Randomised controlled trial: role of virtual interactive 3-dimensional models in anatomical and medical education.

Purpose: Virtual interactive 3-dimensional models (VI3DM) and immersive virtual reality are implemented in medical education and surgical training. VI3DM allow learners to view and interact with a virtual 3D object and help in conceptualising learning objectives that demand high cognitive and visuo-spatial skills. However, the effects of VI3DM in medical education are unknown. We aimed to determine whether VI3DM are helpful in conceptualising complex anatomical structures. Materials and methods: We included 5 specimens, which were assessed by 200 first-year medical students categorised into experimental (n = 100) and control (n = 100) groups using a systemic randomisation method after matching for age and sex. The experimental group was given VI3DM as interventional learning resources while the control group was given 2-dimensional photographs as conventional learning resources for self-directed learning for 30 minutes. Participants completed a questionnaire before and after the learning session to assess their knowledge related to external features, attachments, and relations of anatomical specimens. Results: The scores of the experimental group improved significantly in the post-test compared to those of the control group for all 5 specimens included in the study (p < 0.05, confidence interval = 95%, unpaired student's t-test). Conclusions: VI3DM can help conceptualise external features, attachments, and relations of anatomical structures.

VI3DM allow interaction with and manipulation of 3D images of anatomical specimens on digital screens, web applications, mobile applications, and head mounted devices.Conceptualisation of anatomical structures requires processing of high cognitive and visuo-spatial information which can be eased by VI3DM.The anatomy can be visualised in high quality using digital 3D technology, which results in good perception of the third dimension and effectively increases the post-test score compared to the conventional method of learning, with higher engagement and satisfaction of the learners.Digital 3D models improve learners' engagement and compliance, specifically when the presented digital 3D models are interactive.

Legal Frameworks Upholding Deceased Individuals' Rights and Enabling the Use of Cadavers in Anatomy Education and Research: A Systematic Review.

The study of human cadavers is essential for teaching, advanced training, and research in medical and anatomical sciences. Medical institutions around the globe presently face a scarcity of cadaver supplies. For the majority of countries, unclaimed bodies are still the primary source of cadavers despite guidelines issued by the International Federation of Associations of Anatomists, which discourage the use of unclaimed bodies. This self-funded study aims to conduct a review of the existing national and international laws safeguarding the several rights of a deceased person. The study also reviewed the existing anatomy acts (and related acts) across various countries that facilitate cadaver supply for anatomy education and research. According to Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, an online search for publications in four medical databases (PubMed, Scopus, Web of Sciences, and Google Scholar) was conducted from 1950 till 2022. A research review protocol was registered in PROSPERO prospectively (registration ID: CRD42023469534) using the Mesh terms like cadaver, anatomy education, dead person and rights, body donation program, unclaimed bodies, and anatomy acts. After the application of the eligibility criteria, 149 publications were shortlisted. After reviewing all the shortlisted articles, laws, and guidelines, using the data extraction checklist prepared by both authors, five international laws, three supreme court case decisions, two high court case decisions, four Indian penal Code Sections, and 22 anatomy acts were selected. Articles from other medical databases could not be reviewed, which was the limitation of this study. The anatomy/human tissue/tissue transplantation acts and advisories from regulatory bodies of individual African, Middle-Eastern, and European countries could not be retrieved. The review findings will emphasize the distinctions between India's anatomy acts and those of other developed nations, thereby broadening our perspective as we propose a model anatomy act for uniform implementation across the country to streamline the procurement of cadavers.

Artery of Percheron, an Uncommon Variant of Posterior Cerebral Circulation: A Case Report.

The posterior communicating artery (PcomA), P1 and P2 segments of the posterior cerebral arteries (PCAs) give rise to numerous small branches that chiefly supply the thalamus and midbrain. Thalamic vascular supply is classically categorized into four regions: anterior, paramedian, infero-lateral and posterior. Despite significant variations and overlap in the blood supply, this traditional classification helps in understanding the vascular anatomy of the thalamus. Gerard Percheron extensively studied thalamic blood supply and described its anatomical variants depending on its origin. The artery of Percheron (AOP) is a rare anatomical variation of paramedian-mesencephalic arterial supply in which a solitary arterial trunk arises from the PCA and distributes bilaterally to both paramedian thalami and often to the rostral part of the midbrain. During routine dissection of the brain of a 46-year-old female in the department of anatomy, it was seen that thalamo-perforating artery (AOP) took origin as a single trunk from the P1 segment of the left PCA. The specimen was dissected and photographed for documentation and to see more details. The exact prevalence of AOP remains unknown, but various studies show that it can be present in 7% to 11.7% of subjects. Detailed knowledge of AOP anatomical variation is crucial for interpreting neuroimaging results or performing different neuro-endovascular techniques at the basilar bifurcation, particularly in patients with bilateral thalamic and midbrain infarctions.

Contribution of Anatomy in Forensic Age Evaluation: A Systematic Review.

The field of forensic anthropology is characterised by its ongoing development and growth. Forensic anatomy is a burgeoning discipline that focuses on the analysis and identification of both preserved and unpreserved human body parts, both in deceased individuals and the living. This subject plays a crucial role in establishing the four key factors of forensic anthropology, namely sex, age, race, and height. The objective of this research endeavour was to evaluate the significance of anatomical information in the process of forensic age estimation. The researchers established the inclusion criteria in accordance with the globally recognised Population, Intervention, Comparison, Outcome (PICOS) framework, as advised by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) recommendations. The research included many methodologies in order to ascertain the age. Upon conducting a comprehensive review of the existing literature pertaining to anatomical knowledge in the field of forensic age estimate, we have identified many notable applications. These include the utilisation of various anatomical features such as the dental pulp chamber, fingerprints, acetabulum, sternal end of the fourth rib, as well as hand and wrist bones for the purpose of age estimation. It is important for anatomists and other forensic scientists to engage in collaborative efforts to facilitate the exchange of ideas and ensure thorough investigations. This cooperation is particularly crucial in areas where anatomical sciences play a significant role in forensic science and investigation. Nevertheless, in order to mitigate the potential for estimating error, it is still advisable to use a multi-factorial evaluation approach that involves examining many body areas.

Is antero-lateral complex of knee joint of critical importance in restoring rotational instability in patients with anterior cruciate ligament tear?

Antero-lateral ligament complex (ALC) is a vital structure for maintaining rotational stability of the knee. Evaluation of ALC radiologically (MRI) is still unpopular in setting of anterior cruciate ligament injury. A dire necessity exists for the orthopedic surgeons in outdoor patient department settings to rule out involvement of ALC. So, that it can be addressed during operating for Anterior Cruciate Ligament injury. The authors have formulated an algorithm on a personal level and have implemented this screening program and initiated screening of young to middle aged patients reporting with rotational knee instability for ALC involvement before recommending final operative plan. This screening program which uses specifically devised physical tests have significantly reduced the number of underdiagnosed Antero Lateral Ligament tear.

The Impact of Virtual Interactive Three-Dimensional Model in the Conceptualization of the Anatomy of the Sacrum: A Randomized Controlled Trial.

Introduction Virtual interactive three-dimensional model (VI3DM) is an emerging technology with promising futures in medical education. It allows learners to view and interact with the three-dimensional (3D) object in an isolated virtual environment, as well as on screen-based platforms. This technology seems more helpful in understanding the learning objectives that demand high cognitive and visuospatial skills. The sacrum, part of the posterior wall of the bony pelvis, is a structure of interest to medical professionals and forensic experts. Understanding the anatomy and relations of the sacrum demands good spatial understanding. Hypothetically, virtual 3D models should help in learning the anatomy of the sacrum along with its relations and attachments. This study was conducted to find out the effect of low-cost digital 3D models on the anatomical knowledge of the study. Aims and objectives The goal of the work was to identify the role of virtual 3D models in the conceptualization of the anatomy of the sacrum. The study's objectives were to identify the impact of virtual 3D models on students' knowledge of the external features, relations, attachments, and joints formed by the sacrum. Material and methods Two hundred first-year medical students (168 males and 32 females) who participated in the study after providing informed consent were divided into two equal groups, a control group (n=100) and an experimental group (n=100), after matching the age, gender, and anatomical knowledge of the sacrum. We used two-dimensional (2D) images and virtual interactive 3D models of the sacrum as control and intervention, respectively, in this randomized controlled study. We conducted a post-test quiz after the 30-minute session of self-directed learning. Results The mean difference between the post-test score and the pre-test score of the experimental group (4.1±1.6 ) was significantly higher than the difference between the post-test and pre-test scores of the control group (2.5±1.2). The virtual interactive 3D model of the sacrum was significantly effective in the conceptualization of the sacrum anatomy. Conclusion A virtual interactive 3D model is an effective tool to conceptualize the anatomy of the sacrum and can be explored for its use in further complex anatomical structures. Digital 3D models can become a platform for the application of various virtual realities (VR) and artificial intelligences in medical education.

Real-world data on renal transplantations from a tertiary-care hospital in North India, in context of Indian regulatory act-transplantation of human organs and tissues act (THOTA): A retrospective analysis.

INTRODUCTION: Renal transplantation is the treatment of choice for patients suffering from end stage renal disease (ESRD). Indian regulations defined under Transplantation of Human Organs and Tissues Act (THOTA), 2014 restricts organ donations to near-related living donors to curb any malpractices like 'paid donors' in living-donor kidney transplantation (LDKT). The aim of our study was to look at real-world data of donor-recipient pairs and to identify relationship of donors (with their respective patients) and the common (or uncommon) DNA profiling methods used for supporting "claimed relationship" in accordance with the regulations. MATERIAL AND METHODS: The donors were categorized and grouped into near-related donor, donors other than near-related donors, swap donors and deceased donors. Claimed relationship was confirmed, commonly by HLA typing, using SSOP method. In few cases, which were uncommon (and infrequent), autosomal DNA analysis, mitochondrial DNA analysis and Y-STR DNA analysis were performed to support the claimed relationship. Data collected included age, gender, relationship, DNA profiling test method. RESULTS: Among the 514 donor-recipient pairs evaluated, numbers of female donors out-numbered male donors. The decreasing order of relationships in near-related donor group were wife>mother>father>sister>son>brother>husband> daughter>grandmother. 11.9% of donors were in the category of donors other than near-related donors. In 97.86% cases, the claimed relationship was supported by HLA typing and in just 2.1% cases autosomal DNA analysis>mitochondrial DNA analysis> Y-STR DNA analysis, in this order, were performed to establish relationship. CONCLUSION: This study brought out gender disparity with women out-numbering men as donors. Among recipients, access to renal transplant was largely restricted to men. As far as relationship of donors to recipients was concerned, mostly near-related family members, like wife, were donors and claimed relationship was almost always (99%) was corroborated by HLA typing.

Quadrangular Space Syndrome: a systematic review of surgical and medical therapeutic advances.

Background: The axillary nerve and posterior circumflex humeral artery are compressed in Quadrangular Space Syndrome (QSS), which can be treated with conservative approaches or surgical decompression in recalcitrant instances. There are no clear guidelines for determining which surgical method is optimal for treating QSS and other disorders that mirror QSS. Objective: The goal of this study is to grade and review past, current, and planned medicinal and surgical care modalities for QSS. Materials and Methods: The review protocol is registered with PROSPERO (ID: CRD42022332766). To identify recent advances in the methods/techniques of medical and surgical management of QSS, PubMed and Medline databases were searched until March 2022 for publications, including case studies, case reports, and review articles, using medical subject headings terms like quadrilateral space syndrome, surgical management, and medical management. Throughout the study, all the authors scrupulously followed a well-developed registered review process and the risk of bias in systematic reviews guidance tool. Data on proposed medical and surgical management methods/techniques were compiled, and each was analyzed based on the underlying neuro-vascular systems. Results: There were 88 items found in the first search. Following applying the inclusion and exclusion criteria, 16 papers were chosen for synthesis in the review study after a thorough assessment. Three studies (conservative and advanced) focused on medical care of QSS, while 12 articles (prior, current, and newer) focused on surgical management of QSS. Only four of the 15 studies reviewed proposed different surgical approaches/techniques for surgical decompression in QSS. Conclusions: There were two regularly used surgical procedures discovered, one anterior/delto-pectoral and the other posterior/ scapular. The anterior route is more technically straightforward and can be employed for surgical QSS decompression.

Persistent left superior vena cava: An anatomical variation.

The persistent left superior vena cava (LSVC) is a common anomaly of congenital heart disease. The presence of LSVC is commonly associated with other congenital cardiac anomalies such as atrial septal defect, tetralogy of fallot, aortic coarctation, ventricular septal defect and very rarely it occurs as an isolated finding. During a routine dissection for undergraduate students, a persistent LSVC along with variation in anterior cardiac vein and right septal pouch (RSP) was observed in heart of an approximately 48-year-old male cadaver. The persistent LSVC was draining into the right atrium via coronary sinus. The persistent LSVC is usually insignificant haemodynamically as commonly it drains into right atrium via coronary sinus, but incidental finding of LSVC is important to surgeons, interventional nephrologists and radiologists before placement of central venous access device. The insertion of central venous catheter via left internal jugular vein is difficult in presence of persistent LSVC. The right superior vena cava was normal. An anterior cardiac vein joined with the right marginal vein to form a common vein. The common vein opened into the right atrium. We also observed a RSP attached to the limbus fossa ovalis inferiorly which is a kangaroo pouch-like structure. A septal pouch is potential site predispose to thrombus formation and is more common on left side. In this case report we discuss embryology, clinical significance and review of literature related to persistent LSVC, anterior cardiac vein and SP.

Deltoid Intramuscular Injections: A Systematic Review of Underlying Neurovascular Structures to the Muscle and Proposing a Relatively Safer Site.

The deltoid is the preferred site for intramuscular injection (IMI) because of its easy accessibility for drug and vaccine administration. Government immunization advisories, standard anatomy textbooks, and researchers have proposed various injection techniques and sites, but specific guidelines are lacking for the administration of IMIs in the increasingly used deltoid site. This study analyzes the procedures of administering IMIs in the deltoid related to the neurovascular network underlying the muscle and proposes a preferred site with the least chance of injury. The review protocol was submitted with PROSPERO (ID: 319251). PubMed, Google Scholar, and Websites of National Public Health Agencies were searched from 1950 up to 2022 for articles, advisories, and National Immunization Guidelines using Medical Subject Headings (MeSH) terms, including IMIs, deltoid muscle, safe injection sites, to identify recommendations for safer sites and techniques of administering deltoid IMIs. All the authors strictly adhered to a well-developed registered review protocol throughout the study and followed the risk of bias in systematic reviews (ROBIS) guidance tool. The proposed sites and landmark data were tabulated, and each site was analyzed based on the underlying neurovascular structures. Data were depicted by self-generated images. The initial search identified 174 articles. After applying the inclusion and exclusion criteria, 57 articles were shortlisted. Out of the 39 selected articles, 18 focused on the administration of deltoid IMIs, whereas seven focused on the variations in the underlying neurovascular structures in proximity to the deltoid muscle. The remaining 14 articles were the immunization guides issued by the National Public Health Agencies of the Government of India and abroad, whose data was used for comparison. Twelve deltoid IMI sites and techniques were identified. A site 1-3 fingerbreadths/5 cm below the mid-acromion point (7 studies); mid-deltoid site/densest part of the deltoid (1 study); a site at the middle third of the deltoid muscle (1 study); triangular injection site (1 study). Limitations included the unavailability of free access to complete text in many articles resulting in exclusion. The area around the shoulder joint and up to the lower level of the intertubercular sulcus is highly vascularized by the presence of many anomalous arterial patterns. To avoid injury, a safer site is proposed of 5 fingerbreadths/10 cm below the midpoint of the lateral border of the acromion. The authors received no specific funding for this study except for the journal publication charges.

Revisiting the Surgical Approaches to Decompression in Quadrilateral Space Syndrome: A Cadaveric Study.

Background Quadrangular space syndrome involves compression of the axillary nerve and posterior circumflex humeral artery. In a few cases, its management requires surgical decompression. The current study reviews the surgical approaches used in the decompression of neurovascular structures and presents our reflections and recommendations. Methodology In this study, four human cadavers were used for dissection of the axillae and the scapular region by the senior residents of the Department of Anatomy and Department of Orthopedics. The residents dissected the quadrangular space in the eight upper limbs using anterior and posterior surgical approaches. Results To identify the quadrangular space and secure its contents, the posterior approach was recognized as the easier and quicker method by both Anatomy and Orthopedic residents; however, it may result in increased postoperative morbidity. Although the anterior (deltopectoral) approach involves more skill, it reduces postoperative morbidity. Conclusions The anterior (deltopectoral) approach with suggested modifications can be an effective method in the surgical decompression of quadrangular space syndrome. The authors suggest more cadaveric studies to provide anatomists and surgeons with the opportunity to practice and evaluate older and newer surgical approaches.

Diagnostic value of different interleukins and procalcitonin in critically ill patients admitted with suspected sepsis.

BACKGROUND: : Many biomarkers have now been studied such as C-reactive Protein (CRP), procalcitonin (PCT), etc., and are widely used for the diagnosis of sepsis in clinical practice which may determine the appropriate antibiotic treatment. A flowcytometric cytokine bead array (CBA) assay has now been used to determine multiple interleukins (IL), simultaneously. The aim of this study was to determine the cytokine (IL2, IL4, IL6, IL10, TNFα, INFγ, and IL17) profiles of interleukins in plasma of sepsis patients by using multiplex Flowcytometric CBA array assay. MATERIALS AND METHOD: s: A total of 99 consecutive patients admitted with the suspected sepsis were studied. PCT concentrations were measured by using the enzyme-linked fluorescent immunoassay (ELFA) technique and flow cytometry-based BD™ CBA Cytokine Kit was used to evaluate levels of 7 cytokines [IL-2, IL-4, IL-6, IL-10, Tumour Necrosis Factor (TNF), Interferon- γ (IFN-γ), and IL-17A]. RESULTS: Microbiologically defined infection (MDI) demonstrated a positive culture report in 79/99 (79.7%) of patients. The IL6 [1873.7 (4-5000)] and IL10 [(154.7 (0-1764)] levels were significantly higher in septic patients than those in the negative MDI IL6 [901 (4-5000)] and IL10 [110.4 (4-1372)] levels. The AUROC value of IL6 [0.66 (0.53-0.79)] was found to be the highest among all followed by IL10 [0.65 (0.51-0.79)], IFNγ [0.63 (0.51-0.77)], PCT [0.61 (0.48-0.75)], and TNFα [0.55 (0.42-0.69)]. CONCLUSION: Our study suggests that that IL6 is substantially more economical and can reduce the investigation cost to half as compared with the procalcitonin assay.

The utility of the Ion Torrent PGM next generation sequencing for analysis of the most commonly mutated genes among patients with colorectal cancer in India.

Background: The requirement for the mutation analysis for Kirsten rat sarcoma viral oncogene (KRAS) in colorectal cancer (CRC) is rapidly increasing as it is a predictive biomarker and also, its absence signifies response to anti-epidermal growth factor receptor (anti-EGFR) antibody treatment. The aim of our study was to investigate the pathological diagnosis and distribution of KRAS mutations in colorectal cancer with the use of next generation sequencing platform (Ion Torrent). Methods: A total of 56 CRC samples were tested to identify the genetic mutations, especially KRAS using the primers which included ~2800 COSMIC mutations of 50 oncogenes. Ion Torrent personal genome machine (semiconductor-based sequencing) was used for the sequencing and analysis. Along with KRAS, other 49 genes were also studied for COSMIC mutations. Results: KRAS mutation 25 (44.6%) had the highest frequency, followed by TP53 10 (17.9%) and PIK3CA mutation 4 (7.1%). Of all the KRAS mutations identified, mutations in codon 12 were most frequent followed by mutations in codon 13 and 61. The most frequent substitution was glycine to aspartate mutation in codon 12 (p.Gly12Asp) followed by glycine to valine (p.Gly12Val). Combinations of mutations were also studied. Our study revealed that seven cases (12.5%) had both KRAS and TP53 mutations (highest of all the combinations). Conclusion: The analysis of KRAS mutation frequency and its mutational subtype analysis in human CRCs by using semiconductor-based platform in routine clinical practices have been performed in Indian population. The findings were similar to earlier published reports from the Western literature.

Prospective multi-centric study to analyze pre-transplant compatibility algorithm for live-related donor kidney transplant in Indian setting: the "Delhi approach"!

BACKGROUND: Since no single test is always accurate and sensitive, two or more tests are used to increase the precision of evaluation. Different algorithms have been proposed by centers in Leiden, Basel, Vienna and Minnesota, etc. With an intention to develop an optimal algorithm for India, we evaluated pre-transplant compatibility tests for live-donor kidney transplants. Three tests complement dependent cyto-toxicity cross-match (CDCXM), flow-cytometry cross-match (FCXM) and anti-HLA antibody screening (HAS) were performed and confirmed by the anti-HLA antibody identification (HAI) assay in a multi-centric trial (three transplant centers) in India. MATERIALS AND METHODS: All prospective recipients (and their potential donors) underwent low-resolution HLA typing as well as CDCXM, FCXM and HAS assays. In addition, HAI {single antigen bead assay; (SAB)} was done for all recipients to identify possible anti-HLA antibodies. In a virtual cross-match (VXM), antibody specificity was mapped to donor HLA type to determine donor-specific antibodies (DSA). Only patients without DSA were cleared for the transplant. Alternatively, patients with DSA were offered an exchange in the kidney paired donation (KPD) program. The screening results (CDCXM, FCXM, and HAS) were analyzed, individually as well as in combination of screening assays (CDCXM+HAS, CDCXM+FCXM, and FCXM+HAS) and the results were compared with those from the HAI test. RESULTS: Out of 100 patients, 69 were males and 31 were females; 85 recipients (85%) underwent a kidney transplant. The sensitivity of CDCXM was only 12.1% and the specificity of CDCXM was 100%; whereas the sensitivity of FCXM was 84.8% and the specificity of FCXM was 89.6%. The sensitivity and specificity of class I HAS was 88.2% and 84.3%, respectively. The sensitivity and specificity class II HAS was 88.0% and 80.0%, respectively. However, when both class I/II HAS were tested together the sensitivity increased to 97.0% and the specificity to 82.1%. Similarly, the sensitivity of combined FCXM+HAS had the sensitivity of 100% and the specificity of 76.1%; CDCXM+FCXM had the sensitivity of 84.8% and the specificity of 89.6% and CDCXM+HAS assays reached 97% with the specificity of 82.1%. CONCLUSIONS: Our results showed that the algorithm of FCXM with HAS produced the best sensitivity of 100%. The specificity of 76.1% indicate that the combined FCXM+HAS assays may detect up to 24.9% false positive results. We suggest that these false-positives may be easily resolved by performing the virtual crossmatch based on HAI (SAB) results. In our reflex testing algorithmic approach only 49% patients needed HAI (SAB). Finally, our results suggested that the CDCXM assay may be discontinued in pre-transplant workup owing to its very low sensitivity (12.1%).

Prevalence of Clinically Significant anti-HLA Antibodies in Renal Transplant Patients: Single-center Report from North India.

INTRODUCTION: Solid organ transplantation is the preferred therapeutic modality of treatment in patients affected by terminal organ failures. Human leukocyte antigens (HLAs) plays an important role in graft survival. In many of the cases of rejection, antibodies are directed against HLA antigens expressed on the cells of the transplanted organ. Pre-transplant compatibility testing involves the use of different methodologies for the determination of anti-HLA antibodies. Luminex single-antigen bead (SAB) assay demonstrates higher sensitivity and specificity in detecting anti-HLA antibodies. The aim of this study was to determine the prevalence of anti-HLA antibodies in pre-transplant work up recipients, planned for renal transplant at a tertiary care center in India. METHODS: 1640 patients visiting tertiary care hospital for pre-transplant compatibility testing were screened with complement-dependent micro-lymphocytotoxicity crossmatch (CDC-XM) and flow cytometric crossmatch (FC-XM). The patients positive for either or both screening tests were assayed with the Luminex SAB tests in order to establish defined antigen specificity of the alloantibodies and determining donor-specific antibody (DSA). RESULTS: The two most frequent antibodies identified in each A, B, C locus of HLA class I were -A*24:03 (43.9%), A*25:01 (36.6%), B*57:01 (40.3%), B*15:12 (37.1%), C*17:01 (61.9%), C*07:01 (52.4%) and in DR, DQ DP locus in HLA class II were DRB1*09:01(40.0%), DRB1*14:04(37.6%), DQA1*04:01/DQB1*03:03 (58.4%), DQA1*05:01/DQB1*03:01 (55.1%), DPA1*02:01/DPB1*17:01 (55.0%), DPA1*02:01/DPB1*05:01 (45.0%). CONCLUSION: This study has found the prevalence and specificity of anti-HLA antibodies in north India.

Detection of T and B cells specific complement-fixing alloantibodies using flow cytometry: A diagnostic approach for a resource limited laboratory.

BACKGROUND AND OBJECTIVES: Various methods have been reported for the detection of antibodies in recipient sera, which can be human leukocyte antigens (HLAs) or non-HLA specific, complement- or noncomplement fixing, as well as donor T (HLA-Class-I) and/or B cell (HLA-Class-I and II) specific. These alloantibodies play a pivotal role in antibody-mediated renal transplantation rejection. Deposition of C4d in peritubular capillaries of a kidney biopsy is a marker of antibody-mediated rejection. The C4d flow-panel reactive antibodies (PRAs) are a screening method for HLA-specific and complement fixing antibodies. However, the method is limited by the lack of donor specificity. DESIGN AND SETTINGS: Here, we present a new and simple flow cytometric method referred to as C4d-flow cytometry crossmatch (C4d-FCXM) for the detection of donor-specific (T and/or B cell) and C4d-fixing alloantibodies. RESULTS: The method was applied in a series of clinical cases and judged to be useful. The method may limit unwanted deferral of the donor due to positivity in C4d Flow-PRA and/or FCXM and may be helpful in prediction of antibody mediated rejections. Furthermore, this method can provide information pretransplant in contrast to kidney biopsy and C4d evaluation done posttransplant. CONCLUSIONS: We postulate that this method incorporates most of the features of all the available modalities (i.e., National Institute of Health-complement dependent lymphocytotoxicity, FCXM, cytotoxic FCXM and C4d-flowPRA) yet cost-effective and best suited for resource-limited laboratory/ies which is a common scenario in developing countries.

Phenotypic and genotypic characterization of phosphate solubilizing bacteria and their efficiency on the growth of maize.

Phosphate solubilizing bacteria (PSB) has ability to convert insoluble form of phosphorous to an available form. Applications of PSB as inoculants increase the phosphorus uptake by plant in the agriculture field. In this study, isolation and identification of PSB were carried out in Indian agriculture field (Nainital region, Uttarakhand). A total of 8 phosphate solubilizing bacterial colonies were isolated on the Pikovskaya's (PKV) agar medium, containing insoluble tricalcium phosphate (TCP). The colonies showed clear halo zones around the bacterial growth were considered as phosphate solubilizers. Out of 8 bacterial isolates, 3 isolates showed high phosphate solubilization index (PSI) ranged from 4.88 ± 0.69 to 4.48 ± 0.30, lower pH ranging 3.08 ± 0.08 to 3.82 ± 0.12 and high phosphate solubilization varied from 305.49 ± 10 µg/ml to 277.72 ± 1.45 µg/ml, were selected for further characterization. Based on the 16 S rRNA gene sequence analysis A4 isolate and H6 isolate were closely related to Alcaligenes aquatilis (99%), and C1 isolate was closely related to Burkholderia cepacia (99%). In addition, pot examination also showed the greatest efficiency in promotion of maize growth compared to uninoculated plant. Isolated PSB were able to produce different organic acids (such as gluconic acids, formic acid, and citric acid) in the culture supernatant and may consider as the principle mechanism for phosphate solubilization. This study clearly indicates that A4, C1 and H6 isolates may use as a biofertilizers in ecological agricultural systems instead of synthetic chemicals and may help to sustain environmental health and soil productivity.