Enhancing the quality of inferior oocytes of buffalo for in vitro embryo production: The impact of melatonin on maturation, SCNT, and epigenetic modifications.

Success of animal cloning is limited by oocyte quality, which is closely linked to reprogramming ability. The number of layers of cumulus cells is typically used to assess the quality of oocyte; a minimum of one-third of collected cumulus-oocyte complexes (COCs) are discarded as inferior oocytes because they have less cumulus cells. Melatonin, which has been recognised for its ability to sequester free radicals and perform multiple functions, has emerged as a potentially effective candidate for enhancing inferior oocytes quality and, consequently, embryo development competency. The current study investigates to improve the quality of inferior oocytes by supplementation of melatonin (10-9 M) during in vitro maturation (IVM) and subsequent cloned embryo production and its mechanism. The results indicate that melatonin supplementation significantly (p<0.05) enhances inferior oocytes maturation, reduces oxidative stress by reducing ROS levels, and improves mitochondrial function by boosting GSH levels. The melatonin treatment (10-9 M) enhances the expression of SOD, GPx1, GDF 9, BMP 15, ATPase 6, and ATPase 8 in inferior oocytes. Furthermore, melatonin treatment increases the total cell number in the treated groups, promoting cloned blastocyst formation rates derived from inferior oocytes. Furthermore, compared to the control, 10-9 M melatonin supplementation enhances H3K9ac acetylation and lowers H3K27me3 methylation in cloned blastocysts derived from inferior oocytes. In conclusion, 10-9 M melatonin supplementation during IVM increased inferior oocyte maturation and promoted cloned buffalo embryo development by lowering oxidative stress and promoting epigenetic alterations. These studies show that melatonin may improve the quality of poor oocytes and buffalo cloning.

Crystallizing galactocele: A rare case report and review of literature.

ABSTRACT: Crystallizing galactocele is an uncommon condition that produces a viscous, chalky substance on fine needle aspiration cytology. (FNAC). Both the diagnosis and the management of this illness include the use of FNAC. Here, we discuss the case of a 25-year-old nursing woman who experienced left breast edema lump for two years. The upper outer quadrant of the leftt breast was involved by the hard, small, non-tender, and movable enlargement. The lesion's FNAC produced a thick, milky, and chalky substance. Numerous semi-transparent crystals of various sizes and shapes with angulated edges could be seen in cytological smears against a background of granular and amorphous proteinaceous material. A diagnosis of crystallizing galactocele was made on the basis of the patient's clinical history of lactation and characteristic cytological findings. Due to the rarity of this condition-to the best of our knowledge, less than ten cases of crystallizing galactocele have been documented in medical literature.

Optimising Electroporation Condition for CRISPR/Cas-Mediated Knockout in Zona-Intact Buffalo Zygotes.

Somatic cell nuclear transfer or cytoplasm microinjection has widely been used to produce genome-edited farm animals; however, these methods have several drawbacks which reduce their efficiency. In the present study, we describe an easy adaptable approach for the introduction of mutations using CRISPR-Cas9 electroporation of zygote (CRISPR-EP) in buffalo. The goal of the study was to determine the optimal conditions for an experimental method in which the CRISPR/Cas9 system is introduced into in vitro-produced buffalo zygotes by electroporation. Electroporation was performed using different combinations of voltage, pulse and time, and we observed that the electroporation in buffalo zygote at 20 V/mm, 5 pulses, 3 msec at 10 h post insemination (hpi) resulted in increased membrane permeability and higher knockout efficiency without altering embryonic developmental potential. Using the above parameters, we targeted buffalo POU5F1 gene as a proof of concept and found no variations in embryonic developmental competence at cleavage or blastocyst formation rate between control, POU5F1-KO, and electroporated control (EC) embryos. To elucidate the effect of POU5F1-KO on other pluripotent genes, we determined the relative expression of SOX2, NANOG, and GATA2 in the control (POU5F1 intact) and POU5F1-KO-confirmed blastocyst. POU5F1-KO significantly (p ≤ 0.05) altered the expression of SOX2, NANOG, and GATA2 in blastocyst stage embryos. In conclusion, we standardized an easy and straightforward protocol CRISPR-EP method that could be served as a useful method for studying the functional genomics of buffalo embryos.

Disseminated violaceous plaques in a HIV-1-positive patient from eastern India: A manifestation of Kaposi's sarcoma.

Kaposi's sarcoma (KS) is an angioproliferative disorder primarily of viral etiology, though multiple cofactors are also responsible. Human herpes virus-8, a gamma herpes virus, is considered to be the causative agent. Acquired immunodeficiency syndrome-associated KS has different clinical pictures than those seen in other types of KS. As it progresses rapidly, early institution of highly active antiretroviral therapy (HAART) after proper diagnosis is expected. Though HAART has reduced the prevalence of KS in HIV disease, HAART has not eliminated the disease. Here, we report a case who is HIV 1 and hepatitis B surface antigen positive with numerous violaceous plaques over the face, upper extremities, and trunk along with oral mucosal involvement. He had received ten sessions of electron beam radiotherapy on the face, and the facial lesions have healed with residual hyperpigmentation.

Internal hemipelvectomy with ischiectomy, partial acetabulectomy, and femoral head and neck excision in a cat with a pelvic osteochondroma.

CASE DESCRIPTION: A 2-year-old 5.1-kg (11.2-lb) castrated male Siberian cat was examined because of a history of an abnormal right pelvic limb gait and a 4- to 5-month history of progressive constipation. Radiographs obtained by the referring veterinarian showed an osteoproductive and osteolytic bony lesion that involved the right ischium and filled the obturator foramen. CLINICAL FINDINGS: A hard mass was palpable in the right inguinal area, and rectal examination revealed a smooth bony mass on the ventral aspect of the right pelvic floor with marked reduction in the pelvic canal space. A 3.9 × 3 × 4.6-cm, mineralized mass bridging the right obturator foramen was present on CT images. The ventral component of the mass was slightly larger than its dorsal component, and lysis of the right pubic bone was present. There was no obvious soft tissue involvement. TREATMENT AND OUTCOME: A limb salvage procedure involving internal hemipelvectomy with ipsilateral ischiectomy, contralateral partial ischiectomy, ipsilateral partial acetabulectomy, and femoral head and neck excision was performed. Histologic examination revealed that the mass was an osteochondroma. The cat recovered well and had good functional limb use immediately after surgery. The cat was still alive 1 year after surgery with good limb use. CLINICAL RELEVANCE: Internal hemipelvectomy involving ischiectomy, partial acetabulectomy, and femoral head and neck excision can result in a good functional outcome in cats if the procedure is planned appropriately with a full understanding of the regional anatomy and adherence to surgical oncologic principles.

Moxifloxacin-induced oral erythema multiforme: An unusual adverse effect hitherto unreported.

Moxifloxacin is a fluoroquinolone with excellent activity in community-acquired respiratory tract infections. Common adverse effects are gastrointestinal symptoms, headache, dizziness, etc., Some serious adverse effects include tendon rupture, rhabdomyolysis, peripheral neuropathy, and interstitial nephritis. Cutaneous adverse effects include allergic reactions, angioedema, Steven-Johnson syndrome, and toxic epidermal necrosis. Erythema multiforme (EM), an acute self-limiting disease, most commonly occurs due to infection and rarely due to drugs or systemic disease. EM is classified into EM major and minor, both having skin lesions. A third category of EM has also been described with only oral involvement and without any skin lesions. Oral EM itself is an uncommon entity which has been reported due to nonsteroidal anti-inflammatory drugs. Here, we are reporting a case of moxifloxacin-induced oral EM. After extensive search in PubMed-Medline database, we could not find any such co-occurrence of moxifloxacin-induced oral EM. To the best of our knowledge, this is the first reported case.

Survival time of juvenile dogs with oral squamous cell carcinoma treated with surgery alone: A Veterinary Society of Surgical Oncology retrospective study.

OBJECTIVE: To report the signalment, staging, surgical treatment, and survival time of juvenile dogs treated surgically for oral squamous cell carcinoma (OSCC). STUDY DESIGN: Retrospective study. ANIMALS OR SAMPLE POPULATION: Twenty-five dogs, <2 years of age with OSCC treated with surgery. METHODS: Cases were solicited from the Veterinary Society of Surgical Oncology. Data retrieved included sex, breed, age, weight, clinical signs, tumor location, preoperative diagnostics and staging, histopathological diagnosis with margin evaluation, disease-free interval, and date and cause of death. A minimum follow-up time of 3 months was required for inclusion. RESULTS: Eighteen dogs were <12 months of age, and seven were <24 months. Various breeds were represented, with a mean body weight of 22.3 ± 14.4 kg. No dogs had evidence of metastatic disease prior to surgery. All dogs underwent partial maxillectomy or mandibulectomy. Histological margins were complete in 24 dogs and incomplete in one. No dogs had evidence of metastatic disease or tumor recurrence. The median follow-up time was 1556 days (92 to 4234 days). All dogs were alive at the last follow-up except for one documented death, due to dilated cardiomyopathy. Median disease-specific survival time was not reached. CONCLUSION: The prognosis after wide surgical excision of OSCC in juvenile dogs was excellent. CLINICAL SIGNIFICANCE: OSCC in juvenile dogs can be effectively treated with surgery alone.

Cutaneous manifestations and dermatological sequelae of Covid-19 infection compared to those from other viruses.

In the last few months, there have been numerous reports describing a variety of cutaneous signs associated with COVID-19. Clinicians from Italy were the first to describe the cutaneous manifestations of COVID-19, which were later observed in other parts of the globe. In some cases, cutaneous signs were the only manifestation of COVID-19 rather than the typical syndrome of fever and upper respiratory tract symptoms. However, there is considerable heterogeneity amongst the cutaneous signs described so far, which has been published extensively. Our aim is to summarise the latest studies that have reported the early and late cutaneous signs of COVID-19 and compare them to the most common established viral exanthems.

Psychotic Symptoms in Heroin Withdrawal: A Case Report.

Opiate withdrawal-induced severe exacerbation of psychosis after the sudden withdrawal of an opiate is a known yet uncommon clinical manifestation. We present the case of opiate withdrawal-induced psychosis in a 25-year-old married male patient without any prior psychiatric history of illness, family history, or past hospitalization records. The patient presented with psychotic symptoms such as irritability, delusions of parasitosis, and auditory and visual hallucinations at the time of hospitalization. Symptoms were in a fluctuating course and were not constant throughout the day. Through this case report, our aim is to present a rare instance of heroin-withdrawal-induced psychosis and its successful treatment with antipsychotics.

Preoperative autologous blood donation and transfusion in dogs undergoing elective surgical oncology procedures with high risk of hemorrhage.

OBJECTIVE: To describe preoperative autologous blood donation (PABD) and transfusion in dogs undergoing elective surgical oncology procedures with a high risk of intraoperative hemorrhage. STUDY DESIGN: Prospective study. ANIMALS: Twelve dogs. METHODS: Dogs undergoing surgical oncology procedures associated with a high risk of hemorrhage were enrolled. Blood was collected a minimum of 6 days before surgery and separated into fresh frozen plasma (FFP) and packed red blood cells (pRBC). Dogs received FFP at the start of surgery and pRBC intraoperatively when hemorrhage ensued. The mean packed cell volume/total solids (PCV/TS) were calculated on the day of PABD preoperatively, immediately postoperatively, and 24 hours after transfusion. The dogs were monitored for transfusion-related adverse reactions, including hyperthermia, hypotension, tachycardia, bradycardia, pale mucous membranes, prolonged capillary refill time, or tachypnea/dyspnea. RESULTS: Dogs enrolled in the study underwent mandibulectomy, maxillectomy, chest wall resection, and liver lobectomy. Ten of the 12 dogs that underwent PABD received autologous transfusion at first signs of hemorrhage intraoperatively. Iatrogenic anemia was noted in two dogs (PCV 30% and 31%). The mean PCV/TS levels on the day of blood collection, preoperatively, immediately postoperatively (after transfusion), and 24 hours posttransfusion were 45.1%/7.1 g/dL, 42.2%/6.73 g/dL, 33.2%/5.42 g/dL, and 36.5%/5.65 g/dL, respectively. No dog developed transfusion-related complications. CONCLUSION: Preoperative autologous blood donation was well tolerated and led to uneventful autologous transfusion in 10 of 12 dogs. CLINICAL SIGNIFICANCE: Preoperative autologous blood donation and autologous transfusion are feasible for dogs undergoing elective surgical procedures with a high risk of hemorrhage.

Cutaneous Signs in Dermatological Diseases; An Overview.

Cutaneous signs in dermatology are pathognomonic or diagnostic for certain conditions. A knowledge of these signs is a useful skill, which must not be forgotten in the recent trend towards relying on molecular biology and other investigative tools including histological findings to make a diagnosis. The aim of this review is to summarize all cutaneous signs in dermatology in a systematic way that would benefit dermatologists, trainee or experienced, in identifying diseases and sharpening their clinical skills. A keyword search for the terms "cutaneous signs," "sign" AND "dermatology" was conducted through PUBMED, Google, and the major textbooks in dermatology (i.e., Rooks Textbook of Dermatology; Fitzpatrick's Dermatology in General Medicine and Dermatology). References were searched thoroughly for all cutaneous signs described. One hundred and nineteen signs were noted and summarized under the following categories: autoimmune, infective, inflammatory, neoplastic, genetic, trauma/miscellaneous. This review would serve as a good reference for those wanting to improve their clinical acumen in diagnosing dermatological disease.

Osteosarcoma of the proximal tibia in a dog 6 years after tibial tuberosity advancement.

A 9-year-old spayed female Cane Corso dog was presented with a 1-week history of an acute right hind limb non-weight-bearing lameness. Six years previously, a right tibial tuberosity advancement (TTA) procedure had been performed. Orthogonal radiographs of the right pelvic limb taken before presentation revealed a comminuted displaced mid-diaphyseal tibial fracture. An aggressive bone lesion with osteolysis and periosteal reaction of the proximal tibia were evident that were most consistent with a primary bone tumor. The owner elected to have the dog euthanized. After euthanasia a fine-needle aspirate of the right proximal tibia was submitted for clinical pathology. Cytology and alkaline phosphatase staining of the fine-needle aspirate were consistent with osteosarcoma. Key clinical message: To the authors' knowledge, this is only the second reported case of osteosarcoma at the site of a TTA. Implantassociated osteosarcoma has been associated with tibial plateau levelling osteotomy (TPLO) and fracture repair.

Une femelle Cane Corso stérilisée âgée de 9 ans fut présentée avec une histoire d'une boiterie sans appui de la patte arrière droite apparue de manière aiguë depuis 1 semaine. Six ans auparavant, une chirurgie d'avancement de la tubérosité tibiale droite (TTA) avait été réalisée. Des radiographies orthogonales du membre pelvien droit prises avant la présentation ont révélé une fracture comminutive déplacée du tibia au niveau mi-diaphysaire. Une lésion osseuse agressive avec ostéolyse et réaction du périoste du tibia proximal étaient évidentes et considérées compatibles avec une tumeur osseuse primaire. Le propriétaire opta pour l'euthanasie du chien. À la suite de l'euthanasie une aspiration à l'aiguille fine du tibia proximal droit fut soumise pour examen en pathologie clinique. La cytologie et une coloration à la phosphatase alcaline de l'aspiration à l'aiguille fine étaient compatibles avec un ostéosarcome.Message clinique clé :À la connaissance de l'auteur, ceci est seulement le deuxième cas rapporté d'un ostéosarcome au site d'une TTA. Les ostéosarcomes associés aux implants ont été associés avec l'ostéotome de nivellement du plateau tibial (TPLO) et la réparation de fracture.(Traduit par Dr Serge Messier).

Parenteral administration of factor Xa/IIa inhibitors limits experimental aortic aneurysm and atherosclerosis.

Intraluminal thrombus is a consistent feature of human abdominal aortic aneurysm (AAA). Coagulation factor Xa (FXa) catalyses FII to thrombin (FIIa). We examined the effect of FXa/FIIa inhibition on experimental aortic aneurysm in apolipoprotein E-deficient (ApoE-/-) mice infused with angiotensin II (AngII). The concentration of FXa within the supra-renal aorta (SRA) correlated positively with SRA diameter. Parenteral administration of enoxaparin (FXa/IIa inhibitor) and fondaparinux (FXa inhibitor) over 14 days reduced to severity of aortic aneurysm and atherosclerosis in AngII-infused ApoE-/- mice. Enteral administration of the FIIa inhibitor dabigatran had no significant effect. Aortic protease-activated receptor (PAR)-2 expression increased in response to AngII infusion. Fondaparinux reduced SRA levels of FXa, FIIa, PAR-2, matrix metalloproteinase (MMP)2, Smad2/3 phosphorylation, and MOMA-2 positive cells in the mouse model. FXa stimulated Smad2/3 phosphorylation and MMP2 expression in aortic vascular smooth muscle cells (VSMC) in vitro. Expression of MMP2 in FXa-stimulated VSMC was downregulated in the presence of a PAR-2 but not a PAR-1 inhibitor. These findings suggest that FXa/FIIa inhibition limits aortic aneurysm and atherosclerosis severity due to down-regulation of vascular PAR-2-mediated Smad2/3 signalling and MMP2 expression. Inhibition of FXa/FIIa may be a potential therapy for limiting aortic aneurysm.

Breast Cancer Awareness and Prevention Behavior Among Women of Delhi, India: Identifying Barriers to Early Detection.

BACKGROUND: Globally, breast cancer (BC) has become the leading cause of mortality in women. Awareness and early detection can curb the growing burden of BC and are the first step in the battle against BC. The aim of this qualitative study was to explore the awareness and perceived barriers concerning the early detection of BC. METHODS: A total of 20 focus group discussions (FGDs) were conducted during May 2013-March 2014. Pre-existing themes were used to conduct FGDs; each FGD group consisted of an average of ~10 women (aged ≥18-70 years) who came to participate in a BC awareness workshop. All FGDs were audio taped and transcribed verbatim. The transcripts were inductively analyzed using ATLAS.ti. Based on emerged codes and categories, thematic analysis was done, and theory was developed using the grounded theory approach. RESULTS: Data were analyzed in three major themes: i) knowledge and perception about BC; ii) barriers faced by women in the early presentation of BC; and iii) healthcare-seeking behavior. The findings revealed that shyness, fear, and posteriority were the major behavioral barriers in the early presentation of BC. Erroneously, pain was considered as an initial symptom of BC by most women. Financial constraint was also mentioned as a cause for delay in accessing treatment. Social stigma that breast problems reflect bad character of women also contributed in hiding BC symptoms. CONCLUSIONS: Lack of BC awareness was prevalent, especially in low socioeconomic class. Women's ambivalence in prioritizing their own health and social and behavioral hurdles should be addressed by BC awareness campaigns appropriately suited for various levels of social class.