Assessment of menisci and ligamentous injuries of the knee on magnetic resonance imaging: correlation with arthroscopy

OBJECTIVE: To evaluate the validity of MRI, in the assessment of the meniscal and cruciate ligamenteous injuries of the knee joint and comparison with arthroscopic findings. METHODS: A one year prospective cross-sectional study from January 2006 to January 2007, was performed on 50 patients (32 males & 18 females) with knee injury presenting at the orthopedic unit of AKUH. The patients were referred to radiology department for MRI evaluation and arthroscopy. RESULTS: The sensitivity, specificity and accuracy for MRI of the menisci and ligaments were as follows: medial meniscus resulted in 100% sensitivity, 69.27% specificity, 90% PPV, 100% NPV and 92% accuracy: lateral meniscus resulted in 87.5% sensitivity, 88.23% specificity, 77% PPV, 93% NPV and 88% accuracy: anterior cruciate ligament resulted in 86.67% sensitivity, 91.43% specificity, 81% PPV, 94% NPV and 88% accuracy; posterior cruciate ligament resulted in 100% sensitivity, 95.83% specificity,50% PPV, 100 NPV and 96% accuracy. CONCLUSION: Magnetic resonance imaging is a good, accurate and non invasive modality for the assessment of menisci and ligamenteous injuries. It can be used as a first line investigation in patients with soft tissue trauma to knee

Effects of Salinity Stress on Growth and Physio-biochemical Parameters of Three Pea (Pisum sativum L.) Cultivars of Different Maturity Duration

Background: Salinity is one of the leading abiotic stresses that negatively affects the growth of many important food crops and significantly reduces the productivity and yield value.Methods: The present study was conducted to study the effects of NaCl stress on three pea (Pisum sativum) cultivars (Climax, Lina Pak and Pea-267) of different maturity level (Late, early, and mid-season flowering) under In vitro conditions. Two weeks old In vitro grown shoots of three pea cultivars were subjected to stress condition in MS medium supplemented with five levels of NaCl (0, 20, 40, 80 and 100mM NaCl) for one month and different morphological and physio-biochemical traits including length of shoot, number of leaves, shoot biomass, chlorophyll, proline and total phenolic content, total proteins and non-enzymatic antioxidant (DPPH) activities were studied.Results: The results were analyzed using different statistical approaches (ANOVA, MNOVA, PCA, correlation and regression) to identify the tolerance level of each genotype. Shoot length and shoot fresh weight were increased at 20 and 40mM in Climax, while proline content progressively increased with an increase in stress concentration in all the genotypes. Total protein content increased in cvs. Climax and Pea-267 and  decreased in Lina Pak above 20mM and DPPH was increased in Climax and Pea-267 at 20 and 40mM, while in Lina Pak it showed an increase at only 20mM NaCl concentration. According to the results of MNOVA and regression analysis, significant changes occurred in biomass, proline content and DPPH values. A strong positive correlation of shoot dry weight was found with total phenolic and proline content. Maximum value of stress tolerance index was recorded for Climax.Conclusion: Biplot analysis clustered cvs. Climax and Pea-267 cultivars into tolerant group and Lina Pak in sensitive group based on the mean performance of studied parameters to NaCl stress and control treatments

Enteric Fever as an Antecedent to Development of Miller-Fisher Syndrome and Possible Role of COVID-19 Vaccination

Summary: Guillain-Barre Syndrome is an immune-mediated demyelinating disorder. Miller-Fisher Syndrome is an uncommon subtype of GBS. It is characterized by findings of ophthalmoplegia, ataxia, and areflexia. Here we present the case of Miller-Fisher Syndrome following an episode of typhoidal diarrhea. The presentation was of rapidly progressing weakness beginning in the lower extremity with diplopia. Examination revealed diminished reflexes. CSF testing revealed albuminocytologic dissociation which was later supported by neurophysiological testing. The patient was treated with intravenous immunoglobulins (IVIG). We conclude that Miller-Fisher syndrome should be considered in the diagnostic workup of patients presenting with new sensorimotor deficits following diarrheal illnesses and/or COVID-19 mRNA vaccination. Early recognition is essential given the propensity of GBS to cause life-threatening respiratory failure and prompt IVIG administration is associated with a better prognosis. Keywords: Enteric Fever, Miller-Fisher Syndrome, COVID-19, Vaccinatio

Androgens upregulate Cdc25C protein by inhibiting its proteasomal and lysosomal degradation pathways.

Cdc25C is a cell cycle protein of the dual specificity phosphatase family essential for activating the cdk1/Cyclin B1 complex in cells entering into mitosis. Since altered cell cycle is a hallmark of human cancers, we investigated androgen regulation of Cdc25C protein in human prostate cancer (PCa) cells, including androgen-sensitive (AS) LNCaP C-33 cells and androgen-independent (AI) LNCaP C-81 as well as PC-3 cells. In the regular culture condition containing fetal bovine serum (FBS), Cdc25C protein levels were similar in these PCa cells. In a steroid-reduced condition, Cdc25C protein was greatly decreased in AS C-33 cells but not AI C-81 or PC-3 cells. In androgen-treated C-33 cells, the Cdc25C protein level was greatly elevated, following a dose- and a time-dependent manner, correlating with increased cell proliferation. This androgen effect was blocked by Casodex, an androgen receptor blocker. Nevertheless, epidermal growth factor (EGF), a growth stimulator of PCa cells, could only increase Cdc25C protein level by about 1.5-fold. Altered expression of Cdc25C in C-33 cells and PC-3 cells by cDNA and/or shRNA transfection is associated with the corresponding changes of cell growth and Cyclin B1 protein level. Actinomycin D and cycloheximide could only partially block androgen-induced Cdc25C protein level. Treatments with both proteasomal and lysosomal inhibitors resulted in elevated Cdc25C protein levels. Immunoprecipitation revealed that androgens reduced the ubiquitination of Cdc25C proteins. These results show for the first time that Cdc25C protein plays a role in regulating PCa cell growth, and androgen treatments, but not EGF, greatly increase Cdc25C protein levels in AS PCa cells, which is in part by decreasing its degradation. These results can lead to advanced PCa therapy via up-regulating the degradation pathways of Cdc25C protein

Molecular epidemiology of SARS-CoV-2: A tertiary care hospital experience from Pakistan

Objectives: The current study was conducted to assess the molecular epidemiology of SARS-CoV-2 in the general population. Methodology: This study was conducted from April to July 2020, at the Molecular Diagnostic Laboratory, Pakistan Atomic Energy Commission (PAEC) General Hospital Islamabad, Pakistan. A total of 28,274 nasopharyngeal swabs were collected in Viral Transport Medium (VTM) media from symptomatic and asymptomatic individuals at the sample collection centers of our hospital and other affiliated hospitals. RNA was extracted using both automated and manual extraction platforms as per the manufacturer's instructions. Multiple qualitative reverse transcription real-time PCR kits for the identification of SARS-CoV-2 were used. Results: The results showed that 1,722 (6.09%) were positive for SARA-CoV-2 RNA. The males exhibited a prevalence of 2.76% while females showed a high prevalence of 13.44%. Among males,  most patients 424 (31.47%) were in the age group of 31-40 years followed by the age group of 41-50 years 306 (22.71%). Similarly among females, the majority of patients were from the age group 31-40 years with 91 (24.66%) followed by 41-50 years of age group 70 (18.66%) confirmed cases. Conclusion: The molecular epidemiological data may support the national policy formulation, transmission tracking, and the execution of measures to control viral transmission

Huh-7 cell line as an alternative cultural model for the production of human like erythropoietin (EPO)

Erythropoietin (EPO) is a glycoprotein hormone which is required to regulate the production of red blood cells. Deficiency of EPO is known to cause anemia in chronically infected renal patients and they require regular blood transfusion. Availability of recombinant EPO has eliminated the need for blood transfusion and now it is extensively used for the treatment of anemia. Glycosylation of erythropoietin is essential for its secretion, stability, protein conformation and biological activity. However, maintenance of human like glycosylation pattern during manufacturing of EPO is a major challenge in biotechnology. Currently, Chinese hamster ovary (CHO) cell line is used for the commercial production of erythropoietin but this cell line does not maintain glycosylation resembling human system. With the trend to eliminate non-human constituent from biopharmaceutical products, as a preliminary approach, we have investigated the potential of human hepatoma cell line (Huh-7) to produce recombinant EPO.Initially, the secretory signal and Kozak sequences was added before the EPO mature protein sequence using overlap extension PCR technique. PCR-amplified cDNA fragments of EPO was inserted into mammalian expression vector under the control of the cytomegalovirus (CMV) promoter and transiently expressed in CHO and Huh-7 cell lines. After RT-PCR analysis, ELISA and Western blotting was performed to verify the immunochemical properties of secreted EPO.Addition of secretory signal and Kozak sequence facilitated the extra-cellular secretion and enhanced the expression of EPO protein. Significant expression (P < 0.05) of EPO was observed in the medium from Huh-7 cell line.Huh-7 cell line has a great potential to produce glycosylated EPO, suggesting the use of this cell line to produce glycoproteins of the therapeutic importance resembling to the natural human system

Phytochemicals and bioactivities of Syzygium filiforme var. filiforme

Syzygium filiforme var. filiforme is a plant variety from dicotyledonous plant family (Myrtaceae). Phytochemical studies on S. filiforme var. filiforme stem bark have successfully isolated and characterized arjunolic acid (1), alphitolic acid (2), betulinic acid (3), ursolic acid (4), ursolic acid 3-methyl ester (5), β-sitosterol (6) and stigmasterol (7). The inhibitory activities against free radical, starch, and bacteria for major compounds were tested by using DPPH, α-glucosidase and minimum inhibitory and bacterial concentration assays, respectively. No promising antioxidant activity was shown on tested samples except methanolic crude extract. For antidiabetic activity, methanolic and dichloromethane crude extracts displayed potent activity compared to 1-deoxynojirimycin. Minimum inhibitory concentration (MIC) and minimum bacterial concentration (MBC) assays for antibacterial activity were evaluated on Escherichia coli, Staphylococcus aureus, and Bacillus subtilis. All crude extracts and major compounds displayed weak and no promising activities for MIC method, respectively. Meanwhile, for MBC method, hexane crude extract and compound 1 showed inhibition against B. subtilis

HCV genotype-specific correlation with serum markers: Higher predictability for genotype 4a

Several factors have been proposed to assess the clinical outcome of HCV infection. The correlation of HCV genotypes to possible serum markers in clinical prediction is still controversial. The main objective of this study was to determine the existence of any correlation between HCV genotypes to viral load and different clinical serum markers.We performed a prospective cross-sectional and observational study. About 3160 serum HCV RNA positive patients were chosen from 4020 randomly selected anti-HCV positive patients. Statistical analysis was performed using the SPSS 16 software package. ROC (receiver operating characteristics) curves were used to compare diagnostic values of serum markers to predict genotypes.The most prevalent genotype was 3a (73.9%) followed by 1a (10.7%), 4a (6.4%) and 3b (6.1%) in Pakistani population. No correlation was found between viral load and serum markers for genotype 3a in a large no. of sample (n = 2336). While significant correlation was observed between viral load and AST in genotype 3b, ALP with viral load and ALT for genotype 1a. Patients with genotype 4a showed a significant inverse correlation with viral load and Hb level and AST with ALP. For genotype 4a, AUC (area under the curve) of ALT, ALP, AST, bilirubin, Hb level and viral load was 0.790, 0.763, 0.454, 0.664, 0.458 and 0.872 respectively.In conclusion, there was a significant variable response of HCV genotypes with serum markers. Severity of disease is independent of serum marker level in genotype 3a, while the liver damage in genotype 4a may associate with viral cytopathic effect as well as the immune-mediated process. An index using six serum markers may correctly predict genotype 4a in patients with ≥ 75% accuracy

Claudin-1 required for HCV virus entry has high potential for phosphorylation and O-glycosylation

HCV is a leading cause of hepatocellular carcinoma and cirrhosis all over the world. Claudins belong to family of tight junction's proteins that are responsible for establishing barriers for controlling the flow of molecules around cells. For therapeutic strategies, regulation of viral entry into the host cells holds a lot of promise. During HCV infection claudin-1 is highly expressed in liver and believed to be associated with HCV virus entry after HCV binding with or without co-receptor CD81. The claudin-1 assembly with tight junctions is regulated by post translational modifications. During claudins assembly and disassembly with tight junctions, phosphorylation is required at C-terminal tail. In cellular proteins, interplay between phosphorylation and O-β-GlcNAc modification is believed to be functional switch, but it is very difficult to monitor these functional and vibrant changes in vivo. Netphos 2.0 and Disphos 1.3 programs were used for potential phosphorylation; NetPhosK 1.0 and KinasePhos for kinase prediction; and YinOYang 1.2 and OGPET to predict possible O-glycosylation sites. We also identified Yin Yang sites that may have potential for O-β-GlcNAc and phosphorylation interplay at same Ser/Thr residues. We for the first time proposed that alternate phosphorylation and O-β-GlcNAc modification on Ser 192, Ser 205, Ser 206; and Thr 191 may provide an on/off switch to regulate assembly of claudin-1 at tight junctions. In addition these phosphorylation sites may be targeted by novel chemotherapeutic agents to prevent phosphorylation lead by HCV viral entry complex

Toll-Like Receptor 7 Stimulates the Expression of Epstein-Barr Virus Latent Membrane Protein 1

Epstein-Barr virus (EBV) is a ubiquitous human herpesvirus. Toll-like receptor 7 (TLR7) is involved in host innate immunity against pathogens, and its aberrant activation is linked to the development of systemic lupus erythematosus (SLE, also called ‘‘lupus’’). Type I interferons (IFN) are apparently driving forces for lupus pathogenesis. Previously, we found that EBV latent membrane protein 1 (LMP1) primes cells for IFN production. In this report, the relationship among EBV LMP1, TLRs, and IFN production are examined. We find that TLR7 activation increases the expression of EBV LMP1, and IFN regulatory factor 7 (IRF7) is involved in the stimulation process. TLR7 activation did not induce IFNs from EBV-infected cells, but potentiates those cells for IFN production by TLR3 or TLR9 activation. In addition, we find that LMP1 and IFNs are coexpressed in the same cells in some lupus patients. Therefore, the aberrant activation of TLR7 might induce LMP1 expression and LMP1-expression cells may be producing IFNs in lupus patients. These results suggest EBV might be an exacerbating factor in some lupus patients via promoting IFN production