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PURPOSE: Pediatric cholestasis is the phenotypic expression of clinically and genetically heterogeneous disorders of bile acid synthesis and flow. Although a growing number of monogenic causes of pediatric cholestasis have been identified, the majority of cases remain undiagnosed molecularly. METHODS: In a cohort of 299 pediatric participants (279 families) with intrahepatic cholestasis, we performed exome sequencing as a first-tier diagnostic test. RESULTS: A likely causal variant was identified in 135 families (48.56%). These comprise 135 families that harbor variants spanning 37 genes with established or tentative links to cholestasis. In addition, we propose a novel candidate gene (PSKH1) (HGNC:9529) in 4 families. PSKH1 was particularly compelling because of strong linkage in three consanguineous families who shared a novel hepatorenal ciliopathy phenotype. Two of the four families shared a founder homozygous variant while the third had a different homozygous variant in PSKH1. PSKH1 encodes a putative protein serine kinase of unknown function. Patient fibroblasts displayed abnormal cilia that are long and show abnormal transport. A homozygous Pskh1 mutant mouse faithfully recapitulated the human phenotype and displayed abnormally long cilia. The phenotype could be rationalized by the loss of catalytic activity observed for each recombinant PSKH1 variant using in vitro kinase assays. CONCLUSION: Our results support the use of genomics in the workup of pediatric cholestasis and reveal PSKH1-related hepatorenal ciliopathy as a novel candidate monogenic form.
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OBJECTIVE: To assess the outcomes of low-risk differentiated thyroid cancer (DTC) with tumour size 1 to 4 cm post-thyroidectomy in patients who never received radioactive iodine (RAI) ablation and to compare the outcomes with those who received RAI therapy. STUDY DESIGN: Observational study. Place and Duration of the Study: Department of Nuclear Medicine, Karachi Institute of Radiotherapy and Nuclear Medicine (KIRAN) Hospital, Karachi, Pakistan, from January 2016 to December 2020. METHODOLOGY: A total of 130 consecutive patients of low-risk DTC of both genders aged between 18-75 years were identified who underwent thyroidectomy. Patients were classified as either being treated or not treated with RAI, based on RAI data post-thyroidectomy. Patients were followed up for two to five years depending on their date of diagnosis from 2016 to 2020, using thyroglobulin (Tg), anti-thyroglobulin (anti-Tg), and thyrotropin (TSH) levels and ultrasound neck. Association between patients who received RAI and who did not receive RAI was determined and a p-value <0.05 was considered as statistical significance. RESULTS: Patients had mean age of 34.5 ± 10.4 years with majority of females (75.4%). Majority of the patients (94.6%) had total thyroidectomy with no neck dissection (83.1%). RAI therapy was conducted among 70.8% participants, of which, 56.9% participants received a dose of 100 mci. Most of the patients had positive outcomes as there was no recurrence among 96.2% participants. There was a significant difference in RAI therapy with total thyroidectomy as compared to subtotal, RAI dose, tumour stage, neck dissection, and lymph node involvement (p ≤0.001). CONCLUSION: After complete tumour resection, a highly selected group of patients with low-risk local disease have shown low rate of recurrence when managed without RAI. Interestingly, the disease recurrence was also only seen in patients who received RAI therapy in comparison to those who did not receive RAI therapy. KEY WORDS: Outcomes, Differentiated thyroid cancer, Radioactive iodine, Ablation therapy, Post-surgery.
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Radioisótopos do Iodo , Neoplasias da Glândula Tireoide , Tireoidectomia , Humanos , Feminino , Neoplasias da Glândula Tireoide/cirurgia , Neoplasias da Glândula Tireoide/radioterapia , Neoplasias da Glândula Tireoide/patologia , Masculino , Adulto , Pessoa de Meia-Idade , Paquistão/epidemiologia , Radioisótopos do Iodo/uso terapêutico , Resultado do Tratamento , Adolescente , Idoso , Adulto Jovem , Recidiva Local de Neoplasia , Centros de Atenção TerciáriaRESUMO
Huntington's disease (HD) is an autosomal dominant disease caused by the expansion of cytosine-adenine-guanine (CAG) repeats in one copy of the HTT gene (mutant HTT, mHTT). The unaffected HTT gene encodes wild-type HTT (wtHTT) protein, which supports processes important for the health and function of the central nervous system. Selective lowering of mHTT for the treatment of HD may provide a benefit over nonselective HTT-lowering approaches, as it aims to preserve the beneficial activities of wtHTT. Targeting a heterozygous single-nucleotide polymorphism (SNP) where the targeted variant is on the mHTT gene is one strategy for achieving allele-selective activity. Herein, we investigated whether stereopure phosphorothioate (PS)- and phosphoryl guanidine (PN)-containing oligonucleotides can direct allele-selective mHTT lowering by targeting rs362273 (SNP3). We demonstrate that our SNP3-targeting molecules are potent, durable, and selective for mHTT in vitro and in vivo in mouse models. Through comparisons with a surrogate for the nonselective investigational compound tominersen, we also demonstrate that allele-selective molecules display equivalent potency toward mHTT with improved durability while sparing wtHTT. Our preclinical findings support the advancement of WVE-003, an investigational allele-selective compound currently in clinical testing (NCT05032196) for the treatment of patients with HD.
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Introduction: Achromobacter xylosoxidans is an uncommon opportunistic organism, mainly causing infection in immune-compromised hosts. A. xylosoxidans is a non-fermenting Gram-negative bacillus. Being closely associated with Alcaligenes species was also called A. xylosoxidans. Case Report: A few cases of periprosthetic infection by A. xylosoxidans have been reported outside India. A periprosthetic infection with A. xylosoxidans following a total knee arthroplasty is reported in a female. Conclusion: A. xylosoxidans is a pathogen capable of causing prosthetic joint infection even in immunocompetent patients. Thorough debridement and appropriate antibiotic treatment is essential for the success of revision surgery.
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BACKGROUND AND PURPOSE: This prospective observational study aimed to investigate the occurrence of avascular necrosis (AVN) of the femoral head in COVID-19 patients through MRI scans. The study examined the patterns of AVN in 110 individuals who had undergone conventional COVID-19 therapy and reported hip discomfort. This study highlights the importance of considering AVN as a potential complication of COVID-19 therapy, particularly in younger patients who experience hip discomfort. METHODS: Individuals who had corticosteroid treatment for COVID-19 and experienced hip discomfort during 6 months between January 2022 and August 2022 were included in this study, and an MRI scan was done to observe changes in the hip joint. RESULTS: The results were classified using the Ficat and Arlet classification system. The analysis revealed that AVN was not present in 91.81% of cases. However, Stage I AVN was detected in 4.54% of cases, Stage II AVN in 2.72% of cases, and Stage III AVN in 1.1% of cases. No cases of Stage IV AVN were observed. CONCLUSIONS: The study concludes that AVN occurred in 6% of individuals who underwent conventional therapy for COVID-19 and experienced hip discomfort. In these settings (post COVID-19), normal MRI results were more typical, and mild AVN (Stage I) was a frequent finding in MRI scans that were positive.
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COVID-19 , Necrose da Cabeça do Fêmur , Imageamento por Ressonância Magnética , Humanos , Necrose da Cabeça do Fêmur/diagnóstico por imagem , Necrose da Cabeça do Fêmur/etiologia , Masculino , Feminino , COVID-19/complicações , COVID-19/diagnóstico por imagem , Imageamento por Ressonância Magnética/métodos , Pessoa de Meia-Idade , Estudos Prospectivos , Adulto , Idoso , Tratamento Farmacológico da COVID-19RESUMO
The stacking 2D materials, such as molybdenum disulfide (MoS2), are among the most promising candidates for detecting H2S gas. Herein, we designed a series of novel nanocomposites consisting of MoS2 and ZnCo2O4. These materials were synthesized via a simple hydrothermal method. The microstructure and morphology of nanocomposites were studied by different characteristics such as X-ray diffraction, scanning electron microscopy, energy-dispersive X-ray spectroscopy, transmission electron microscopy, high-resolution transmission electron microscopy, Brunauer-Emmett-Teller (BET), and X-ray photoelectron spectroscopy. These nanocomposites were used as gas sensors, and the highest response (6.6) toward 10 ppm of H2S was detected by the gas sensor of MZCO-6 (having MoS2 contents 0.060 g) among all other tested sensors. The response value (Ra/Rg) was almost three times that of pure ZnCo2O4 (Ra/Rg = 2). In addition, the sensor of MZCO-6 exposed good selectivity, short response/recovery time (12/28 s), long-term stability (28 days), and a low detection limit (0.5 ppm) toward H2S gas at RT. The excellent performance of MZCO-6 may be attributed to some features of MoS2, such as stack structure, higher BET and surface area and active sites, a synergistic effect, etc. This simple fabrication sensor provides a novel idea for detecting H2S gas at RT.
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In a continuous search for an essential antidiabetic agent, Sida cordifolia hydroalcoholic (SCHA) extract-loaded chitosan nanoparticles (SCHA-CS-NP) were optimized. The Box-Behnken design (BBD Design-Expert software, version 14) with three parameters was used to optimize the nanoparticles after creating them using the ion gelation method. The chitosan and Tween 20 contents and the stirring speed were chosen as the independent variables, and their separate and combined effects on particle size (Y1), polydispersity index (Y2) and entrapment efficiency (Y3) were observed. The optimized formulation showed a particle size of 51 nm, an entrapment efficiency of 84.54% and a polydispersity index of 0.391. Physicochemical characterization, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), a drug release study, an ex vivo permeation study, and an antioxidant study were performed. Confocal laser scanning microscopy (CLSM) images demonstrated that chitosan nanoparticles loaded with rhodamine B-laden SCHA extract had superior penetration compared to the control (rhodamine B solution). Furthermore, compared to conventional ascorbic acid (IC50 = 45 µg/mL), a superior antioxidant activity was discovered for SCHA-CS-NPs (IC50 = 86.45 ± 2.24 µg/mL), while SCHA-CS-NPs also exhibited strong antidiabetic potential (IC50 = 93.71 ± 1.79 µg/mL) compared to standard acarbose (IC50 = 97.25 ± 1.43 µg/mL). The overall results demonstrated that SCHA-CS-NPs are a promising and efficient formulation for oral delivery.
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Persistent Müllerian Duct syndrome is a rare male disorder of sexual development. The phenotypically and genotypically male patient presents with female internal organs (i.e., uterus, cervix, fallopian tubes and upper part of vagina) due to deficiency of anti-mullerian hormone or insensitivity of tissues to Anti Mullerian Hormone. We present a 19 year old male who came with complaint of right iliac fossa pain. He was investigated for acute appendicitis and on imaging, he was diagnosed to have bilateral cryptorchidism with rudimentary uterus. Computed tomography followed by pelvic ultrasonography was done which indicated two testes in abdomen and a soft tissue density structure, identified as a rudimentary uterus located posterior to the urinary bladder. CT scan findings were further confirmed by magnetic resonance imaging pelvis. A trial of stepwise orchidopexy followed by orchidectomy with removal of rudimentary uterus was performed laparoscopically. Additionally, he was counselled for long term sex hormone replacement and reproductive failure in future.
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Criptorquidismo , Transtorno 46,XY do Desenvolvimento Sexual , Humanos , Masculino , Feminino , Adulto Jovem , Adulto , Transtorno 46,XY do Desenvolvimento Sexual/diagnóstico , Transtorno 46,XY do Desenvolvimento Sexual/cirurgia , Criptorquidismo/diagnóstico , Criptorquidismo/cirurgia , Hormônio Antimülleriano , Ductos Paramesonéfricos/cirurgia , Ductos Paramesonéfricos/patologiaRESUMO
Solanum xanthocarpum (SX) has been used to treat a variety of diseases, including skin disorders like psoriasis (PSO). SX possesses many pharmacological activities of anti-inflammatory, anti-cancer, immunosuppressive, and healing qualities. However, the multi-target mechanism of SX on PSO still needs clarity. Materials and methods: The Indian Medicinal Plants, Phytochemicals and Therapeutics (IMPPAT) database and the Swiss Target Prediction online tool were used to find the active phytochemical components and their associated target proteins. OMIM and GeneCards databases were used to extract PSO-related targets. A Venn diagram analysis determined the common targets of SX against PSO. Subsequently, the protein-protein interaction (PPI) network and core PPI target analysis were carried out using the STRING network and Cytoscape software. Also, utilising the online Metascape and bioinformatics platform tool, a pathway enrichment analysis of common targets using the Kyoto Encyclopaedia of Genes and Genome (KEGG) and Gene Ontology (GO) databases was conducted to verify the role of targets in biological processes, cellular components and molecular functions with respect to KEGG pathways. Lastly, molecular docking simulations were performed to validate the strong affinity between components of SX and key target receptors. Results: According to the IMPPAT Database information, 8 active SX against PSO components were active. According to the PPI network and core targets study, the main targets against PSO were EGFR, SRC, STAT3, ERBB2, PTK2, SYK, EP300, CBL, TP53, and AR. Moreover, molecular docking simulations verified the binding interaction of phytochemical SX components with their PSO targets. Last but not least, enrichment analysis showed that SX is involved in several biological processes, including peptidyl-tyrosine phosphorylation, peptidyl-tyrosine modification, and peptidyl-serine modification. The relevant KEGG signalling pathways are the PI3K-AKT signalling pathway, the EGFR tyrosine kinase inhibitor resistance pathway, and the MAPK signalling pathway. Conclusion: The network pharmacology technique, which is based on data interpretation and molecular docking simulation techniques, has proven the multi-target function of SX phytoconstituents.
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In this study, hesperidin was loaded into a transethosome and was developed employing the rotary evaporator method. The formulation was optimized using the Box-Behnken design (BBD). The optimized HSD-TE formulation has a spherical shape, vesicle size, polydispersity index, entrapment efficiency, and zeta potential within the range of 178.98 nm; the PDI was 0.259 with a zeta potential of -31.14 mV and % EE of 89.51%, respectively. The in vitro drug release shows that HSD-TE exhibited the release of 81.124 ± 3.45% in comparison to HSD suspension. The ex vivo skin permeation showed a 2-fold increase in HSD-TE gel permeation. The antioxidant activity of HSD-TE was found to be 79.20 ± 1.77% higher than that of the HSD solution. The formulation showed 2-fold deeper HSD-TE penetration across excised rat skin membranes in confocal laser microscopy scanning, indicating promising in vivo prospects. In a dermatokinetic study, HSD-TE gel was compared to HSD conventional gel where TE significantly boosted HSD transport in the epidermis and dermal layers. The formulation showed greater efficacy than free HSD in the inhibition of microbial growth, as evidenced by antibacterial activity on the Gram-negative and positive bacteria. These investigations found that the HSD-TE formulation could enhance the topical application in the management of cutaneous bacterial infections.
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Strychnine (STCN) has demonstrated an exceptional anticancer effect against various cancers. However, the STCN clinical utility has been hampered by its low water solubility, restricted therapeutic window, short half-life, and significant toxicity. The objective of this investigation was to design and optimize a formulation of strychnine-loaded transliposomes (STCN-TLs) for dermal administration of STCN to treat skin cancer. The formulations of STCN-TL were examined in terms of vesicle size (VS), polydispersity index (PDI), entrapment efficiency (EE), and in vitro delivery. The improved STCN-TL formulation exhibited VS, PDI, EE, and in vitro delivery of 101.5 ± 2.14 nm, 0.218 ± 0.12, 81.74 ± 1.43%, and 85.39 ± 2.33%, respectively. In an ex vivo penetration, the created STCN-TL formulation demonstrated a 2.5-fold increase in permeability compared to the STCN solution. CLSM pictures of skin (rat) revealed that the rhodamine B-loaded transliposome preparation penetrated deeper than the rhodamine B hydroalcoholic mixture. Additionally, rat skin managed with STCN-TL nanogel exhibited a significant increase in Cskin max and AUC0-8 compared to rat skin treated with traditional STCN gel. The findings demonstrated that the transliposome preparation might be a suitable nanocarrier for the cutaneous distribution of STCN in the amelioration of skin cancer.
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The layered 2-D materials, such as molybdenum disulfide (MoS2), are among the most promising candidates for detecting H2S gas at very low concentrations. Herein, we have designed a series of novel nanocomposites consisting of MoS2 and NiO. These materials were synthesized via a simple hydrothermal method. The microstructure and morphology of nanocomposites were studied using different characterization techniques, such as X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), high-resolution transmission electron microscopy (HRTEM), Brunauer-Emmett-Teller (BET) analysis, and X-ray photoelectron spectroscopy (XPS). These nanocomposites were used as gas sensors, and the highest response (6.3) towards 10 ppm H2S was detected by the MNO-10 gas sensor among all the tested sensors. The response value (Rg/Ra) was almost three times that of pure NiO (Rg/Ra = 2). Besides, the MNO-10 sensor exposed good selectivity, short response/recovery time (50/20 s), long-term stability (28 days), reproducibility (6 cycles), and a low detection limit (2 ppm) towards H2S gas at RT. The excellent performance of MNO-10 may be attributed to some features of MoS2, such as a layered structure, higher BET surface area, higher active sites, and a synergistic effect between MoS2 and NiO. This simple fabrication sensor throws a novel idea for detecting H2S gas.
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Hydrogen sulfide (H2S) detection is extremely necessary due to its hazardous nature. Thus, the design of novel sensors to detect H2S gas at low temperatures is highly desirable. In this study, a series of nanocomposites based on MoS2 octahedrons and ZnO-Zn2SnO4 nanoparticles were synthesized through the hydrothermal method. Various characterizations such as X-ray diffraction (XRD), Brunauer-Emmett-Teller (BET), scanning electron microscopy (SEM), transmission electron microscopy (TEM), energy-dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectrum (XPS) have been used to verify the crystal phase, morphology and composition of synthesized nanocomposites. Three gas sensors based on the nanocomposites of pure ZnO-Zn2SnO4 (MS-ZNO-0), 5 wt% MoS2-ZnO-Zn2SnO4 (MS-ZNO-5) and 10 wt% MoS2-ZnO-Zn2SnO4 (MS-ZNO-10) were fabricated to check the gas sensing properties of various volatile organic compounds (VOCs). It showed that the gas sensor of (MS-ZNO-5) displayed the highest response of 4 to 2 ppm H2S and fewer responses to all other tested gases at 30 °C. The sensor of MS-ZNO-5 also displayed humble selectivity (1.6), good stability (35 days), promising reproducibility (5 cycles), rapid response/recovery times (10 s/6 s), a limit of detection (LOD) of 0.05 ppm H2S (Ra/Rg = 1.8) and an almost linear relationship between H2S concentration and response. Several elements such as the structure of MoS2, higher BET-specific surface area, n-n junction and improvement in oxygen species corresponded to improving response.
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Lemborexant (LEM) is a novel dual orexin receptor antagonist (DORA), recently approved for the treatment of insomnia. As with other DORAs, LEM has potential of abuse and therefore placed in Schedule IV class by the United States Drug Enforcement Administration (USDEA). In this study, a sensitive and accurate UPLC-MS/MS assay was developed for the quantification of LEM in human plasma sample using losartan as an internal standard (IS). The chromatographic separation was performed by using gradient elution of mobile phase, comprising of 10 mM ammonium acetate and acetonitrile with a flow rate of 0.3 mL/min. An Acquity UPLC BEH C18 (1.7 µm, 2.1 × 50 mm) column was used for separation of LEM and IS by maintaining the oven temperature of 40 °C. The electrospray ionization in positive mode was used for sample ionization. The precursor to product ion transition of 411.12 > 175.09 (qualifier) and 411.1 > 287.14 (quantifier) was used for detection and quantification of LEM, respectively, in multiple reaction monitoring mode. Being a drug of abuse, the assay was validated according to "Scientific Working Group for Toxicology" (SWGTOX) guidelines, including limit of detection (LOD), limit of quantification (LOQ), precision and bias, calibration model, interferences, carry-over effects, matrix effects, and stability parameters. The LOD and LOQ of the assay were 0.35 and 1.0 ng/mL, respectively. The linear range was between 1-300 ng/mL with correlation coefficient of ≥0.995. The method was also cross validated in rat plasma samples with acceptable ranges of precision and accuracy before its application for pharmacokinetic study in rats.
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BACKGROUND: For the treatment of hypothyroidism, patients are usually placed on lifelong levothyroxine replacement therapy. Achieving clinical and biochemical euthyroid status is sometimes difficult because of several factors, including poor compliance or poor absorption of levothyroxine in the gut mucosa. CASE PRESENTATION: We present the case of a 45-year-old South Asian female with hypothyroidism on lifelong levothyroxine replacement. However, on her multiple follow-up visits over the past 2 years, it was noted that her thyrotropin level was never controlled and was not in range. A thorough history was unyielding in terms of compliance regarding levothyroxine medication and use of over-the-counter pills or mineral supplements that may interfere with the absorption of levothyroxine. Hence, we performed levothyroxine absorption test to rule out any malabsorption as well as document pseudo-malabsorption favoring drug nonadherence. Levothyroxine absorption test resulted in more than 56% absorption of levothyroxine; therefore, the patient was put on weekly directly observed treatment strategy resulting in normalization of thyrotropin levels on successive follow-ups. CONCLUSION: Directly observed treatment to replace levothyroxine once a week can be used as an alternative by health care professionals in patients in whom compliance to daily levothyroxine is the hidden cause of persistently raised thyrotropin. This strategy can reduce undue health burden on the patient and improve adherence to self-medication under confident supervision of the treating physician.
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Hipotireoidismo , Tireotropina , Humanos , Feminino , Pessoa de Meia-Idade , Tiroxina/uso terapêutico , Pacientes Ambulatoriais , Hipotireoidismo/tratamento farmacológico , Cooperação do PacienteRESUMO
Real-time sensing of hydrogen sulfide (H2S) at room temperature is important to ensure the safety of humans and the environment. Four kinds of different nanocomposites, such as MXene Ti3C2Tx, Ti3AlC2, WS2, and MoSe2/NiCo2O4, were synthesized using the hydrothermal method in this paper. Initially, the intrinsic properties of the synthesized nanocomposites were studied using different techniques. P-type butane and H2S-sensing behaviors of nanocomposites were performed and analyzed deeply. Four sensor sheets were fabricated using a spin-coating method. The gas sensor was distinctly part of the chemiresistor class. The MXene Ti3C2Tx/NiCo2O4-based gas sensor detected the highest response (16) toward 10 ppm H2S at room temperature. In comparison, the sensor detected the highest response (9.8) toward 4000 ppm butane at 90 °C compared with the other three fabricated sensors (Ti3AlC2, WS2, and MoSe2/NiCo2O4). The MXene Ti3C2Tx/NiCo2O4 sensor showed excellent responses, minimum limits of detection (0.1 ppm H2S and 5 ppm butane), long-term stability, and good reproducibility compared with the other fabricated sensors. The highest sensing properties toward H2S and butane were accredited to p-p heterojunctions, higher BET surface areas, increased oxygen species, etc. These simply synthesized nanocomposites and fabricated sensors present a novel method for tracing H2S and butane at the lowest concentration to prevent different gas-exposure-related diseases.
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INTRODUCTION: The primary aim of this study was to determine a plausible association between the radiological location of multiple sclerosis (MS) lesions and serum 25-hydroxyvitamin D (vitamin D) levels at the time of diagnosis. MS is a common immune-mediated neurological condition mainly affecting the central nervous system. Although the association of vitamin D levels is well established, there have not been many studies to propose a connection between the location of the lesions based on serum vitamin D levels. In this study, we determine the association between serum 25-hydroxyvitamin D and the radiological distribution of lesions in patients with MS. METHODS: Twenty patients with a confirmed diagnosis of MS involving new T2-weighted and gadolinium-enhancing T1-weighted lesions in the entire central nervous system (brain and spinal cord) with serum 25-hydroxyvitamin D levels at the time of diagnosis were included in a case group. As a reference, 20 patients with a confirmed diagnosis of MS with isolated new T2-weighted and gadolinium-enhancing T1-weighted lesions (either supratentorial, infratentorial, or spinal cord) with serum 25-hydroxyvitamin D levels at the time of diagnosis were included in the control group. RESULTS: The mean serum 25-hydroxyvitamin D level was significantly low in the case group compared to the control group (36.2 ± 17.2 vs 62.6 ± 21.0; p-value <0.0001). CONCLUSION: There is a plausible inverse relationship between serum vitamin D and the MS lesions involving the entire central nervous system (brain and spinal cord). This evidence may enable clinicians to forecast disease load based on serum vitamin D levels.
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Masson's Trichrome Staining (MTS) is a useful tool for analyzing fibrosis in a plethora of disease pathologies by differential staining of tissue components. It is used to identify collagen fibers in different tissues like heart, lung, skin, and muscles. Especially in cardiac fibrosis, MTS stains the collagen fibers (blue color), which helps in the distinction of scar area versus the healthy area (red color). However, there are several challenges to stain both paraffin-embedded sections and frozen (cryosections) using a single protocol. Therefore, the goal of this study was to develop a simple short protocol to assess cardiac fibrosis in both paraffin-embedded and cryo heart sections. MTS uses three different stains, i.e., Weigert's Iron Hematoxylin, Biebrich scarlet-acid fuchsin, and aniline blue to detect nuclei, cytoplasm, and collagen, respectively. In this study, we developed a simple short protocol that can be adapted by any lab to easily assess cardiac fibrosis in paraffin and frozen heart sections. Furthermore, we have addressed the challenges that are commonly faced during the immunostaining process and troubleshooting techniques. Overall, we have successfully developed a simple one-step protocol to assess myocardial fibrosis in paraffin-embedded and frozen cryosections.
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In this study, ultrasound-assisted extraction conditions were optimized to maximize the yields of sennoside A, sennoside B, aloe-emodin, emodin, and chrysophanol from S. alexandrina (aerial parts). The three UAE factors, extraction temperature (S1), extraction time (S2), and liquid to solid ratio (S3), were optimized using response surface methodology (RSM). A Box-Behnken design was used for experimental design and phytoconstituent analysis was performed using high-performance liquid chromatography-UV. The optimal extraction conditions were found to be a 64.2 °C extraction temperature, 52.1 min extraction time, and 25.2 mL/g liquid to solid ratio. The experimental values of sennoside A, sennoside B, aloe-emodin, emodin, and chrysophanol (2.237, 12.792, 2.457, 0.261, and 1.529%, respectively) agreed with those predicted (2.152, 12.031, 2.331, 0.214, and 1.411%, respectively) by RSM models, thus demonstrating the appropriateness of the model used and the accomplishment of RSM in optimizing the extraction conditions. Excellent antioxidant properties were exhibited by S. alexandrina methanol extract obtained using the optimized extraction conditions with a DPPH assay (IC50 = 59.7 ± 1.93, µg/mL) and ABTS method (47.2 ± 1.40, µg/mL) compared to standard ascorbic acid.
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Antioxidantes/farmacologia , Fracionamento Químico/métodos , Componentes Aéreos da Planta/química , Extrato de Senna/farmacologia , Senna/química , Sonicação , Ondas Ultrassônicas , Algoritmos , Antioxidantes/química , Antioxidantes/isolamento & purificação , Relação Dose-Resposta a Droga , Modelos Teóricos , Estrutura Molecular , Compostos Fitoquímicos , Extrato de Senna/química , Extrato de Senna/isolamento & purificaçãoRESUMO
BACKGROUND: Around 130 million infections of hepatitis C virus with 3% overall prevalence are there worldwide. There are approximately 4-5 million persons coinfected with HIV. The main objectives of this study were to determine the prevalence of HCV among HIV-positive individuals and to assess the predictors involved in the outcomes of HIV-HCV coinfected patients. METHODS: A retrospective, cross-sectional study was conducted on patients enrolled from 2007 to 2012 at Infectious Disease Unit, Hospital Palau Pinang, Pinang, Malaysia. Sociodemographic da%)ta as well as clinical data were collected with the help of a valid data collection form from the patients' records. Data were entered and analyzed by using statistical software SPSS version 20.0, and p < 0.05 was considered significant. RESULTS: The overall prevalence of hepatitis C among 708 HIV-infected patients was 130 (16.1 including 541 (76.4%) males and 167 (23.6%) females. High prevalence of HIV-HCV coinfection was significantly observed in males (122 [17.2%]) compared to females (8 [1.1%]) (p < 0.001). The main route of transmission among HIV-HCV coinfected patients was heterosexual contact (98 [13.8%]), followed by homosexual contact (4 [0.4%]). The statistically significant predictors involved in treatment outcomes of HIV-HCV coinfected patients are gender (OR = 2.015, p = 0.002) and intravenous drug users (OR = 2.376, p ≤ 0.001). CONCLUSION: The current study shows that HCV infection has an impact on the recovery of CD4 cells of the patients on HAART. Screening of HCV among HIV patients who were smokers and intravenous drug users should be monitored before starting HAART.