 red-and-white cows, in the conditions of the agricultural farm of the Druzhba breeding plant, Pavlovsky district, Voronezh region on the 30-32th day after calving using the Easi-Scan scanner from BCF Technology Ltd, Scotland. Before ultrasound examination to establish the diagnosis, all animals were examined, transrectal palpation of the uterus. Simultaneously with the ultrasound examination, a cytological study of the uterine mucosa was carried out. For this purpose, a probe was constructed from a metal catheter intended for artificial insemination of cows with sequins by the deep cervical method with rectal fixation of the cervix; a cytobrush of the Juno probe was attached to the tip of the instrument. To confirm the diagnosis, a laboratory study of cervical mucus was additionally carried out by the express method according to Whiteside, modified by N. I. Polyantsev and Yu. N. Popov.Results. According to the results of rectal examination and ultrasound diagnostics, the cows were divided into three groups: clinically healthy, animals with clinically pronounced chronic endometritis and with latent endometritis. In the group of healthy animals, individual superficial and vacuolated intermediate cells (6.17 ± 0.51) and single neutrophils (2.31 ± 0.32) were detected in smears. In the group of clinically healthy cows, no gram-positive coccobacillary microorganisms and cells of the basal layer of the uterine mucosa were detected. In the second group, in animals with clinically pronounced chronic endometritis, a large number of segmented neutrophils, lymphocytes and a large number of cocci were detected in smears. Thus, in one visual field, the number of gram-positive cocobacillary microorganisms fluctuated on average 764.45 ± 4.56 microbial bodies, while the number of neutrophils increased in comparison with clinically healthy animals by 20.84 times and averaged 48.14 ± 2 in the group. , 91, the percentage of the score averaged around 12.1%. Simultaneously with an increase in the number of leukocytes in animals, the number of epithelial cells of the uterine mucosa increased by 2.21 times, while individual basal cells were detected simultaneously with superficial and intermediate cells (0.75 ± 0.48). In smears obtained from animals of the third group of cows, there was a significant decrease in the number of coccobacillary microorganisms to 75-360 in one visual field in comparison with similar results in the group with a clinically pronounced form of chronic endometritis. The indicator for segmented neutrophils was 8.05 times higher than in clinically healthy cows, while in relation to clinically sick cows by 61.4% and amounted to 18.60 ± 2.23. In terms of the number of superficial, vacuolated intermediate and basal cells in sick animals with chronic clinically pronounced endometritis and latent endometritis, no significant changes were revealed (1.08 times), while in relation to clinically healthy animals, these indicators were 2.38 times higher. The degree of variation in the number of epithelial cells in the group of cows with latent chronic endometritis was insignificant, which indicates the stability of the trait.Conclusion. Ultrasound scanning allows you to identify the latent form of endometritis and establish degenerative changes in the tissues of the uterus. Ultrasound examination in cows in establishing a diagnosis - endometritis should be the decisive diagnostic method.