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The gracilis myocutaneous flap is a versatile reconstructive option. Despite wide applicability, concerns are often raised regarding the unreliable vascular perfusion of the distal third of the flap’s skin paddle. This study presents our experience utilizing the pedicled gracilis myocutaneous flap for various oncologic defects to highlight the versatility, safety, and utility of this flap. A retrospective analysis was performed for patients who underwent reconstruction utilizing a gracilis myocutaneous flap over the course of 20 years (January 1998–June 2018). Postoperative outcomes were reviewed to determine the incidence of infection, dehiscence, seroma, or hematoma for both donor and recipient sites. Incidence of flap skin or muscle loss was also recorded. A total of 37 patients met our inclusion criteria, and data from 41 vertically oriented gracilis myocutaneous flaps was analyzed. A majority of our patients had a previously irradiated wound bed (73%). The overall donor site complication rate was 19.5%. The recipient site complication rate was 41.5% and included partial flap loss (14.6%). Partial flap loss included distal flap skin necrosis incidence (9.7%). When taking into account the size of the primary defect prompting reconstruction and prior recipient bed irradiation history, our cohort shows reasonable complication rates. Distal tip skin necrosis rate was also low, which suggests that the distal third’s vascular supply should not be a limiting factor. Level of evidence Level IV, therapeutic study.

Optimizing esthetics in delayed breast reconstruction is a challenge, as there is a lack of breast volume and native breast skin. Abdominal flaps are commonly used for patients desiring autologous reconstruction, but the result is an unnatural appearing breast as non-breast skin is used to recreate the breast mound. To improve esthetics for these patients, we propose a two-staged approach utilizing prepectoral tissue expander placement. A retrospective analysis of all patients undergoing delayed breast reconstruction with prepectoral tissue expander placement at our single tertiary-care institution was performed. We collected demographics, complication rates, and surgical information from both the first and second stage surgeries for all patients. Charts were further reviewed to determine whether patients pursued revisions. Five patients (eight breasts) met inclusion criteria. Following tissue expander placement, there was one incidence of seroma (n = 1), and no other complication occurred. Complications after autologous reconstruction included donor-site hematoma (n = 1), recipient-site dehiscence (n = 1), and recipient-site fat necrosis (n = 1). No flap losses or other complications occurred for our series of patients. Delayed, two-staged autologous breast reconstruction requires an additional short, surgical procedure that results in a lifelong improved breast esthetic and better lower-pole ptosis. Complication rates remain low, which makes this a viable and safe option to improve outcomes for patients seeking delayed autologous reconstruction. Our study utilizes prepectoral expander placement, which we believe will further enhance the quality of patient care. Level of evidence: Level V, therapeutic study.

Quality of life (QOL) is an important outcome measure after ventral hernia repair (VHR). The Hernia-Related QOL Survey (HerQLes) is a specific survey tool for QOL after VHR. Studies comparing QOL in patients with biologic mesh repairs (BMRs) and synthetic mesh repairs (SMRs) are lacking.A survey based on the HerQLes was administered via e-mail to 974 patients who had undergone VHR at Stanford Medical Center. From 175 patients who were included in the study, the mean HerQLes scores and postoperative complications were compared between patients with suture repairs (SR), BMR, SMR, with and without component separation, and different types of SMR.Quality of life was lower in patients with hernias of 50 cm or greater, obesity, history of tobacco use, previous abdominal surgeries, hernia recurrences, and postoperative complications (P0.05). Patients with SR and SMR had a comparable QOL (71.58 vs 70.12, P = 0.75). In patients with Modified Ventral Hernia Working Group grade 2 hernias, a significantly lower QOL was found after BMR compared with SMR. Postoperative complications did not significantly differ between the groups. Recurrence rates were comparable between MR (10.4%) and SR (8.3%, P = 0.79), but higher in BMR (21.7%) compared with SMR (6.6%, P0.05).Previous abdominal surgeries, previous hernia repairs, tobacco use, and hernia sizes of 50 cm or greater negatively affect QOL after VHR. Our data indicate that QOL is comparable between patients with SMR and SR, however, is lower in patients with Modified Ventral Hernia Working Group grade 2 and BMR compared with SMR, raising the benefit of BMR in light of its higher cost into question.

The decision to intubate acute burn patients is often based on the presence of classic clinical exam findings. However, these findings may have poor correlation with airway injury and result in unnecessary intubation. We investigated flexible fiberoptic laryngoscopy (FFL) as a means to diagnose upper airway thermal and inhalation injury and guide airway management. A retrospective chart review of all burn patients who underwent FFL from 2013 to 2017 was performed. Their charts were reviewed to determine the indications for FFL including the historical data and physical exam findings that indicated airway injury as well as patient age, TBSA, type and depth of burn injury, carboxyhemoglobin level, and clinical course. Fifty-one patients underwent FFL, with an average TBSA of 6.5% (range 0.5-38.0%) and carboxyhemoglobin level of 3.5%. Burn mechanism was flame (35.3%) or flash (51.0%), with 50% occurring in enclosed spaces. In all cases, the decision to perform FFL was based on physical exam findings meeting criteria for intubation, including facial burns, singed nasal hairs, nasal soot, voice change, throat pain or abnormal sensation, shortness of breath, carbonaceous sputum, wheezing, or stridor. Based on FFL, 9 patients (17.7%) were treated with steroids, 28 patients (54.9%) received supportive care, and 6 patients (11.8%) had repeat FFL for monitoring. One patient was intubated after repeat FFL examination. All patients who underwent FFL met traditional criteria for intubation based on exam, however 98% were monitored without issues based on FFL findings. FFL is a valuable tool that can lead to fewer intubations in acute burn patients with a stable respiratory status for whom history and physical exam suggest upper airway injury.

Background: Zinc performs many biochemical and physiological functions; however, toxicological studies demonstrate that Nano-zinc oxide has harmful effects on human health and environmental species in high concentrations. Objective: The aim of this study was to investigate the toxicity of zinc oxide nanoparticles on reproductive tissues of female rat. Materials and Methods: Eighty female Wistar adult rats weighing 180–200 gr, divided into eight groups (n= 10 in each group) including control, sham (treated with saline), and six groups injected with different doses of zinc oxide nanoparticle with 10–30 nanometer size (4, 8, 25, 50, 100, and 200 mg/kg) twice a week for four weeks. At the end of the study, the rats were bled and slaughtered; the Ovary and Uterus were taken for histopathology studies and blood samples were transferred to the laboratory for biochemical analysis. Results: Microscopic diagnoses in ovary tissue were included; increase in the corpus luteum, follicular cysts, inflammatory cells infiltration and fibrosis. Histopathological changes in ovary in a dose-dependent manner. In uterus tissue the lesions consisted; epithelial destruction, hyperplasia of endometrial glands. The Estrogen and Progesterone level in the serum of rats increased in low doses and reduced in a dose-dependent manner at high doses. Conclusion: The results of the current study proved the toxicity of zinc oxide nanoparticles on the ovary and uterus organs at high concentrations, so further investigation is needed to reduce these effects. Key words: Zinc oxide, Nanoparticles, Ovary, Uterus, Toxicity.

Purpose The present study aims to summarize the current understanding of probable mechanisms and claims of adverse effects of SARS-CoV-2 on male fertility potential. Methods Our search was including original articles, reviews, guidelines, letters to the editor, comments on guidelines, and editorials, regarding the male reproductive system. We used the words SARS-CoV-2, coronavirus, severe acute respiratory syndrome coronavirus 2, “2019 ncov,” testis, sperm, male factor infertility, fertility treatment, semen, assisted reproductive technology (ART), sexual transmission, and ACE2. Results Data showed coronavirus affects men more than women because of more expression of 2019 nCoV receptors (ACE2 and TMPRSS2) in testicular cells. Also, “Bioinformatics Analysis” suggests that sperm production may be damaged, since “Pseudo Time Analysis” has shown disruption in spermatogenesis. “Gene Ontology” (GO) showed an increase in viral reproduction and a decrease in sperm production-related terms. Recently, SARS-COV-2 mRNA and protein were detected in the semen of patients that had recovered from SARS-CoV-2 infection. Therefore, the probable disruption of blood-testis barrier (BTB) in febrile diseases is suspected in the acute phase of the disease enabling viral entry into the testes. Not only is spermatogenesis disturbed, but also disturbs gonadotropin, androgens, and testosterone secretion during SARS-CoV-2 infection. No sexual transmission has been reported yet; however, detection of the virus in semen still makes the sexual transmission an open question. Conclusion There is a concern that male fertility may be disturbed after the SARS-CoV-2 infection. Therefore, follow-up of the reproductive functions and male fertility may be necessary in recovered cases, especially in aged men.

SUMMARY Virtual reality and other technological advancements both inside and outside the operating room have shown an exponential increase in the past two decades. Surgical technique and finesse in delicate procedures have become ever more important, and the onus is on plastic surgeons and plastic surgery residents to meet these needs to provide the best outcomes possible to patients. The ability to learn, simulate, and practice operating in a fashion that poses no harm to any patient is truly a gift from technology to surgery that any surgeon could benefit from, whether trainee or attending. This application of technology and simulation has been demonstrated in other fields such as in the airline industry with flight simulation. The ability to learn, synthesize, and incorporate learned materials and ideas through virtual, augmented, and mixed reality tools offers a great opportunity to put our field at the forefront of a paradigm shift in surgical education. The critical utility of digital education could not be further emphasized any more than in the unfortunate and infrequent situation of a worldwide pandemic. This article reviews some of the important recent technologies that have developed and their applications in plastic surgery education and offers a look into what we can expect in the future.

Patient: Male, 48-year-old Final Diagnosis: Hemophaocytic lymphohistiocytosis Symptoms: Abdominal pain • nausea • vomiting • weight loss Medication:— Clinical Procedure: — Specialty: Rheumatology Objective: Rare disease Background: Sarcoidosis is a systemic inflammatory disorder characterized by a classic pathologic feature of non-caseating granulomas involving any organ system. Hemophagocytic lymphohistiocytosis (HLH) is a catastrophic cytokine surge characterized by dysregulation of the macrophage response, which can be rapidly fatal. Recognition of HLH has been increasing over the past decade. HLH can present with features of sepsis that can make the diagnosis challenging and requires high clinical suspicion. Case Report: We report a case of a 48-year-old African American male with a past medical history of sarcoidosis infiltrating the lymph nodes, liver, and bone marrow with initial presentation of abdominal pain, nausea, vomiting, and weight loss of 100 pounds over 8 months. Sepsis was suspected, but the patient clinically deteriorated with vancomycin and cefepime. Fevers, bone marrow biopsy, anemia, thrombocytopenia, elevated ferritin, and elevated soluble receptor interleukin 2 confirmed HLH. The patient was treated with etoposide and dexamethasone with poor response and died from cardiac arrest. Conclusions: Sarcoidosis associated with HLH is an extremely rare phenomenon with only 10 cases reported in the literature. Early clinical suspicion can be challenging as this condition is a sepsis-mimicker. To reduce mortality, prompt initiation of therapy is a key determinant in patients who are clinically deteriorating despite treatment for sepsis.

BACKGROUND Delayed-immediate breast reconstruction has traditionally involved placement of tissue expanders (TE) in the subpectoral (SP) position. Development of acellular dermal matrices has renewed interest in the prepectoral (PP) pocket, which avoids extensive muscle manipulation. We compare complication rates between PP and SP TE placement in autologous delayed-immediate breast reconstruction. METHODS A retrospective chart review of patients undergoing autologous, delayed-immediate breast reconstruction at our institution (June 2009 to December 2018) was performed. Demographics, comorbidities, perioperative information, and complication incidence ≤12 months' follow-up were collected from first- and second-stage surgeries. Complications were modeled using univariable and multivariable binary logistic regressions. RESULTS A total of 89 patients met the inclusion criteria, and data from 125 breast reconstructions were evaluated. Complication rates following TE placement trended lower in the PP cohort (28.8% vs 37%, P = 0.34). Overall complication rates following autologous reconstruction were significantly lower for PP reconstructions (7.7% vs 23.3%, P = 0.02). Multivariable regression showed TE position (P = 0.01) was a significant predictor of ≥1 complication following autologous reconstruction. Time delay between first- and second-stage surgeries was greater for SP reconstructions (199.7 vs 324.8 days, P < 0.001). Postoperative drains were removed earlier in the PP cohort (8.6 vs 12.0 days, P < 0.001). Mean follow-up time was 331.3 days. CONCLUSIONS Prepectoral reconstruction in the delayed-immediate autologous reconstruction patient leads to significantly lower complication rates, shorter duration between first- and second-stage surgeries, and shorter times before removal of breast drains compared with SP reconstructions.

Patient: Female, 21-year-old Final Diagnosis: Diffuse alveolar hemorrhage Symptoms: Cough • dyspnea • fever • rash • sore throat Medication: — Clinical Procedure: — Specialty: Rheumatology Objective: Rare disease Background: Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by autoantibody production leading to inflammation in multiple organs; it commonly affects young women in their child-bearing years. Clinical manifestations are diverse and range from mild arthritis to diffuse alveolar hemorrhage (DAH). DAH is a rare and devastating complication of SLE that carries a mortality rate of up to 50%, despite aggressive therapy. Case Report: A 21-year-old primigravida at 16 weeks gestation presents with a productive cough, rash, sore throat, and high-grade fever. Chest x-ray suggested multifocal pneumonia. Patient deteriorated despite antibiotics and intravenous (IV) fluids. She developed worsening anemia, leukopenia, and thrombocytopenia. Autoimmune work-up was positive for Coombs, antinuclear antibody, anti-smith antibody, and hypocomplementemia. Skin biopsy was consistent with SLE. SLE vasculitis was suspected. She required mechanical intubation for rapid respiratory deterioration, with CT thorax suggesting ARDS. Bronchoscopy was done and confirmed DAH. Her course was further complicated with retinopathy and acute pancreatitis associated with SLE. She was treated with IV steroids, IV cyclophosphamide, and plasmapheresis, with significant clinical improvement and successful extubation. She delivered a healthy baby at 32 weeks gestation. Conclusions: Early recognition and initiation of treatment is critical to survival in DAH and requires a high index of clinical suspicion. Treatment includes high-dose steroids, cyclophosphamide, and plasma exchange. Pregnancy increases the risk of adverse outcome in SLE. Seven cases of DAH in pregnant patients with SLE have been reported. Here, we report a catastrophic presentation of DAH, acute pancreatitis, and retinopathy in a pregnant patient with newly diagnosed SLE.

Breast implants have been related to breast implant-associated anaplastic large cell lymphoma (BIA-ALCL). While some research has been conducted to study BIA-ALCL incidence, little is known regarding surgeon concern about the disease. This study aims to determine surgeon concern about BIA-ALCL within the European plastic surgeon community as related to their practice of breast plastic surgery. A 27-question online survey was sent to 2353 members of the European Plastic Surgery Society and EURAPS. 240 surgeons responded (10.2%) from 18 different societies. Questions were related to demographics, exposure to BIA-ALCL cases, clinical practices, awareness, and concern. Univariate and multivariable analyses were used. Of surveyed surgeons, 8% had encountered a case of BIA-ALCL, while 73% were concerned about the disease. The rate of concern seemed to be influenced by seven of the variables studied. However, multivariate analysis demonstrated that none of the combined variables analyzed predicted concern or disclosure of the risks of BIA-ALCL to patients. Textured silicone-filled implants were implicated in the disease (56.5% of cases, P = 0.005). Mentor® and Polytech® were the two brands involved in most of the reported cases (20% each). Consistent with epidemiological reports worldwide, this study confirms that BIA-ALCL is more prevalent in patients undergoing placement of textured silicone implants, the use of which was greater among surgeons not concerned about the risks of BIA-ALCL. Surgeons appear to approach their patients with similar risk disclosures regardless of practice pattern and type of breast implant used, but not regardless of their concern about the disease. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

Background: Proper nutrition is critical in healing burn wounds. Tube feeding is routinely employed to help meet nutritional goals; however, the complexities of burn care frequently involve pausing feeds. Volume-based tube feeding delivers a 24-h goal volume compared to administering a fixed hourly rate. This study aims to assess whether volume-based tube feeding is superior to traditional rate-based tube feeding at meeting nutritional goals. Methods: Starting in January 2016, a single burn center implemented a volume-based tube feeding protocol. All patients for the calendar year 2016 who required tube feeds for at least 5 continuous days were compared to all patients from the prior calendar year 2015. Demographics, burn characteristics, and tube feeding details were compared with univariable analysis. Daily tube feed goals were the primary outcome and length of stay was assessed as a secondary outcome; both were evaluated with multivariable analysis using linear modeling. Results: Thirty patients met study criteria. The cohort was 30.0% female and 93.3% were flame burns, with a mean age of 44.2 years and mean total body surface area (TBSA) of 30.6%. All patients suffered 3rd degree burns. Pausing tube feeds for fasting before an operation was the most common reason feeds were held accounting for 33.7% of all held days. Volume-based tube feeding was the only independent predictor of increasing tube feed goals with an average goal met of 91.4% vs. 74.1%, (linear coefficient 10.05, 95% CI 4.49–15.61, p-value 0.001). Tube feed hours held was the only independent predictor of lower tube feed goals (linear coefficient −5.52, 95% CI −7.73–−3.31, p-value

Recent work has established that glutamatergic synaptic activity induces transient sodium elevations in grey matter astrocytes by stimulating glutamate transporter 1 (GLT-1) and glutamate-aspartate transporter (GLAST). Glial sodium transients have diverse functional consequences but are largely unexplored in white matter. Here, we employed ratiometric imaging to analyse sodium signalling in macroglial cells of mouse corpus callosum. Electrical stimulation resulted in robust sodium transients in astrocytes, oligodendrocytes and NG2 glia, which were blocked by tetrodotoxin, demonstrating their dependence on axonal action potentials (APs). Action potential-induced sodium increases were strongly reduced by combined inhibition of ionotropic glutamate receptors and glutamate transporters, indicating that they are related to release of glutamate. While AMPA receptors were involved in sodium influx into all cell types, oligodendrocytes and NG2 glia showed an additional contribution of NMDA receptors. The transporter subtypes GLT-1 and GLAST were detected at the protein level and contributed to glutamate-induced glial sodium signals, indicating that both are functionally relevant for glutamate clearance in corpus callosum. In summary, our results demonstrate that white matter macroglial cells experience sodium influx through ionotropic glutamate receptors and glutamate uptake upon AP generation. Activity-induced glial sodium signalling may thus contribute to the communication between active axons and macroglial cells.

Background: Asthenozoospermia is one of the etiologies for male factor infertility. It was shown that any abnormality in protamines genes, reduction of protamines transcript and protamines deficiency may play a key role in asthenozoospermia. Objective: The aim of the current study was the evaluation of protamine-1 and 2 genes (PRM1 and PRM2) polymorphisms in asthenozoospermic men. Materials and Methods: In this case-control study, the samples were corresponded to asthenozoospermic specimens of infertile men. The normozoospermic samples were considered as the control group. DNA sequence amplification was performed using four PRM1 and PRM2 primers, designed from 5' to 3' flank regions. The human PRM1 and PRM2 gene sequences were screened in search of potential mutations in highly prevalent polymorphism regions in asthenozoospermia versus normozoospermia. Results: Totally, nine highly prevalent polymorphism regions between the forward and reverse primers were screened. Three of them corresponded to PRM1 and six to PRM2. The most prevalent polymorphism regions in PRM1 were related to 102G>T (rs35576928), 49C>T (rs140477029) and 139C>A (rs737008). In the PRM2, 6 highly prevalent polymorphisms regions were screened, including 248C>T (rs779337774), 401G>A (rs545828790), 288C>T (rs115686767), 288G>C (rs201933708), 373C>A (rs2070923), and 298G>C (rs1646022). The allele frequencies of three upper mentioned single nucleotide polymorphisms in asthenozoospermic men including 373C>A, 298G>C and 139C>A was higher than the control group. Conclusion: Our findings indicated that the frequency of some altered genotypes in asthenozospermia was slightly higher than control group. We proposed more extensive studies to be sure that; these genotypes can precisely be related to diagnosis of asthenozoospermia, as the molecular markers.

To have met inclusion criteria, the patient must have undergone delayed breast reconstruction without having had any prior breast reconstruction The transverse rectus abdominis Muscle (TRAM) flap was used in 2 breasts, and the muscle-sparing TRAM flap was used in 6 breasts (Table 1) Dear Sir, We thank Mantelakis et al [[1]] for their interest in our article "Delayed, two-staged autologous breast reconstruction: an approach to improving delayed reconstructive outcomes" [[2]] [Extracted from the article] Copyright of European Journal of Plastic Surgery is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission However, users may print, download, or email articles for individual use This abstract may be abridged No warranty is given about the accuracy of the copy Users should refer to the original published version of the material for the full abstract (Copyright applies to all Abstracts )

Apoptosis of infected host macrophages by Leishmania spp. is mainly addressed as one of the survival mechanisms of the parasite. However, there is no eligible data about whether tumor suppressor p53 could induce the apoptosis of host lymphocytes-treated Leishmania major via the mitochondrial intrinsic pathway. In this study, the amastigotes of L. major obtained from ten cutaneous leishmaniases (CL) patients were separately isolated and cultured in N.N.N and RPMI 1640 media. L. major was definitely confirmed by targeting Cyt b gene following sequencing. Subsequently, 2–3 × 106 lymphocytes obtained from ten healthy individuals were isolated and co-cultured with 1–2 × 106 L. major promastigotes. Following 6 h of exposure time, the enzymatic activity of caspase-3 was determined by fluorometric assay in each L. major-treated lymphocytes and cell control (only lymphocyte). The mRNA expressions of Bax, Bcl-2, p53, and caspase-3 genes were assessed by quantitative real-time-PCR analysis following 6 to 9 h of exposure times. The Bcl-2 mRNA expression in L. major-treated lymphocytes was 100-fold down-regulated relative to cell control. The mRNA expressions of p53 and caspase-3 were over-expressed 1.8- and 3.2-fold up-regulated relative to control lymphocytes, respectively. The Bax/Bcl-2 ratio and caspase-3 activity were higher than the control group (Pv

In brain grey matter, excitatory synaptic transmission activates glutamate uptake into astrocytes, inducing sodium signals which propagate into neighboring astrocytes through gap junctions. These sodium signals have been suggested to serve an important role in neuro-metabolic coupling. So far, it is unknown if astrocytes in white matter-that is in brain regions devoid of synapses-are also able to undergo such intra- and intercellular sodium signalling. In the present study, we have addressed this question by performing quantitative sodium imaging in acute tissue slices of mouse corpus callosum. Focal application of glutamate induced sodium transients in SR101-positive astrocytes. These were largely unaltered in the presence of ionotropic glutamate receptors blockers, but strongly dampened upon pharmacological inhibition of glutamate uptake. Sodium signals induced in individual astrocytes readily spread into neighboring SR101-positive cells with peak amplitudes decaying monoexponentially with distance from the stimulated cell. In addition, spread of sodium was largely unaltered during pharmacological inhibition of purinergic and glutamate receptors, indicating gap junction-mediated, passive diffusion of sodium between astrocytes. Using cell-type-specific, transgenic reporter mice, we found that sodium signals also propagated, albeit less effectively, from astrocytes to neighboring oligodendrocytes and NG2 cells. Again, panglial spread was unaltered with purinergic and glutamate receptors blocked. Taken together, our results demonstrate that activation of sodium-dependent glutamate transporters induces sodium signals in white matter astrocytes, which spread within the astrocyte syncytium. In addition, we found a panglial passage of sodium signals from astrocytes to NG2 cells and oligodendrocytes, indicating functional coupling between these macroglial cells in white matter.

Background Disordered Eating (DE) shows a strong association with athletics and can lead to several negative mental and physical health effects. Traditionally, sports have been grouped based upon whether or not the sport emphasizes leanness as a competing factor. Due to sociocultural factors, risk for DE may also be associated with the sport type. The aim of this review is to critically analyze the available research and data in this field to consider the relationship between DE and sport type to see which factors influence prevalence among athletes Method A systematic review was completed using keywords specific to DE and sport types. Articles were either excluded due to lack of specification of athlete type or failure to use a standardized screening tool or interview for data collection. Results 6 out of 7 studies found a significant increase in DE rates among lean sport types. When classifying by sport type reports were less consistent, but show non-lean sports also have increased rates of DE. Conclusion There are variations in prevalence of DE behaviors depending on athlete type. It is important to identify the risk for DE early in athletes so emphasis can be placed on treatment options to nullify progression to an eating disorder, lower negative impacts on an athlete’s performance, and prevent other negative health effects. Using sport groups is important to clinical practice as well as research, as certain sports may have a higher risk for development of DE.

Aim Scientists have tried to culture and transplant the ovarian tissues (OT), to preserve fertility in cancer patients. However, one of the main limitations to the applicability of this technique is the folliculogenesis disruption after transplantation. Due to the roles exerted by growth differentiation factor-9β (GDF9β), we decided to determine the most effective dose of GDF9β on promotion of folliculogenesis and angiogenesis in sheep OT grafted onto the chick chorioallantoic membrane (CAM). Methods Fresh sheep OT were grafted onto the CAM for 5 days, and divided into four groups based on the addition of increasing doses of GDF9β (0, 150, 200 and 250 ng/mL). Following culture, histological (hematoxylin and eosin [H&E] staining) and immunohistological studies (Ki-67) were done. Fibrotic and necrotic regions were measured using MICROVISIBLE software. For comparing the follicle development rates between the groups as well as differences in the Ki-67-positive follicles, analysis of variance was applied. Results In both 200 and 250 ng/mL GDF9β groups, significantly higher rates of intermediary and primary follicles were observed, also the numbers of good quality follicles increased in the aforementioned groups and the rates of fibrotic and necrotic areas decreased. Moreover, in the 200 and 250 ng/mL GDF9β groups, the number of capillaries and the proliferative activity increased. The lower dose of GDF9β (150 ng/mL) neither activated the primordial follicles nor lead to an increase in the number of growing follicles. Conclusion Addition of high dosages of GDF9β to the OT, grafted onto the CAM resulted in higher folliculogenesis and better transplantation features due to improvement in angiogenesis.

One of the promising approaches for the treatment of cardiac disease is stem cell therapy. In this study, we compared the cardiomyogenic differentiation rate, from human adipose-derived stem cells (hADSCs) in a three-dimensional (3D) hanging drop (HD) spheroid culture system, versus a two-dimensional (2D) culture condition at different concentrations of 5-azacytidine (5-Aza). 5-Azaytidine (5-Aza) is a pyrimidine nucleoside analogue of cytidine that initiates cell differentiation programs through DNA demethylation. The hADSCs were isolated and cultured both in 2D and 3D HD conditions, with either 10 or 50 μM concentrations of 5-Aza. Then DNA content, gene expression, and protein content were analyzed. 3D HD culture resulted in a higher percentage of cells in G0/G1 and S phase in the cell division cycle, whereas 2D culture led to a greater percentage of cells in the G2/M phase. A significantly higher gene expression rate of HAND1, HAND2, cTnI, Cx43, βMHC, GATA4, NKX2.5, and MLC2V was observed in HD treated with 50 μM 5-Aza. This was confirmed by immunocytochemistry. These findings suggest that 50 μM concentration of 5-Aza can induce hADSCs to differentiate into cardiomyocytes. The differentiation rate was significantly higher when accompanied by the 3D HD culture system. This work provides a new culture system for cell differentiation for cardiovascular tissue engineering.

Spirocerca lupi is an international nematode of Canine and wild carnivores. The generic clinical symptoms are regurgitation, vomiting, and pursiness. The parasite usually procreates nodular masses in the esophagus and pectoral aorta. On July 2014, a four-month male sandy fox was found accidentally killed on the road in Bafgh City, Southeast of Yazd, Yazd Province, Iran during necropsy, referring to esophagus, parasites were collected from esophagus and transmissive up to 70% of alcohol. Parasites were recognized as S. lupi. After clarification of lactophenol and staining, the nematodes were recognized as S. lupi according to key guidelines of Taylor. Besides, the tissue samples were isolated from esophagus wall - the appendix site of nematode for histopathological investigation - so they were transmissive up to 10% of buffered formalin and stained with haematoxylin and eosin. Microscopic diagnoses in esophagus were included epithelial destruction, wall thickening, inflammatory cells infiltration, necrosis and vascular thrombosis. With the author’s knowledge, this is the first report and histopathological investigation of S. lupi in sandy fox.

Perivascular endfeet of astrocytes are highly polarized compartments that ensheath blood vessels and contribute to the blood-brain barrier. They experience calcium transients with neuronal activity, a phenomenon involved in neurovascular coupling. Endfeet also mediate the uptake of glucose from the blood, a process stimulated in active brain regions. Here, we demonstrate in mouse hippocampal tissue slices that endfeet undergo sodium signaling upon stimulation of glutamatergic synaptic activity. Glutamate-induced endfeet sodium transients were diminished by TFB-TBOA, suggesting that they were generated by sodium-dependent glutamate uptake. With local agonist application, they could be restricted to endfeet and immunohistochemical analysis revealed prominent expression of glutamate transporters GLAST and GLT-1 localized towards the neuropil vs. the vascular side of endfeet. Endfeet sodium signals spread at an apparent maximum velocity of ∼120 µm/s and directly propagated from stimulated into neighboring endfeet; this spread was omitted in Cx30/Cx43 double-deficient mice. Sodium transients resulted in elevation of intracellular magnesium, indicating a decrease in intracellular ATP. In summary, our results establish that excitatory synaptic activity and stimulation of glutamate uptake in astrocytes trigger transient sodium increases in perivascular endfeet which rapidly spread through gap junctions into neighboring endfeet and cause a reduction of intracellular ATP. The newly discovered endfeet sodium signaling thereby represents a fast, long-lived and inter-cellularly acting indicator of synaptic activity at the blood-brain barrier, which likely constitutes an important component of neuro-metabolic coupling in the brain. GLIA 2017;65:293-308.

Besides astrocytes and oligodendrocytes, NG2 proteoglycan-expressing cells (NG2 glia) represent a third subtype of macroglia in the brain. Originally described as oligodendrocyte precursor cells, they feature several characteristics not expected from mere progenitor cells, including synaptic connections with neurons. There is accumulating evidence that the properties of NG2 glia differ between different brain regions and developmental stages. To further analyze this proposed heterogeneity, we studied electrophysiological properties, transcript and protein expression, distribution and proliferative capacity of NG2 glia during postnatal development, focusing on the hippocampus and corpus callosum. All NG2 glia displayed a 'complex' current pattern consisting of voltage- and time-dependent in- and outward currents. In juvenile mice, Kir current densities and rectification index were highly variable and on average significantly lower than in adult animals. Single cell RT-PCR analyses of electrophysiologically characterized cells demonstrated that different glial genes were expressed at variable extent, independent of developmental stage and genetic background. In the hippocampus proper and the corpus callosum, the density of NG2 glia was highest at postnatal days (P)10-12, decreased by ~50 % at P25-35 and then remained stable in adults (P80-90). Interestingly, co-expression of NG2 and S100β, a marker for mature astrocytes, increased from 7 % at P10-12 to 27 % at P25-35 in the hippocampus proper, and then dropped again in the stratum radiatum at P80-90. In the dentate gyrus and corpus callosum, co-expression of NG2 and S100β was very low (3 %) and constant throughout development. Age-related differences were also observed with Ki-67, a proliferation marker. In NG2 glia of the CA1 region, its expression decreased from 16 % at P10-12 to 9 % (P25-35) and then 3 % (P80-90). Triple-stainings revealed that Ki-67 was also expressed in 2-3 % of NG2/S100β-positive cells in the juvenile and mature stratum radiatum, indicating that the latter, in contrast to S100β-positive astrocytes, still host proliferative potential. Taken together, we found significant differences in transcript and protein expression, electrophysiological properties and proliferative capacity of NG2 glia in the mouse forebrain, suggesting the co-existence of several subpopulations of NG2 glia. Our data thus support the idea of a substantial regional and developmental heterogeneity in this subtype of macroglia.

Objective: In early gestation, fetal wounds heal without fibrosis in a process resembling regeneration. Elucidating this remarkable mechanism can result in tremendous benefits to prevent scarring. Fetal mouse cutaneous wounds before embryonic day (E)18 heal without scar. Herein, we analyze expression profiles of fetal and postnatal wounds utilizing updated gene annotations and pathway analysis to further delineate between repair and regeneration.

Background: Fetal bovine serum (FBS) is widely used in cell culture laboratories, risk of zoonotic infections and allergic side effects create obstacles for its use in clinical trials. Therefore, an alternative supplement with proper inherent growth-promoting activities is demanded. Objective: To find FBS substitute, we tested human umbilical cord blood serum (hUCS) for proliferation of human umbilical cord matrix derived mesenchymal stem cells (hUC-MSCs) and human bone marrow-derived mesenchymal cells (hBM-MSCs). Materials and Methods: Umbilical cord blood of healthy neonates, delivered by Caesarian section, was collected and the serum was separated. hUC-MSCs and hBM-MSCs were isolated and characterized by assessment of cell surface antigens by flow cytometry, alkaline phosphatase activity and osteogenic/adipogenic differentiation potential. The cells were then cultured in Iscove's Modified Dulbecco's Medium (IMDM) by conventional methods in three preparations: 1- with hUCS, 2- with FBS, and 3- without serum supplements. Cell proliferation was measured using WST-1 assay, and cell viability was assessed by trypan blue staining. Results: The cells cultured in hUCS and FBS exhibited similar morphology and mesenchymal stem cells properties. WST-1 proliferation assay data showed no significant difference between the proliferation rate of either cells following hUCS and FBS supplementation. Trypan blue exclusion dye test also revealed no significant difference for viability between hUCS and FBS groups. A significant difference was detected between the proliferation rate of stem cells cultured in serum-supplemented medium compared with serum-free medium. Conclusion: Our results indicate that human umbilical cord serum can effectively support proliferation of hBM-MSCS and hUC-MSCs in vitro and can be used as an appropriate substitute for FBS, especially in clinical studies.

Summary Background Autologous breast reconstruction is considered by many to be the gold standard reconstructive modality following mastectomy. Despite the advantages of autologous reconstruction, however, surgeons have been cautious in recommending this approach to patients who desire to become pregnant postoperatively because of concerns related to abdominal wall morbidity. While intuitive, this approach does not appear to be based on robust data. Hence, the authors examined the clinical outcome in patients who became pregnant following autologous breast reconstruction. Methods Patients who underwent autologous breast reconstruction with free abdominal flaps that required an incision in the anterior rectus sheath were identified. Of them, patients who became pregnant post reconstruction were included for subsequent analysis. Of particular interest were any peripartal and postpartal complications that could be attributed to the preceding abdominal flap harvest. Additionally, a systematic review of the literature was performed. Results We identified five patients who met inclusion criteria. All five patients underwent bilateral breast reconstruction with free muscle-sparing transverse rectus abdominis musculocutaneous (MS-TRAM) flaps. None of the patients had any preexisting abdominal wall morbidity. All five patients proceeded to full-term pregnancy and successfully delivered newborns, four of which were delivered by normal vaginal delivery and one by cesarean section. No abdominal wall complications were noted during pregnancy, delivery, or postpartum. Conclusion Contemporary data do not support the notion that breast reconstruction with free abdominal flaps is contraindicated in the setting of desired or planned pregnancy.

Context: Rosa damascena L. (Rosaceae) (RD) essential oil and extracts are commonly used as a flavour in herbal medicine which increase libido. Previous studies have shown inhalation of RD flower’s oil increases libido and causes protective effects in formaldehyde (FA)-induced testicular damage. Objective: The protective effects of aqueous extract of RD on the male reproductive system of mice were examined following FA-induced damage. Materials and methods: Forty-eight adult NMRI male mice were randomly assigned to six groups (n = 8): control (normal saline, 10 mg/kg); RD40 (40 mg/kg, p.o.); FA treated (10 mg/kg of 10%, i.p.) and FA + RD treated at 10, 20 and 40 mg/kg (FA + RD10), (FA + RD20) and (FA + RD40), respectively, for 40 days. At the end of treatment regimes, serum testosterone (T) level and the reproductive activity, viz. body/organ weights, testicular structure and sperm characteristics were studied. Results: Formaldehyde administration significantly decreased serum T level (p

Purpose: Since its discovery in December 2019, the novel coronavirus SARS-CoV-2 has spread globally, causing the current COVID-19 (coronavirus disease-19) pandemic. As there is an increase of infections in the male population, concerns have emerged about the potential impact of COVID-19 on male reproductive organs and male fertility. Therefore, this study systematically investigates the current evidence of SARS-CoV-2 impact on male reproduction and pregnancy outcomes, discussing them in light of the evidence published on other coronaviruses. Materials and Methods: Literature search was carried out according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. A total of 24 original articles were included for the analysis, investigating the effects of the infection on semen parameters, male reproductive hormones, and pregnancy outcomes. Further, a Strengths-Weaknesses- Opportunities-Threats (SWOT) analysis was conducted based on the available evidence linking the virus with male reproduction and conception. Results: Although there is limited data, viral mRNA has been identified in semen of infected men, with some evidence of altered seminal parameters. Low testosterone and dihydrotestosterone with raised luteinizing hormone has been reported as well as preterm delivery in pregnant women; however, data regarding vertical transmission remains contradictory and inconclusive. Conclusions: The recent literature provides evidence that male gonads may be potentially vulnerable to SARS-CoV-2 infection, recommending caution to pregnant women and couples planning natural pregnancy or assisted reproduction.

Background The human genome contains variants ranging in size from small single nucleotide polymorphisms (SNPs) to large structural variants (SVs). High-quality benchmark small variant calls for the pilot National Institute of Standards and Technology (NIST) Reference Material (NA12878) have been developed by the Genome in a Bottle Consortium, but no similar high-quality benchmark SV calls exist for this genome. Since SV callers output highly discordant results, we developed methods to combine multiple forms of evidence from multiple sequencing technologies to classify candidate SVs into likely true or false positives. Our method (svclassify) calculates annotations from one or more aligned bam files from many high-throughput sequencing technologies, and then builds a one-class model using these annotations to classify candidate SVs as likely true or false positives. Results We first used pedigree analysis to develop a set of high-confidence breakpoint-resolved large deletions. We then used svclassify to cluster and classify these deletions as well as a set of high-confidence deletions from the 1000 Genomes Project and a set of breakpoint-resolved complex insertions from Spiral Genetics. We find that likely SVs cluster separately from likely non-SVs based on our annotations, and that the SVs cluster into different types of deletions. We then developed a supervised one-class classification method that uses a training set of random non-SV regions to determine whether candidate SVs have abnormal annotations different from most of the genome. To test this classification method, we use our pedigree-based breakpoint-resolved SVs, SVs validated by the 1000 Genomes Project, and assembly-based breakpoint-resolved insertions, along with semi-automated visualization using svviz. Conclusions We find that candidate SVs with high scores from multiple technologies have high concordance with PCR validation and an orthogonal consensus method MetaSV (99.7 % concordant), and candidate SVs with low scores are questionable. We distribute a set of 2676 high-confidence deletions and 68 high-confidence insertions with high svclassify scores from these call sets for benchmarking SV callers. We expect these methods to be particularly useful for establishing high-confidence SV calls for benchmark samples that have been characterized by multiple technologies. Electronic supplementary material The online version of this article (doi:10.1186/s12864-016-2366-2) contains supplementary material, which is available to authorized users.