Background: Thermal traumas impose a huge burden on healthcare systems. This merits the need for advanced but cost-effective remedies with clinical prospects. In this context, we prepared a regenerative 3D-construct comprising of Cassia angustifolia extract (SM) primed adipose-derived stem cells (ASCs) laden amniotic membrane for faster burn wound repair. Methods: ASCs were preconditioned with SM (30 µg/ml for 24 h), and subsequently exposed to in-vitro thermal injury (51 °C,10 min). In-vivo thermal injury was induced by placing pre-heated copper-disc (2 cm diameter) on dorsum of the Wistar rats. ASCs (2.0 × 105) pre-treated with SM (SM-ASCs), cultured on stromal side of amniotic membrane (AM) were transplanted in rat heat-injury model. Non-transplanted heat-injured rats and non-heat-injured rats were kept as controls. Results: The significantly upregulated expression of IGF1, SDF1A, TGFβ1, VEGF, GSS, GSR, IL4, BCL2 genes and downregulation of BAX, IL6, TNFα, and NFkB1 in SM-ASCs in in-vitro and in-vivo settings confirmed its potential in promoting cell-proliferation, migration, angiogenesis, antioxidant, cell-survival, anti-inflammatory, and wound healing activity. Moreover, SM-ASCs induced early wound closure, better architecture, normal epidermal thickness, orderly-arranged collagen fibers, and well-developed skin appendages in healed rat-skin transplanted with AM+SM-ASCs, additionally confirmed by increased expression of structural genes (Krt1, Krt8, Krt19, Desmin, Vimentin, α-Sma) in comparison to untreated-ASCs laden-AM transplanted in heat injured rats. Conclusion SM priming effectively enabled ASCs to counter thermal injury by significantly enhancing cell survival and reducing inflammation upon transplantation. This study provides bases for development of effective combinational therapies (natural scaffold, medicine, and stem cells) with clinical prospects for treating burn wounds.

Asthenoteratozoospermia is one of the most severe types of qualitative sperm defects. Most cases are due to mutations in genes encoding the components of sperm flagella, which have an ultrastructure similar to that of motile cilia. Coiled-coil domain containing 103 (CCDC103) is an outer dynein arm assembly factor, and pathogenic variants of CCDC103 cause primary ciliary dyskinesia (PCD). However, whether CCDC103 pathogenic variants cause severe asthenoteratozoospermia has yet to be determined. Whole-exome sequencing (WES) was performed for two individuals with nonsyndromic asthenoteratozoospermia in a consanguineous family. A homozygous CCDC103 variant segregating recessively with an infertility phenotype was identified (ENST00000035776.2, c.461A>C, p.His154Pro). CCDC103 p.His154Pro was previously reported as a high prevalence mutation causing PCD, though the reproductive phenotype of these PCD individuals is unknown. Transmission electron microscopy (TEM) of affected individuals' spermatozoa showed that the mid-piece was severely damaged with disorganized dynein arms, similar to the abnormal ultrastructure of respiratory ciliary of PCD individuals with the same mutation. Thus, our findings expand the phenotype spectrum of CCDC103 p.His154Pro as a novel pathogenic gene for nonsyndromic asthenospermia.
Asthenozoospermia/pathology* ; Dyneins/genetics* ; Homozygote ; Humans ; Male ; Microtubule-Associated Proteins ; Mutation ; Mutation, Missense ; Sperm Tail/metabolism*

COVID-19 ; Delivery of Health Care ; Humans ; Pakistan ; SARS-CoV-2 ; Telemedicine

BACKGROUND: Periploca aphylla is used by local population and indigenous medicine practitioners as stomachic, tonic, antitumor, antiulcer, and for treatment of inflammatory disorders. The aim of this study was to evaluate antidiabetic effect of the extract of P. aphylla and to investigate antioxidant and hypolipidemic activity in streptozotocin (STZ)-induced diabetic rats. METHODS: The present research was conducted to evaluate the antihyperglycemic potential of methanol extract of P. aphylla (PAM) and subfractions n-hexane (PAH), chloroform (PAC), ethyl acetate (PAE), n-butanol (PAB), and aqueous (PAA) in glucose-overloaded hyperglycemic Sprague-Dawley rats. Based on the efficacy, PAB (200 mg/kg and 400 mg/kg) was tested for its antidiabetic activity in STZ-induced diabetic rats. Diabetes was induced via intraperitoneal injection of STZ (55 mg/kg) in rat. Blood glucose values were taken weekly. HPLC-DAD analysis of PAB was carried out for the presence of various polyphenols. RESULTS: HPLC-DAD analysis of PAB recorded the presence of rutin, catechin, caffeic acid, and myricetin. Oral administration of PAB at doses of 200 and 400 mg/kg for 21 days significantly restored (P < 0.01) body weight (%) and relative liver and relative kidney weight of diabetic rats. Diabetic control rats showed significant elevation (P < 0.01) of AST, ALT, ALP, LDH, total cholesterol, triglycerides, LDL, creatinine, total bilirubin, and BUN while reduced (P < 0.01) level of glucose, total protein, albumin, insulin, and HDL in serum. Count of blood cells and hematological parameters were altered in diabetic rats. Further, glutathione peroxidase, catalase, superoxide dismutase, glutathione reductase, and total soluble protein concentration decreased while concentration of thiobarbituric acid reactive substances and percent DNA damages increased (P < 0.01) in liver and renal tissues of diabetic rats. Histopathological damage scores increased in liver and kidney tissues of diabetic rats. Intake of PAB (400 mg/kg) resulted in significant improvement (P < 0.01) of above parameters, and results were comparable to that of standard drug glibenclamide. CONCLUSION The result suggests the antihyperglycemic, antioxidant, and anti-inflammatory activities of PAB treatment in STZ-compelled diabetic rat. PAB might be used as new therapeutic agent in diabetic patients to manage diabetes and decrease the complications.
1-Butanol/chemistry* ; Administration, Oral ; Animals ; Diabetes Mellitus, Experimental/drug therapy* ; Dose-Response Relationship, Drug ; Hypoglycemic Agents/chemistry* ; Male ; Periploca/chemistry* ; Phytochemicals/chemistry* ; Plant Extracts/chemistry* ; Rats ; Rats, Sprague-Dawley ; Streptozocin/adverse effects*

BACKGROUND: Pilea umbrosa (Urticaceae) is used by local communities (district Abbotabad) for liver disorders, as anticancer, in rheumatism and in skin disorders. METHODS: Methanol extract of P. umbrosa (PUM) was investigated for the presence of polyphenolic constituents by HPLC-DAD analysis. PUM (150 mg/kg and 300 mg/kg) was administered on alternate days for eight weeks in rats exposed with carbon tetrachloride (CCl). Serum analysis was performed for liver function tests while in liver tissues level of antioxidant enzymes and biochemical markers were also studied. In addition, semi quantitative estimation of antioxidant genes, endoplasmic reticulum (ER) induced stress markers, pro-inflammatory cytokines and fibrosis related genes were carried out on liver tissues by RT-PCR analysis. Liver tissues were also studied for histopathological injuries. RESULTS: Level of antioxidant enzymes such as catalase (CAT), superoxide dismutase (SOD), peroxidase (POD) and glutathione (GSH) decreased (p < 0.05) whereas level of thiobarbituric acid reactive substance (TBARS), HO and nitrite increased in liver tissues of CCl treated rat. Likewise increase in the level of serum markers; alanine transaminase (ALT), aspartate transaminase (AST), alkaline phosphatase (ALP) and total bilirubin was observed. Moreover, CCl caused many fold increase in expression of ER stress markers; glucose regulated protein (GRP-78), x-box binding protein1-total (XBP-1 t), x-box binding protein1-unspliced (XBP-1 u) and x-box binding protein1-spliced (XBP-1 s). The level of inflammatory mediators such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1) was aggregated whereas suppressed the level of antioxidant enzymes; γ-glutamylcysteine ligase (GCLC), protein disulfide isomerase (PDI) and nuclear erythroid 2 p45-related factor 2 (Nrf-2). Additionally, level of fibrosis markers; transforming growth factor-β (TGF-β), Smad-3 and collagen type 1 (Col1-α) increased with CCl induced liver toxicity. Histopathological scrutiny depicted damaged liver cells, neutrophils infiltration and dilated sinusoids in CCl intoxicated rats. PUM was enriched with rutin, catechin, caffeic acid and apigenin as evidenced by HPLC analysis. Simultaneous administration of PUM and CCl in rats retrieved the normal expression of these markers and prevented hepatic injuries. CONCLUSION Collectively these results suggest that PUM constituted of strong antioxidant chemicals and could be a potential therapeutic agent for stress related liver disorders.
Animals ; Carbon Tetrachloride ; adverse effects ; Chemical and Drug Induced Liver Injury ; drug therapy ; etiology ; pathology ; Endoplasmic Reticulum Stress ; drug effects ; Fibrosis ; drug therapy ; genetics ; Inflammation ; drug therapy ; genetics ; Liver ; drug effects ; enzymology ; metabolism ; Male ; Protective Agents ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Urticaceae ; chemistry

Rationale: Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) has been recognized as highly pathogenic. The current pandemic of SARS-CoV-2 has been spread globally and infected more than 200 countries. Patient concerns: We report the first confirmed fatal case of COVID-19 in Pakistan. A 50-year-old man returned from Saudi Arabia on March 09, 2020 and presented with cough, fever, malaise, poor appetite and difficulty in breathing to the Pulmonologist at District Headquarter Hospital Mardan. Diagnosis: The patient was initially diagnosed as COVID-19 suspected case. A oropharyngeal swab sample was positive by realtime RT-PCR tests. Lessons: This report highlights the importance of close coordination between clinicians and public health authorities as well as the importance of early laboratory-based confirmation of COVID-19 cases.

Objective: To explore potential inhibitors of viral enzymes of SARS CoV-2. Methods: The in-silico docked potential of anti-viral, antibiotic, and analgesic drugs were studied for inhibition of the nonstructural protein (NSP) 9, NSP3, and NSP15 of SARS CoV-2 using recent structural peculiarities of these enzymes, 3D optimized structures of drugs and algorithm-based ligand inhibitory potential. Results: Teicoplanin, azithromycin, and remdesivir potentially inhibited NSP9 (Dock-score 9 620, 5 472 and 6 252, respectively), NSP3 (Dock-score 9 846, 5 604 and 5 548, respectively) and NSP15 (Dock-score 10 960, 6414 and 6 002, respectively). Conclusions: Teicoplanin acts as a significant receptor antagonist and potentially inhibits the SARS CoV-2 enzymes.

No abstract available.
Humans ; Ichthyosis ; Ichthyosis, Lamellar

Impalement injuries, is a severe form of trauma, which are not common in civilian life. These injuries rarely occurs in major accidents. Abdomen, chest, limbs and perineum are often involved due to their large surface area. Thoracic impalement injury is usually a fatal injury, due to location of major vessels and heart in the thoracic cavity. These injuries are horrifying to site, but the patients who are lucky enough to make it to hospital, usually survive. Chances of survival are larger in right sided impalement injuries while central injuries are always died at the scene. Our patient, 25 years old male, was brought to the emergency room (ER) with large impaled metallic bar (about 2.5 feet long) in situ, in right sided chest. The patient was immediately shifted to operation room (OR) and was operated, his recovery was uneventful without any sequelae. Such patients should be treated and resuscitated according to advanced trauma life support (ATLS) protocols and operated without any delay for further investigations. Such operations are carried out by the most experienced surgeon team available. The impaled objects should not be processed if not necessary to avoid major hemorrhage and damage to vital structures, until the patient is in operation room. Large size and unusual position of impaled objects, makes the job difficult for surgeons/anesthetists. Although horrifying at scene, patients with thoracic impalement injuries are mostly young and healthy, and those who survive the pre-hospital phase are potentially manageable with proper resuscitation. Usually these patients make recovery without any further complications.
Accidents, Traffic ; Adult ; Advanced Trauma Life Support Care ; Foreign Bodies ; surgery ; Humans ; Male ; Metals ; Resuscitation ; Survivors ; Thoracic Injuries ; surgery ; Thoracic Surgical Procedures ; methods ; Thorax ; Treatment Outcome ; Wounds, Penetrating ; surgery