The RACK1 signal anchor protein from Trypanosoma brucei associates with eukaryotic elongation factor 1A: a role for translational control in cytokinesis

RACK1 is a WD-repeat protein that forms signal complexes at appropriate locations in the cell. RACK1 homologues are core components of ribosomes from yeast, plants and mammals. In contrast, a cryo-EM analysis of trypanosome ribosomes failed to detect RACK1, thus eliminating an important translational regulatory mechanism. Here we report that TbRACK1 from Trypanosoma brucei associates with eukaryotic translation elongation factor-1a (eEF1A) as determined by tandem MS of TAP-TbRACK1 affinity eluates, co-sedimentation in a sucrose gradient, and co-precipitation assays. Consistent with these observations, sucrose gradient purified 80S monosomes and translating polysomes each contained TbRACK1. When RNAi was used to deplete cells of TbRACK1, a shift in the polysome profile was observed, while the phosphorylation of a ribosomal protein increased. Under these conditions, cell growth became hypersensitive to the translational inhibitor anisomycin. The kinetoplasts and nuclei were misaligned in the postmitotic cells, resulting in partial cleavage furrow ingression during cytokinesis. Overall, these findings identify eEF1A as a novel TbRACK1 binding partner and establish TbRACK1 as a component of the trypanosome translational apparatus. The synergy between anisomycin and TbRACK1 RNAi suggests that continued translation is required for complete ingression of the cleavage furrow

Upper Gastrointestinal Bleeding Induced by Gastric Ulcer Secondary to Strongyloidiasis: A Case Report

Strongyloidiasis, a parasitic infestation by Strongyloides stercoralis, involves the gastrointestinal tract with a spectrum from duodenitis to enterocolitis. However, gastric involvement with the manifestation of upper gastrointestinal bleeding is an extremely rare condition due to Strongyloides stercoralis. Due to irregular excretion of larvae, unclear symptoms, paucity of effective diagnostic tools and low parasitic load, makes clinicians difficult to reach the diagnosis of strongyloidiasis. Here, we present a case of upper gastrointestinal bleeding due to a large gastric ulcer whose aetiology was identified to be Strongyloides stercoralis infection of the gastric region by the diagnosis of exclusion

Clinical significance of risk factor analysis in pancreatic cancer by using supervised model of machine learning

IntroductionPancreatic cancer (PC) poses a significant global health challenge due to its aggressive nature, late-stage diagnosis, and high mortality despite advancements in treatment. Early detection remains crucial for timely intervention. This study aimed to identify clinically relevant predictors of pancreatic cancer using a supervised machine learning approach and to develop a risk stratification tool with diagnostic capabilities.MethodsA matched case-control study was conducted retrospectively at the Tenth People’s Hospital of Tongji University (2017–2023), involving 353 cases and 370 matched controls. Demographic and hematological data were extracted from medical records. Variables were pre-selected using cluster dendrograms and subsequently refined using logistic regression with backward elimination and Support Vector Machine (SVM) models. A final risk scoring model was developed based on the best-performing model and internally validated.ResultsKey predictors retained in the final logistic regression model included Hemoglobin A1c (HbA1c) (OR 1.28; 95% CI: 1.08–1.52), Alkaline Phosphatase (ALP) (OR 1.02; 95% CI: 1.01–1.03), CA19-9 (OR 1.01; 95% CI: 1.01–1.01), Carcinoembryonic Antigen (CEA) (OR 1.41; 95% CI: 1.20–1.66), and Body Mass Index (BMI) (OR 0.88; 95% CI: 0.81–0.97). The final model demonstrated excellent diagnostic performance (AUC = 0.969, p < 0.001), with high accuracy, sensitivity, and specificity. A nomogram was constructed to facilitate individualized PC risk assessment.ConclusionHbA1c, ALP, CA19-9, CEA, and BMI were independently associated with pancreatic cancer. The machine learning-derived risk scoring model demonstrated high predictive accuracy and may serve as a valuable clinical tool for early detection and screening of pancreatic cancer

Transcript Expression Analysis of Putative Trypanosoma brucei GPI-Anchored Surface Proteins during Development in the Tsetse and Mammalian Hosts

Human African Trypanosomiasis is a devastating disease caused by the parasite Trypanosoma brucei. Trypanosomes live extracellularly in both the tsetse fly and the mammal. Trypanosome surface proteins can directly interact with the host environment, allowing parasites to effectively establish and maintain infections. Glycosylphosphatidylinositol (GPI) anchoring is a common posttranslational modification associated with eukaryotic surface proteins. In T. brucei, three GPI-anchored major surface proteins have been identified: variant surface glycoproteins (VSGs), procyclic acidic repetitive protein (PARP or procyclins), and brucei alanine rich proteins (BARP). The objective of this study was to select genes encoding predicted GPI-anchored proteins with unknown function(s) from the T. brucei genome and characterize the expression profile of a subset during cyclical development in the tsetse and mammalian hosts. An initial in silico screen of putative T. brucei proteins by Big PI algorithm identified 163 predicted GPI-anchored proteins, 106 of which had no known functions. Application of a second GPI-anchor prediction algorithm (FragAnchor), signal peptide and trans-membrane domain prediction software resulted in the identification of 25 putative hypothetical proteins. Eighty-one gene products with hypothetical functions were analyzed for stage-regulated expression using semi-quantitative RT-PCR. The expression of most of these genes were found to be upregulated in trypanosomes infecting tsetse salivary gland and proventriculus tissues, and 38% were specifically expressed only by parasites infecting salivary gland tissues. Transcripts for all of the genes specifically expressed in salivary glands were also detected in mammalian infective metacyclic trypomastigotes, suggesting a possible role for these putative proteins in invasion and/or establishment processes in the mammalian host. These results represent the first large-scale report of the differential expression of unknown genes encoding predicted T. brucei surface proteins during the complete developmental cycle. This knowledge may form the foundation for the development of future novel transmission blocking strategies against metacyclic parasites

Bullying Status on Secondary Level Students

Background: Bullying in school aged children is a universal problem, which continues to be a serious threat to the physical and emotional health of children and adolescents. The main objective of the study is to find out the status of bullying among secondary level school students in Kathmandu Nepal.Methods: A cross-sectional research design was carried in the public and private level secondary school of Kathmandu in 2018. The cross sectional descriptive analytical study design along with the case study method was used among 222 respondents who were selected by purposive sampling. A semi structured questionnaire for socio-demographic and a structured of Olweus Bully Questionnaire (OBQ) was used for data collection. The data analysis was done using SPSS version 21.Result: Among 222, total 146 (65.8%) of the students got bullied physically, verbally and sexually. Only (14.4%) of the students bullied by electronic devices or cyberbullying. However, the number of students involved in bullying others is 100 (45%) only. The number of boys who got bullied is 176 (79.28%) which is higher than that of girls 144 (64.85%). Majority 45.63% of the bullying occurred in the classroom when the teacher was not present and bullying occurred least on the way to home, school bus stop, and gym class. Mostly 50% report their problem to the friends followed by brother or sister, class teacher, parents, another adult at school, somebody else 7.89%, 18.42%, 10%,53%, 6.58 %, 6.58% respectively. The findings show that there is no association between the socio-demographic variables and bullying among the secondary level student’s in Kathmandu Nepal.Conclusion: The study found that the status of bullying highly prevails in the schools of Kathmandu, Nepal. Hence, all stakeholders in secondary education should work collaboratively for interventions to reduce the bulling practice and its adverse effects on student health.

Immune Checkpoint Inhibitor (ICI)-Induced Painful Thyroiditis: A Rare Presentation With Thyrotoxicosis

Description: Thyroid dysfunction occurs in 5% to 15% of patients undergoing Immune Checkpoint Inhibitor (ICI) therapy, such as pembrolizumab, typically presenting as painless thyroiditis followed by hypothyroidism. Although often mild and manageable, rare cases of painful thyroiditis with overt thyrotoxicosis can arise, potentially complicating treatment. Case Report: A 23-year-old female with breast cancer treated with pembrolizumab presented with 6 days of fever, chills, erythema, and tenderness at a port site. Despite port removal and antibiotics, fevers persisted with tachycardia. An infectious workup and repeat were negative. Thyrotoxicosis was identified; TSH \u3c 0.008 mIU/L (0.35-4.94), free T4 3.23 ng/dL (0.70-1.48); thyroid tenderness and elevated ESR, consistent with thyroiditis. Free T3 was normal. Thyroid ultrasound showed diffuse gland enlargement. Initial treatment with prednisone, methimazole, and propranolol improved symptoms. After a contrast CT, fever and odynophagia worsened, managed with IV dexamethasone and increased methimazole. At discharge, she remained thyrotoxic; however, fever subsided. Endocrinology follow-up was arranged. Discussion: ICI-mediated thyroiditis, seen in cancer patients undergoing immunotherapy, typically begins with a thyrotoxic phase, often asymptomatic, and transitions rapidly to likely permanent hypothyroidism. Unlike other thyroiditis etiologies, it is not included in hyperthyroidism or thyrotoxicosis guidelines, necessitating a tailored approach. Painless thyroiditis from ICIs usually involves transient thyrotoxicosis in 5% to 20% of cases, followed by hypothyroidism lasting about 6 months, with most recovering normal function, though 10% to 20% develop permanent hypothyroidism. Unlike subacute or postpartum thyroiditis, ICI-mediated thyroiditis is painless, transitions faster (3 months vs 6-9 months), and may involve elevated TPO or thyroglobulin antibodies. Conclusion: This case emphasizes the importance of recognizing atypical patterns in ICI-mediated thyroiditis to ensure timely and appropriate intervention. The requirement for high-dose IV steroids and escalating thionamide doses highlights the severity of the condition, deviating from the usual mild and transient course of ICI-related thyroid dysfunction. This further underscores the need for close monitoring of thyroid function in patients undergoing immunotherapy to prevent complications such as thyroid storm. Guidelines recommend checking thyroid function tests every 4-6 weeks from therapy initiation and every 2-3 weeks after a thyroiditis diagnosis to detect hypothyroid conversion

EFFICACY OF COMMERCIAL INSECTICIDES AGAINST FRUIT AND SHOOT BORER, LEUCINODES ORBONALIS GUENEE OF BRINJAL, SOLANUM MELONGENA L. IN DHADING, NEPAL

A study was conducted from January to June 2019 in Dhading district to identify the effective commercial insecticide against fruit and shoot borer of brinjal (Leucinodes orbonalis). A field experiment was conducted in Randomized Completely Block Design with five treatments (T1: Spinosad @ 1 ml/ 3 liters, T2: Chlorantraniliprole 1 ml/ 3 liters, T3: Cypermethrin 3 ml/ 1 liter, T4: Neem 3 ml/ 1 liter and T5: Control) and four replications. Result indicated that the fruit infestation percent and shoot damage reduction percent, fruit infestation reduction percent were highly significant with the application of insecticides. The shoot damage percent and fruit damage percent was the lowest with spinosad followed by chlorantraniliprole, cypermethrin and Neem. Thus, spinosad is the most effective insecticides for the management of fruit and shoot borer of brinjal in field conditions.</jats:p

Translational initiation in Leishmania tarentolae and Phytomonas serpens (Kinetoplastida) is strongly influenced by pre-ATG triplet and its 5' sequence context

To investigate the influence of sequence context of translation initiation codon on translation efficiency in Kinetoplastida, we constructed a library of expression plasmids randomized in the three nucleotides prefacing ATG of a reporter gene encoding enhanced green fluorescent protein (EGFP). All 64 possible combinations of pre-ATG triplets were individually stably integrated into the rDNA locus of Leishmania tarentolae and the resulting cell lines were assessed for EGFP expression. The expression levels were quantified directly by measuring the fluorescence of EGFP protein in living cells and confirmed by Western blotting. We observed a strong influence of the pre-ATG triplet on the level of protein expression over a 20-fold range. To understand the degree of evolutionary conservation of the observed effect, we transformed Phytomonas serpens, a trypanosomatid parasite of plants, with a subset of the constructs. The pattern of translational efficiency mediated by individual pre-ATG triplets in this species was similar to that observed in L. tarentolae. However, the pattern of translational efficiency of two other proteins (red fluorescent protein and tetracycline repressor) containing selected pre-ATG triplets did not correlate with either EGFP or each other. Thus, we conclude that a conserved mechanism of translation initiation site selection exists in kinetoplastids that is strongly influenced not only by the pre-ATG sequences but also by the coding region of the gene

Upper Gastrointestinal Bleeding Induced by Gastric Ulcer Secondary to Strongyloidiasis: A Case Report

Strongyloidiasis, a parasitic infestation by Strongyloides stercoralis, involves the gastrointestinal tract with a spectrum from duodenitis to enterocolitis. However, gastric involvement with the manifestation of upper gastrointestinal bleeding is an extremely rare condition due to Strongyloides stercoralis. Due to irregular excretion of larvae, unclear symptoms, paucity of effective diagnostic tools and low parasitic load, makes clinicians difficult to reach the diagnosis of strongyloidiasis. Here, we present a case of upper gastrointestinal bleeding due to a large gastric ulcer whose aetiology was identified to be Strongyloides stercoralis infection of the gastric region by the diagnosis of exclusion.</jats:p

Salivary gland specific gene expression in metacyclic trypanosomes in saliva.

+<p> = transcript detected.</p><p>Eight genes previously determined to be specifically expressed in salivary glands were analyzed from metacyclic cDNAs by RT-PCR analysis.</p