CD8α-CI-M6PR Particle Motility Assay to Study the Retrograde Motion of CI-M6PR Receptors in Cultured Living Cells.

The cation-independent mannose 6-phosphate receptors (CI-M6PR) bind newly synthesized mannose 6-phosphate (Man-6-P)-tagged enzymes in the Golgi and transport them to late endosomes/lysosomes, providing them with degradative functions. Following the cargo delivery, empty receptors are recycled via early/recycling endosomes back to the trans-Golgi network (TGN) retrogradely in a dynein-dependent motion. One of the most widely used methods for studying the retrograde trafficking of CI-M6PR involves employing the CD8α-CI-M6PR chimera. This chimera, comprising a CD8 ectodomain fused with the cytoplasmic tail of the CI-M6PR receptor, allows for labeling at the plasma membrane, followed by trafficking only in a retrograde direction. Previous studies utilizing the CD8α-CI-M6PR chimera have focused mainly on colocalization studies with various endocytic markers under steady-state conditions. This protocol extends the application of the CD8α-CI-M6PR chimera to live cell imaging, followed by a quantitative analysis of its motion towards the Golgi. Additionally, we present an approach to quantify parameters such as speed and track lengths associated with the motility of CD8α-CI-M6PR endosomes using the Fiji plugin TrackMate. Key features • This assay is adapted from the methodology by Prof. Matthew Seaman for studying the retrograde trafficking of CI-M6PR by expressing CD8α-CI-M6PR chimera in HeLa cells. • The experiments include live-cell imaging of surface-labeled CD8α-CI-M6PR molecules, followed by a chase in cells. • Allows the monitoring of real-time motion of CD8α-CI-M6PR endosomes and facilitates calculation of kinetic parameters associated with endosome trajectories, e.g., speed and distance (run lengths).

An epidemiological study of the determinants and patterns of complementary feeding practices among children of 6-24 months of age group.

Background: The introduction of complementary food is the most important factor determining the nutritional status of the children in the first 2 years of life. This study aimed to find out the determinants and patterns of complementary feeding practices and their impact on the growth and development of children of 6-24 months of age group. Materials and Methods: A descriptive cross-sectional study was done on immunization out patient department (OPD) in the field practice area of the Department of Community Medicine. The sample size taken was 250 using a systematic random sampling method. Results: One hundred and seven (42.8%) were given the optimum amount of complementary food for their age. One hundred and three (41.2%) were fed with an adequate frequency for their age. One hundred and ninety-five (78.0%) were given food that was appropriate in consistency for their age. Among 183 (73.2%) children who were not fed appropriately during and after their illness episodes, 119 (65.0%) were malnourished. On using logistic regression it was seen that the variables which had statistically significant association with the complementary feeding patterns (P<0.05) were maternal education, father's age, total members in the family, advice on complementary feeding from health care workers and use of bottle for feeding. Father's education, availability of ration card, antenatal counselling on childcare, birth order and history of repeated illness in the past did not show statistically significant association with complementary feeding practices (P>0.05). Conclusion: There was no statistically significant difference between the feeding patterns of normal and malnourished children. The association of complementary feeding practices and the anthropometric status of study children was not found statistically significant in the present study. Advice on complementary feeding from healthcare workers is seen to be having a positive impact on complementary feeding practices (P < 0.01).

Awareness and utilization of mother and child protection (MCP) card among mothers attending immunization OPD: An urban health centre-based cross-sectional study.

Background: The MCP card is used as a tool for pregnant women, young mothers and family members to learn about various types of services which they should assess and utilize. The study was undertaken to see the awareness about MCP card and its components among beneficiaries in urban slum area. Methodology: This is a cross-sectional observational study done in urban slum immunization OPD with a sample size of 226. Convenient sampling method was used to select the samples. Mothers consenting and willing to participate in the study were enrolled. Results: Observed that the MCP card was issued to majority participants at the time of baby immunization (44.15%), followed by at time of delivery (31.4%) and least number received it at the time of pregnancy registration (24.34%). It is also observed that only 53.54%(121) women had read the card while the remaining didn't. Majority thought the primary use of MCP card was vaccination record (91.5%), followed by information tool (52.%). It is observed that knowledge regarding pregnancy care and its related complications leads to better utilization of ANC services. Conclusion: As observed in the present study a sizeable proportion of populations doesn't even read the card, and among those who read it the level of knowledge isn't satisfactory, many use the card as a vaccination record but still the awareness about various vaccines among the beneficiaries remains low.

A baseline cross-sectional study to assess patient satisfaction (18-60 years) toward Apli Chikitsa (free of cost medical laboratory tests) services provided at an urban health training center: A PPP model.

Introduction: Assessing satisfaction with healthcare services is one of the cornerstone methods to evaluate success and identify obstacles during the process for the patients. Identifying crucial information helps in course correction and better healthcare delivery to the patients. Apli Chikitsa is a novel PPP model-based free of cost laboratory service made available to the patients by the municipal corporation. Around 150 blood tests are offered free of cost and through peripheral dispensaries to the patients. The present study was undertaken to assess the satisfaction of patients with the service and service-related aspects. Methodology: A cross-sectional observational urban health center-based study was undertaken to assess the satisfaction of patients toward the Apli Chikitsa laboratory service. Institutional ethics committee clearance was obtained before commencing the study. Four hundred patients availing of the laboratory services were enrolled by alternate sampling method. A semistructured questionnaire was administered; five-point Likert scale was used to assess satisfaction toward the service and related aspects. Results were summarized using tables and charts, and percentages and proportions were used wherever necessary. Data were analyzed using Microsoft Excel. Results: Among the study participants, overall satisfaction was observed among 71.25%, and 100% satisfaction was observed in relation to cost of service, maintenance of privacy and confidentiality, and getting all the requested tests done. The problem of overcrowding (96.75%) was observed by a majority. Satisfaction was low regarding the cleanliness of toilets (48.75%). Conclusion: Based on the findings, it can be concluded that the overall satisfaction is the Apli Chikitsa laboratory service is good, and it is a good model for replication for laboratory service delivery in peripheral areas away from tertiary care centers. Overcrowding should be managed, and cleanliness of toilets to done at frequent intervals, along with proper and visible use of signages, should be done to improve the satisfaction of the patients.

A cross sectional study of determinants of bone mineral density among postmenopausal women with special reference to anthropometric and lifestyle factors in an urban slum of Mumbai.

Background and Aim: Osteoporosis significantly compromises the quality of life of postmenopausal women. There are DALY (Disability-Adjusted Life Years) losses for postmenopausal women who suffer from osteoporosis. Therefore, this study was taken up with the objective to determine prevalence of osteopenia and osteoporosis among postmenopausal women and to study the association of their anthropometric parameters and lifestyle factors with their bone mineral density (BMD). To study the determinants of BMD among postmenopausal women with special reference to anthropometric and lifestyle factors in an urban slum. Materials and Methods: A community-based, cross-sectional study was conducted in an urban slum of Mumbai. The study was conducted among 148 postmenopausal women. BMD of the study group was measured using portable, noninvasive Calcaneum Ultrasound Bone Densitometer, and T-score reading was noted. Lifestyle factors and dietary habits were assessed through a prestructured, validated questionnaire. Body mass index (BMI) was calculated with a standard procedure. No trial was conducted in the study. Statistical Package for Social Sciences (version 20) was used for statistical analysis. Results: The prevalence of osteopenia among study participants was found to be 43.9% and that of osteoporosis was 12.8%. Their BMI was in the range of 16.44-39.18 kg/m2. Walking, stretching exercises, and yoga were the form of exercise being practiced by 29.1% of postmenopausal women. Conclusions: BMD was significantly associated with age, socioeconomic status, BMI, exercise, walking, and consumption of fruits. Negative correlation was found between duration since menopause and BMD T-score reading, which implied that duration since menopause had an inverse relationship with BMD.

Bone marrow metastasis in nonhematological malignancies: A study from tertiary care center.

Introduction: Metastatic cancer presents a treatment challenge to clinicians, particularly for patients with bone marrow infiltration. For tumor staging, therapy selection, and prognosis risk stratification, the status of the bone marrow should be known for the presence or absence of metastasis. The study aimed to evaluate the hematological findings and comprehensive analysis of bone marrow in cases of nonhematological malignancies with bone marrow metastasis. Materials and Methods: This retrospective study comprised a record retrieval of the departmental archives for the past 6 years. A total of 331 patients with nonhematological malignancies were found, of whom 31.42% (104/331) showed bone marrow metastasis. An integrated clinical approach with bone marrow examination findings and immunohistochemistry whenever necessary was used to achieve a definitive diagnosis of bone marrow metastasis. Results: Among the study population, 31.42% (104/331) of patients had nonhematological malignancies that metastasized to the bone marrow. Most of the patients with bone marrow metastasis had anemia, which was found in 77.88% (81/104) of the cases. Leukoerythroblastic reaction was noted in 31.73% (33/104) of the cases, and thrombocytopenia was found in 25% (26/104) of the cases. The most common malignancy with bone marrow metastasis in adults was prostatic adenocarcinoma (28.1%) (9/32) and in pediatric cases, neuroblastoma (53.9%) (52/98). Conclusions: It is essential to diagnose nonhematological malignancies that have metastasized to the bone marrow since this necessitates tumor staging, therapy selection, and prognosis risk stratification. To conclude, not a single hematological parameter is predictive of bone marrow metastasis; however, unexplained anemia, a leukoerythroblastic blood picture, and thrombocytopenia in peripheral blood should raise suspicion for bone marrow metastasis in cases of nonhematological malignancies.

Résumé Introduction: Le cancer métastatique présente un défi de traitement pour les cliniciens, en particulier pour les patients présentant une infiltration de moelle osseuse. Pour la stadification tumorale, la sélection du traitement et la stratification du risque de pronostic, l'état de la moelle osseuse doit être connu pour la présence ou l'absence de métastases. L'étude visait à évaluer les résultats hématologiques et l'analyse complète de la moelle osseuse dans les cas de tumeurs malignes non hématologiques avec métastases de la moelle osseuse. Matériel et méthodes: Cette étude rétrospective comprenait une récupération des archives ministérielles des 6 dernières années. Un total de patients atteints de tumeurs malignes non hématologiques ont été trouvés, dont 31,42% (104/331) présentaient des osmétastases médullaires. Une approche clinique intégrée avec les résultats de l'examen de la moelle osseuse et l'immunohistochimie chaque fois que nécessairea été utilisé pour établir un diagnostic définitif de métastases médullaires. Résultats: Dans la population étudiée, 31,42 % (104/331) des patients présentaient des tumeurs malignes non hématologiques qui se métastasaient à la moelle osseuse. La plupart des patients atteints de métastases de la moelle osseuse présentaient une anémie, qui a été trouvée dans 77,88% (81/104) des cas. Une réaction leucoérythroblastique a été observée dans 31,73 % (33/104) des cas, et une thrombocytopénie a été observée dans 25 % (26/104) des cas. La tumeur maligne la plus fréquente associée aux métastases de la moelle osseuse chez l'adulte était l'adénocarcinome de la prostate (28,1 %) (9/32) et, chez les enfants, le neuroblastome (53,9 %) (52/98). Conclusions: Il est essentiel de diagnostiquer les tumeurs malignes non hématologiques qui ontmétastasé à la moelle osseuse car cela nécessite une stadification tumorale, une sélection thérapeutique et une stratification du risque de pronostic. Pour conclure, pas un seul paramètre hématologique n'est prédictif des métastases de la moelle osseuse; Cependant, une anémie inexpliquée, une image sanguine leucoérythroblastique et une thrombocytopénie dans le sang périphérique devraient faire suspecter des métastases de la moelle osseuse en cas de tumeurs malignes non hématologiques. Mots-clés: Aspiration de moelle osseuse, biopsie de la moelle osseuse, métastases de la moelle osseuse, résultats hématologiques, immunohistochimie, tumeurs malignes non hématologiques, frottis sanguin périphérique.