A study on diagnostic performance of different immunohematological diagnostic tests in assessing the prevalence of ABO Hemolytic Disease of Newborn in the antenatal O group mothers and their neonatal outcome in a tertiary care hospital in Northern India.

BACKGROUND: Hemolytic disease of the newborn (HDN) results in the decreased lifespan of the red cells. HDN related to ABO incompatibility is mostly unnoticed because routine screening is not being done. This study was done to assess the prevalence of ABO-HDN and to compare different immunohematological tests. Methods-In this study 213 O group mothers and the 122 ABO-incompatible newborns born to them were included. Quantifying the maternal IgG anti-A/anti-B antibody titer was done by Conventional Tube Technique (CTT) using Dithiothreitol (DTT) pretreated maternal serum. Hemolysin test was performed on the mothers having titer > 256. These cases were followed up and, after delivery, were monitored for ABO HDN, along with direct antiglobulin testing and elution studies. The prevalence of ABO-HDN was calculated, and the different diagnostic parameters of the tests were calculated. Results- The prevalence of ABO-HDN in our population was estimated to be 1.7%, 6.1% & 10.6% in our population, O group mothers, and O group mothers with ABOincompatible newborns, respectively. Maternal titer≥ 512 strongly correlated with ABOHDN. DAT positivity is a good predictor of ABO-HDN, especially using sensitive techniques. Maternal IgG titers have the highest sensitivity & Negative Predictive Value, while DAT has the highest specificity & Positive Predictive Value. Conclusion - Maternal ABO antibody titration may be advocated in the centers to identify high-risk groups. It can advocate institutional delivery and dedicated follow-up of newborns with ABO-HDN. Blood grouping & DAT may be performed in all newborns born to O blood group to identify high-risk cases.

Off with the blues: Therapeutic plasma exchange in a case of copper sulphate poisoning.

CuSO4 (Copper sulphate) poisoning though rare, is associated with high mortality. It involves multiple organ systems and if not dealt with promptly can lead to death. Supportive care and chelation therapy along with TPE (therapeutic plasma exchange), whole blood exchange or red cell exchange can be employed in management. We report such a case where swift clinical improvement was seen after TPE.

Binding of DNA origami to lipids: maximizing yield and switching via strand displacement.

Liposomes are widely used as synthetic analogues of cell membranes and for drug delivery. Lipid-binding DNA nanostructures can modify the shape, porosity and reactivity of liposomes, mediated by cholesterol modifications. DNA nanostructures can also be designed to switch conformations by DNA strand displacement. However, the optimal conditions to facilitate stable, high-yield DNA-lipid binding while allowing controlled switching by strand displacement are not known. Here, we characterized the effect of cholesterol arrangement, DNA structure, buffer and lipid composition on DNA-lipid binding and strand displacement. We observed that binding was inhibited below pH 4, and above 200 mM NaCl or 40 mM MgCl2, was independent of lipid type, and increased with membrane cholesterol content. For simple motifs, binding yield was slightly higher for double-stranded DNA than single-stranded DNA. For larger DNA origami tiles, four to eight cholesterol modifications were optimal, while edge positions and longer spacers increased yield of lipid binding. Strand displacement achieved controlled removal of DNA tiles from membranes, but was inhibited by overhang domains, which are used to prevent cholesterol aggregation. These findings provide design guidelines for integrating strand displacement switching with lipid-binding DNA nanostructures. This paves the way for achieving dynamic control of membrane morphology, enabling broader applications in nanomedicine and biophysics.

Irregular erythrocyte antibodies among antenatal women and their neonatal outcome at a tertiary care hospital in Northern India.

BACKGROUND: Red blood cell alloimmunisation during the pregnancy is a significant cause for neonatal mortality and morbidity. This study was planned to determine the prevalence and specificity of irregular erythrocyte antibodies in antenatal mothers and their neonatal outcome. METHODS: In this observational study, blood grouping and red cell antibody screening of mothers were performed at first visit and after 28 weeks of gestation and positive cases were identified and followed up monthly till delivery by repeating antibody titre and middle cerebral artery-peak systolic velocity. After delivery of alloimmunised mothers, cord blood haemoglobin, bilirubin and direct antiglobulin tests (DAT) were analysed and further outcome of neonate was recorded. RESULTS: Among 652 registered antenatal cases, 18 multigravida women were found to be alloimmunised, accounting to prevalence of 2.8%. Most common alloantibody identified was anti D (>70%) followed by anti-Lea, anti-C, anti-Leb, anti-E and anti-Jka. Only 47.7% Rh D negative women received anti-D prophylaxis during previous pregnancies or whenever indicated. DAT was positive in 56.2% of neonates. Among nine DAT positive neonates, two early neonatal deaths due to severe anaemia were observed following birth resuscitation. Four antenatal mothers required intrauterine transfusion in view of fetal anaemia while three neonates received double volume exchange transfusion and top up transfusions after birth. CONCLUSIONS: This study emphasises importance of red cell antibody screening for all multigravida antenatal women at registration of pregnancy and additionally at 28 weeks or later in high-risk cases irrespective of RhD status.

Three-phase DNA-origami stepper mechanism based on multi-leg interactions.

Nanoscale stepper motors such as kinesin and dynein play a key role in numerous natural processes such as mitotic spindle formation during cell division or intracellular organelle transport. Their high efficacy in terms of operational speed and processivity has inspired the investigation of biomimetic technologies based on the use of programmable molecules. In particular, several designs of molecular walkers have been explored using DNA nanotechnology. Here, we study the actuation of a DNA-origami walker on a DNA-origami track based on three principles: 1) octapedal instead of bipedal walking for greater redundancy; 2) three pairs of orthogonal sequences, each of which fuels one repeatable stepping phase for cyclically driven motion with controlled directionality based on strain-based step selection; 3) designed size of only 3.5 nm per step on an origami track. All three principles are innovative in the sense that earlier demonstrations of steppers relied on a maximum of four legs on at least four orthogonal sequences to drive cyclic stepping, and took steps much larger than 3.4 nm in size. Using gel electrophoresis and negative-stain electron microscopy, we demonstrate cyclic actuation of DNA-origami structures through states defined by three sets of specific sequences of anchor points. However, this mechanism was not able to provide the intended control over directionality of movement. DNA-origami-based stepper motors will offer a future platform for investigating how increasing numbers of legs can be exploited to achieve robust stepping with relatively small step sizes.

D or anti-D!!! Unblocking the dilemma of blocking-D phenomenon using acid elution.

Maternal IgG antibodies directed against fetal red cells can cause hemolytic disease in fetus and newborn manifesting as anemia and jaundice. Sometimes, these antibodies are so strong that they encapsulate the antigens on neonatal red blood cells and result in erroneous laboratory findings when tested. A requisition for double volume exchange transfusion was received for a term,3.1 kg female baby with neonatal jaundice at day 2 of life, born to a multiparous woman. The neonate was typed as AB RhD negative and the mother as A Rh D negative. The maternal sample tested positive for Indirect antiglobulin testing showing presence of Anti-D with IgG titer of 128. The direct antiglobulin testing for baby was strongly (4 +) positive. The strong DAT result with negative RhD typing for the neonate indicated towards the Blocking-D phenomenon. We attempted to resolve the Blocked-D case using acid elution, which revealed the presence of D antigen on the eluted neonate's red cells. The report emphasizes the importance of appropriate blood typing for neonates to provide prompt adequate care as a team by the departments of Neonatology and Transfusion Medicine.

Irregular erythrocyte antibodies among antenatal women and their neonatal outcome at a tertiary care hospital in Northern India.

BACKGROUND: Red blood cell alloimmunisation during the pregnancy is a significant cause for neonatal mortality and morbidity. This study was planned to determine the prevalence and specificity of irregular erythrocyte antibodies in antenatal mothers and their neonatal outcome. METHODS: In this observational study, blood grouping and red cell antibody screening of mothers were performed at first visit and after 28 weeks of gestation and positive cases were identified and followed up monthly till delivery by repeating antibody titre and middle cerebral artery-peak systolic velocity. After delivery of alloimmunised mothers, cord blood haemoglobin, bilirubin and direct antiglobulin tests (DAT) were analysed and further outcome of neonate was recorded. RESULTS: Among 652 registered antenatal cases, 18 multigravida women were found to be alloimmunised, accounting to prevalence of 2.8%. Most common alloantibody identified was anti D (>70%) followed by anti-Lea, anti-C, anti-Leb, anti-E and anti-Jka. Only 47.7% Rh D negative women received anti-D prophylaxis during previous pregnancies or whenever indicated. DAT was positive in 56.2% of neonates. Among nine DAT positive neonates, two early neonatal deaths due to severe anaemia were observed following birth resuscitation. Four antenatal mothers required intrauterine transfusion in view of fetal anaemia while three neonates received double volume exchange transfusion and top up transfusions after birth. CONCLUSIONS: This study emphasises importance of red cell antibody screening for all multigravida antenatal women at registration of pregnancy and additionally at 28 weeks or later in high-risk cases irrespective of RhD status.

The giant resectable carcinoma of gall bladder-a case report.

BACKGROUND: Gall bladder cancer (GBC) is the fifth most common malignancy in the gastrointestinal system and the most common malignancy of the biliary system. GBC is a very aggressive malignancy having a 5 year survival rate of 19%. Giant Gall Bladder (GGB) is an uncommon condition that can result from cholelithiasis or chronic cholecystitis and rarely with malignancy. CASE REPORT: A 65 year old lady presented with vague abdominal pain for 12 years and right abdominal lump of size 20 × 8 cms was found on examination. CT scan showed a circumferentially irregularly thickened wall (2.5 cm) of gall bladder indicative of malignancy. Per-operatively a GB of size 24 × 9 cm was noted and patient underwent radical cholecystectomy. It was surprise to find such a giant malignant GB with preserved planes. Histopathology, it was well differentiated adenocarcinoma of gall bladder of Stage II (T2a N0 M0). DISCUSSION: It is known that mucocoele of GB can attain large size, however chronic cholecystitis will lead to a shrunken gall bladder rather than an enlarged one. A malignant GB of such size and resectable is rare without any lymph node involvement or liver infiltration. Few cases of giant benign gall bladder have been reported in literature, however this appears to be the largest resectable gall bladder carcinoma reported till date as per indexed literature. CONCLUSION: Giant GB is an uncommon finding. They are mostly benign, however malignant cases can occur. Radiological findings may suggest features of malignancy and define extent of disease. Prognosis depends on stage of disease and resectability, irrespective of size.

Routine indirect antiglobulin testing of blood donors-a further step toward blood safety: an experience from a tertiary care center in northern India.

CONCLUSIONS: This scientific article emphasizes the importance of a policy for antibody screening of all blood donors as a step to further improve blood safety. We also report the incidence of red blood cell (RBC) alloimmunization in healthy blood donors obtained using a cross-sectional prospective study from September 2017 to January 2019 in the Department of Transfusion Medicine of a tertiary care referral and teaching institute in northern India. The indirect antiglobulin test (IAT) for unexpected RBC antibodies was performed by the conventional tube test with pooled group O RBCs on all donor units irrespective of their D status. Samples with positive IATs were sent to the Immune Hematology Reference Laboratory for further immunohematolo-gy workup, maintaining predefined optimal storage and transport conditions. Of the 10,390 donors studied, 9959 were males and 431 were females. The incidence of unexpected antibodies (antibodies other than those of the ABO blood system) among the blood donors was found to be 0.18 percent (19 of 10,390 with 25 alloantibodies). Of the 19 alloimmunized donors, 16 (84.2%) were male (alloimmunization rate 0.16%, 16 of 9959) and 3 (15.8%) were female (alloimmunization rate 0.69%, 3 of 431) (p = 0.01; chi-square test). In our study, the most frequent alloantibodies identified were of the Lewis blood group system (17 of 25 [68%] in 14 of the 19 alloimmunized donors). The second most common allo-antibodies belonged to the Rh blood group system (4 of 25 [16%] in 3 of the 19 alloimmunized donors), followed by those of the MNS blood group system (3 of 25 [12%] in 2 of 19 alloimmunized donors). Anti-K was found in one donor (1 of 25 [4%]). Based on the results of the study, we recommend that a policy of routinely performing IATs on all donor units, irrespective of their D status, be adopted as an essential component of safe blood transfusion practices.

Alarmed or unalarmed!! Donor red cell lysis during plateletpheresis procedure.

Plateletpheresis procedures are considered safe for donors and very few adverse events are reported. Donor related adverse events include hypo-calcemia due to citrate toxicity, vasovagal reactions, transient post-procedural decline in hemoglobin, hematocrit and platelet count. Kit related adverse events include damage of the kit or rarely mechanical damage of the red cells during procedure. This damage can lead to failure of the rinse-back of the red cells to the donors causing higher red cell loss in such donors. Here, we are reporting a kit related adverse event that led to donor red cell lysis during plateletpheresis procedure.

Primary carcinoma of the cystic duct: a case report and review of classifications.

BACKGROUND: The incidence of extrahepatic bile duct malignancies is about 2-3.6% of all gastrointestinal malignancies. Primary carcinoma of cystic duct is a rare condition comprising a fraction of all extrahepatic bile duct malignancies with less than 70 cases reported worldwide. Majority of these cases were reported from East Asia. There is paucity in such case being reported from Indian subcontinent. We present a case of primary carcinoma of the cystic duct encountered during laparoscopic cholecystectomy. CASE PRESENTATION: A 65-year-old lady presented to us with symptomatic gall stone disease. Investigations revealed a distended gall bladder with multiple stones. Patient was taken up for laparoscopic cholecystectomy, during surgery a stony hard structure was found at cystic duct-common bile duct junction which was not amenable for clear dissection. Procedure was converted to open, and the patient underwent cholecystectomy with resection of common bile duct with Roux-en-Y hepaticojejunostomy and regional lymphadenectomy. Histopathological findings revealed it to be moderately differentiated adenocarcinoma of the cystic duct. CONCLUSION: Primary carcinoma of cystic duct is a rare condition where early diagnosis can be difficult and if accidentally detected may add to surgeon's dilemma. Proper surgery with en-bloc resection of gallbladder, cystic duct, common bile duct, and regional lymphadenectomy is the mainstay of treatment. The prognosis of carcinoma of cystic duct is better than extrahepatic bile duct malignancies. The old classification system has outlived its time and is more rigid in definition which is not practical in advanced cases; the new classification systems of this century offer better insight into understanding the tumor characteristics and prognosis.

An Evaluation of Random Donor Platelets Prepared from Overnight Stored Buffy Coat Method: A Better Alternative to the 6-h Restriction.

Preparing maximum platelets from all daily blood donations is essential to maintain adequate stock of platelets that may be feasible by storing buffy coat overnight. The overnight stored buffy coat method is prevalent in Europe and was not studied much in India. Therefore, a comparative study was planned to compare RDPs prepared from fresh and overnight stored buffy coat methods. In this study total of eighty RDPs were prepared. Forty RDPs were prepared by each method by matching similar platelet counts in whole blood in pairs and avoiding other bias. Quality analysis was done in various hematological, biochemical, metabolic, and activation parameters and with National guideline. RDPs prepared by the overnight stored buffy coat method had statistically higher mean platelet counts, lesser WBC contamination, lesser glucose concentration, higher lactate concentration, lesser pH, and higher MPV & PDW than RDPs prepared by the fresh buffy coat method. P selectin levels were lesser in the overnight stored category. The platelet yield in RDPs (extraction from the whole blood) was more in the overnight stored category. Compared with the national guideline, 87.5% of units passed the quality criteria in the fresh category compared to 100% of units in the overnight category. RDPs prepared from overnight stored buffy coats not only had better quality parameters but also were in a less activated state. Because of better quality and feasibility in blood centers, the overnight stored buffy coat method may also be used along with the traditional preparation method.

Utility of COVID-19 Seropositive Plasma as Convalescent Plasma: An Immune and Neutralization Antibody Seroprevalence Analysis in Blood Donors for Future Potential Pandemic Readiness.

OBJECTIVES: To analyze the seroprevalence of SARS-CoV-2 IgG antibodies and neutralizing antibodies in blood donors during the second wave of the pandemic and to explore the utility of COVID-19 seropositive plasma as convalescent plasma. MATERIALS AND METHODS: In this study, 696 blood donors were tested for anti-SARS-CoV-2 IgG antibodies using a chemiluminescence assay. By blinding, 271 samples were chosen randomly for testing of neutralizing antibodies by enzyme-linked immunosorbent assay (ELISA) in duplicate among the 696 blood donors tested for anti-SARS-CoV-2 IgG antibodies, irrespective of the positivity or negativity of the result of the anti-SARS-CoV-2 IgG antibodies by chemiluminescence assay. IgG antibody levels were analyzed in signal-to-cutoff (S/Co), while neutralizing antibody levels were analyzed in percentage inhibition. RESULTS: The seroprevalence of IgG antibodies based on the S/Co for the positive results ≥ 1.00 was 82.75%, while the seroprevalence of neutralizing antibodies based on the percentage inhibition for the positive results ≥ 30% was 89.59%. Frontline workers (FLWs) and Covishield-vaccinated individuals showed higher levels of the anti-SARS-CoV-2 IgG antibodies regarding higher S/Co. In comparison, levels of neutralization antibodies regarding percentage inhibition were higher only in FLWs. Covishield-vaccinated donors elicited a statistically higher seroprevalence of anti-SARS-CoV-2 IgG antibodies compared to the Covaxin-vaccinated, while the seroprevalence of neutralizing antibodies was not statistically different among this group. There was a positive correlation (0.762) between anti-SARS-CoV-2 IgG antibodies and neutralizing antibodies, and almost all donors' of S/Co ≥ 9.5 had neutralizing antibodies. CONCLUSION: This study showed higher seroprevalence in the blood donor population compared to published seroprevalence in India's second wave of the pandemic. In the current study, 328 donors (47.12%) of the 696 screened donors were neither vaccinated nor had previous SARS-CoV-2 infection, but many had antibodies. The seroprevalence of neutralizing antibodies (96.42%) was higher than the seroprevalence of the anti-SARS-CoV-2 IgG antibodies (85.71%) in the donors who had previous infection of COVID-19. On the other hand, vaccinated donors showed similar immune responses for neutralizing antibodies and the anti-SARS-CoV-2 IgG antibodies. Higher IgG immune reactivity in S/Co showed a good correlation with neutralizing antibodies and can be used to screen whole blood donors for convalescent plasma donations.

Therapeutic Leukocytapheresis to relieve priapism due to leukostasis in CML patients: Our experience in a tertiary care centre.

Hyperleukocytosis in leukemic patients may cause tumour lysis syndrome, disseminated intravascular coagulopathy, and leukostasis, resulting in decreased tissue perfusion and increasing the risk of mortality. Since the myeloid blasts are larger than lymphoid blasts and are less deformable, complications of leukostasis are seen more frequently in myeloid leukemia. Priapism is a less common complication associated with leukostasis in leukaemia patients that should be treated as soon as possible to avoid ischemic injuries. Although chemotherapeutic drugs such as hydroxyurea and imatinib are used to treat hyperleukocytosis in CML patients, leukocytapheresis (LCP) can achieve rapid cytoreduction. Prophylactic LCP could not offer any advantage over aggressive chemotherapy, but therapeutic leukocyte depletion has a proven role in patients having symptomatic leukostasis due to high tumour burden. Three patients with ischaemic priapism were reported at our institute's emergency department, where detumescence could not be achieved by distal shunting or aspiration with phenylephrine instillation. The procedure of therapeutic LCP was performed in all three patients on an emergency basis, which resolved painful priapism by rapid cytoreduction.

Socio-Cultural Discourses of Children Engaged in Child Labor in India: A Macro-Ethnographic Study.

Background: Child labor is considered one of the main social problems that affect the community and has a physical and psychosocial impact on a child's health, growth, and development. The study aimed to describe the sociocultural discourses of children involved in child labor. A macroethnographic approach was used to collect the data from the selected community areas of Punjab, India. Materials and Methods: A community-based qualitative study using purposive sampling was carried out among children engaged in child labor (n = 8). The data were gathered through in-depth or semi-structured interviews and non-participant observation under four phases: community context assessment, egocentric network analysis, validation, and dissemination of study findings. The guide for consolidated criteria for reporting qualitative research (COREQ) was followed. Ongoing analysis was conducted using Spradley's analysis approach to identify the themes. Results: Thematic analysis resulted in the emergence of various themes related to child labor, namely, adaptability, money-centric attitude, sense of being underserved, social deprivation, work burden, reduced emotional expression, workaholism, abuse, family shoulder, family cohesion, and sense of industry. Apart from that, inhuman living and working conditions, as well as school deprivation among study subjects, were observed. Poverty emerged as the single most compelling factor for child labor in India. Conclusion: The study concluded that child labor had a negative impact on children's overall development. An effective intervention to stop child labor is if vulnerable children are identified through primary healthcare, and a relationship of trust is built that allows for the provision of health care, education, support, and referral to additional services outside the health sector.

Web-based E-survey in identifying current physiotherapy practices in paediatric burns.

INTRODUCTION: Physiotherapy practices in pediatric burns involves relaxation techniques and exercise training for attaining physical fitness, improving quality of life and, thereby to prevent primary and secondary complications. Physical activity and exercise act as a meaningful and purposeful tool for attaining physical fitness after burn injury. AIM: The aim of this e-survey is to identify the current practices of physiotherapy interventions in pediatric burn rehabilitation among physiotherapy students and professionals. METHODS: The e-survey was created with validated questionnaire using a Google Forms to gather the information related to current physiotherapy practices in pediatric burns. After the sample size estimation, survey link were sent to identified 144 physiotherapy students and professionals using social networking sites which includes, WhatsApp, Facebook and Instagram for this cross-sectional e-survey. The frequency and percentage of survey responses were analyzed. RESULTS: Among 144 identified participants, 62 participants completed the survey, resulting in the response rate of (43.0%). The findings of this study revealed that> 50% of physiotherapy students, and professionals, are performing pain and scar assessment, along with their regular physiotherapy management in children with burns. In addition to these, they also practice physiotherapy for burn conditions in pediatric intensive care units (PICU), pediatric wards, and clinical outpatient department (OPD) settings. CONCLUSION: Physiotherapy students and professionals have sufficient knowledge regarding pediatric burns complications, and also, they are well aware regarding recent physiotherapy practices in pediatric burn care management.

Development of a structured validated module to inculcate research skills in medical undergraduates.

Background Evidence-based research aids in decision-making in the health sector for developing health policies for prevention, diagnosis and treatment of diseases. Medical research is not taught in the undergraduate curriculum. Studies show that attributes of research knowledge, awareness and practical involvement in research are low among undergraduate students. We developed and validated a module and trained undergraduate students in research skills through an inter-ventional workshop using the structured module. Methods We did this participatory action research with a mixed-methods approach in the Department of Community Medicine at Adesh Institute of Medical Sciences and Research, Bathinda, Punjab. A structured module was developed by the core committee and validated internally and externally. Pilot testing of the module was done by delivering it in the form of a workshop to 46 students. For statistical analysis, percentage agreements, validity indices, median (interquartile range), satisfaction percentages and Wilcoxon sign test were used. Results The structured and validated module was established to have high face validity (>90%) and content validity (CVI=0.975). The module was successfully pilot tested for delivery through both onsite and online modes. The satisfaction percentage with the workshop was 91% and 100% and overall rating of the module was 74% and 91% by interns and MBBS students, and 100% by faculty. The scores of knowledge and skills were found to be significantly higher on all variables post workshop with p<0.001. All students scored satisfactory grades for research skills. Conclusions Teaching research using a structured validated module improved the knowledge and skills related to research among students. Both students and faculty were satisfied with the use of the structured module.