Canine testes thin sections culture

Canine testes thin section culture Testes tissue culture systems would provide a tool to elucidate spermatogenesis mechanisms, with the aim of genetic preservation of mammals, especially endangered species. Our experiment aims to develop a culture system capable of producing viable mammalian sperm cells in vitro. Dogs were chosen as the model organism because testes are readily available. Canine testes were obtained from a local veterinary clinic. Thin sections were generated using a commercial electric slicer. They then were cleaned using Dulbecco’s Phosphate-Buffered Saline (DPBS) supplemented with antibiotics then cultured in a modified Tissue Culture Medium 199 (TCM-199). Sections were cultured in an environment aimed to best reflect realistic physiological conditions, that is 7%CO2 : 7%O2 : balanced N2 at 37oC. Finally, the sections were stained with live/dead cell stain and observed under a fluorescence microscope to determine viability. Numerous live stained nuclei were observed, proving their high viability after 21 days of culturing. Sections reformed during culture to assume a tiny testes-like morphology. Fungal contamination was detected in all culture dishes at various time points during the experiment from unknown sources. The sections then were washed with DPBS supplemented with antimycotic before being again cultured in fresh medium. For ongoing experiments, the culture system will be revised to prevent fungal contamination. While spermatogenesis takes approximately 60 days in vivo, testes thin sections were maintained for 21 days, therefore culture duration will be extended in the future. Overall, our result demonstrated a cost-effective culture system to potentially obtain viable mammalian sperm cells

Canine testes thin sections culture​

Canine testes thin section culture Testes tissue culture systems would provide a tool to elucidate spermatogenesis mechanisms, with the aim of genetic preservation of mammals, especially endangered species. Our experiment aims to develop a culture system capable of producing viable mammalian sperm cells in vitro. Dogs were chosen as the model organism because testes are readily available. Canine testes were obtained from a local veterinary clinic. Thin sections were generated using a commercial electric slicer. They then were cleaned using Dulbecco’s Phosphate-Buffered Saline (DPBS) supplemented with antibiotics then cultured in a modified Tissue Culture Medium 199 (TCM-199). Sections were cultured in an environment aimed to best reflect realistic physiological conditions, that is 7%CO2 : 7%O2 : balanced N2 at 37oC. Finally, the sections were stained with live/dead cell stain and observed under a fluorescence microscope to determine viability. Numerous live stained nuclei were observed, proving their high viability after 21 days of culturing. Sections reformed during culture to assume a tiny testes-like morphology. Fungal contamination was detected in all culture dishes at various time points during the experiment from unknown sources. The sections then were washed with DPBS supplemented with antimycotic before being again cultured in fresh medium. For ongoing experiments, the culture system will be revised to prevent fungal contamination. While spermatogenesis takes approximately 60 days in vivo, testes thin sections were maintained for 21 days, therefore culture duration will be extended in the future. Overall, our result demonstrated a cost-effective culture system to potentially obtain viable mammalian sperm cells

Leader-Follower Model and Impact of Mobility on Consensus Building

Wireless sensor networks are an indispensable tool in this highly connected world. WSNs have been the focus of research efforts in areas of communication, electronics and control for many years. Advancements in the fields of MEMS, RF and digital circuit technology has led to the development of low cost and extremely power efficient smart sensors. This has led to the need of a fast, reliable and inexpensive method of consensus building for these sensor networks. Basic concepts of graph theory and consensus building are explained in this thesis. This thesis reviews the models and strategies for consensus building present in the literature. The shortcomings of these models are explained through examples and a leader-follower model based consensus building strategy is presented. Algorithm to convert any graph into a bipartite graph by edge removal and a strategy to select effective leaders based on a weighted combination of node centrality, ratio of leaders to the total number of nodes and presence of leaf nodes in the group is presented in this thesis. Proposed leader-follower model is compared against classic models for consensus building are compared and proven to be better. Mobility is studied using deterministic and random mobility models to show the improvement in convergence rate of the network. It is shown that mobility can turn any disconnected network into a connected network, which is able to reach consensus

Comparison of the effect of different medicaments on surface reproduction of two commercially available Polyvinyl Siloxane impression materials: an Invitro Study

Objective:To determine the effect of different retraction cord medicaments on surface detail reproduction of polyvinyl siloxane impression materials and compare this effect on any two brands of commercially available polyvinyl siloxane impression materials. Material and methods: Four stainless steel dies were made according to ADA specification no.19. Three dies were treated with aluminium chloride (5%), ferric sulphate (13.3%) and epinephrine (0.1%) while the fourth one was left untreated to serve as control. Two impression materials (Dentsply and 3M ESPE) were used. Results: All the three medicaments adversely affected the surface detail reproduction of both the brands of the polyvinyl siloxane impression materials. These effects were statistically significant as compared to untreated control. The impressions of 3M ESPE brand have shown better surface detail reproduction as compared to Dentsply impression material. Conclusion: Surface detail reproduction of the polyvinyl siloxane impression materials is adversely affected by the retraction cord medicaments. The presence of moisture or any traces of the medicaments should be removed from the tooth surface to provide a dry field for the correct reproduction of the surface detail of these material

Viral Hepatitis in COVID-Era

The novel SARS-CoV-2 which originated in Wuhan and has spread worldwide is well known to cause severe acute respiratory syndrome. The most common clinical presentations that patients exhibited are fevers, shortness of breath, cough, diarrhea, loss of smell/taste. However, this virus has also been observed to cause multi-organ failure. We present the case of a 44yo F with a PMHx of GERD, Hashimoto’s thyroiditis, obesity, HTN, and neuropathy who presented to the hospital with the chief complaint of worsening shortness of breath. The patient was saturating 92% on room air and with ambulation her pulse oximetry dropped to 80%. Remaining vital signs were stable. Chest x-ray displayed patchy bilateral interstitial and alveolar airspace opacities. Prior to coming to the hospital, the patient had indeed visited an urgent care center and tested positive for COVID-19. On arrival, significant labs included ALT/AST 588/321, NA 133, Ferritin 712, CRP 6.3, LDH 284, WBC 3.4, absolute lymphocytes 0.60, and D-dimer 1.27. It was determined to admit the patient for acute hypoxic respiratory failure secondary to covid-19 infection. GI was consulted for the elevated liver enzymes. The patient had no prior history of hepatitis, blood transfusions, recent travel, diarrhea, alcohol abuse, IV drug use or family history of liver disease. She also had no recent medication changes or over the counter supplemental use. Extensive work-up was initiated for the elevated transaminases and results revealed a negative acute hepatitis panel, ANA, autoimmune liver panel, and no evidence of thrombosis. Given the lack of explanation for the patient’s continually rising liver enzymes, it was decided that a liver biopsy would be performed to be able to provide a valid cause for the elevated enzymes. During the patient’s hospitalization, she was placed on high dose steroids for her acute respiratory failure due to the COVID infection. The liver pathology report results were received after 4 days and reported that the liver lobular parenchyma demonstrated mild lobular inflammation with occasional hepatocyte drop-out and mild sinusoidal congestion without steatosis, granuloma or necrosis. These findings may be due to the patient\u27s SARS-CoV-2 infection. This virus has astonished the medical world and there is a great deal of medical knowledge that still needs to be learned about SARS-COV-2. Patients present with a spectrum of clinical presentations ranging from asymptomatic patients while others can develop grave complications such meningoencephalitis. In patients with elevated transaminases, it is essential to rule out acute hepatitis, autoimmune hepatitis, drug induced liver injury and thrombosis prior to proceeding with a liver biopsy to rule out other causes. In this patient, we had ruled out the above and the only explanation for the acute liver injury was in fact COVID-19.https://scholarlycommons.henryford.com/merf2020caserpt/1116/thumbnail.jp

Developing Cryopreservation Methods of Wheat Roots

In the midst of record breaking rates of plant species extinction due to climate change and fungal diseases, a universal cryopreservation method would provide a means for preservation of these many different species. The concept of plant root cryopreservation first emerged in the late 1960’s, and with it came new avenues of preserving tissue for the purposes of agriculture and research. Frozen tissues can be transported and stored more reliably than other more conventional means. When thawed, they have the potential to be cultured and grown. Several different methods of cryopreservation exist. This experiment used the Fast (3°C/minute) and Slow freeze (0.3°C/minute) method with a controlled freezing unit on a wheat plant species. Cryoprotectant solutions containing 10% DMSO or 10% glycerol were used in conjunction with high (3.11 M) and low (1.5 M) sorbitol concentrations. A no sorbitol group was also tested. After being frozen, samples were transferred to a liquid nitrogen tank for storage, and later thawed. Thawed samples were stained with fluorescent dyes to observe live and dead cells under fluorescent microscopy. Root tips were deemed “surviving” if multiple live nuclei were present. In the Glycerol group, Slow freezing showed a higher survival rate, while the DMSO group had good survival rates for both freezing rates. The Slow freeze rate showed greater survival rates overall compared to the Fast rate. Future experiments to develop a universal method will include testing on various plant species, such as potato plants, and succulents

Administration of S-nitrosoglutathione after traumatic brain injury protects the neurovascular unit and reduces secondary injury in a rat model of controlled cortical impact

<p>Abstract</p> <p>Background</p> <p>Traumatic brain injury (TBI) is a major cause of preventable death and serious morbidity in young adults. This complex pathological condition is characterized by significant blood brain barrier (BBB) leakage that stems from cerebral ischemia, inflammation, and redox imbalances in the traumatic penumbra of the injured brain. Once trauma has occurred, combating these exacerbations is the keystone of an effective TBI therapy. Following other brain injuries, nitric oxide modulators such as S-nitrosoglutathione (GSNO) maintain not only redox balance but also inhibit the mechanisms of secondary injury. Therefore, we tested whether GSNO shows efficacy in a rat model of experimental TBI.</p> <p>Methods</p> <p>TBI was induced by controlled cortical impact (CCI) in adult male rats. GSNO (50 μg/kg body weight) was administered at two hours after CCI. GSNO-treated injured animals (CCI+GSNO group) were compared with vehicle-treated injured animals (CCI+VEH group) in terms of tissue morphology, BBB leakage, edema, inflammation, cell death, and neurological deficit.</p> <p>Results</p> <p>Treatment of the TBI animals with GSNO reduced BBB disruption as evidenced by decreased Evan's blue extravasation across brain, infiltration/activation of macrophages (ED1 positive cells), and reduced expression of ICAM-1 and MMP-9. The GSNO treatment also restored CCI-mediated reduced expression of BBB integrity proteins ZO-1 and occludin. GSNO-mediated improvements in tissue histology shown by reduction of lesion size and decreased loss of both myelin (measured by LFB staining) and neurons (assayed by TUNEL) further support the efficacy of GSNO therapy. GSNO-mediated reduced expression of iNOS in macrophages as well as decreased neuronal cell death may be responsible for the histological improvement and reduced exacerbations. In addition to these biochemical and histological improvements, GSNO-treated injured animals recovered neurobehavioral functions as evaluated by the rotarod task and neurological score measurements.</p> <p>Conclusion</p> <p>GSNO is a promising candidate to be evaluated in humans after brain trauma because it not only protects the traumatic penumbra from secondary injury and improves overall tissue structure but also maintains the integrity of BBB and reduces neurologic deficits following CCI in a rat model of experimental TBI.</p