Loop-mediated isothermal amplification assay for the rapid detection of Anaplasma marginale in cattle based on major surface protein 5 gene

Background & objectives: Timely intervention is needed to minimize the economic losses of vector-borne bovine anaplasmosis which can be possible by the isothermal amplification assay. Methods: Anaplasma marginale in the cattle of south Gujarat, India was detected in the PCR and LAMP by amplifying the fragment of msp5 gene. The PCR product was digested with EcoRI, and sequenced to confirm its pathogen specific detection. Results: Species specific PCR observed a band of 457 bp of msp5 DNA following 1% agarose gel electrophoresis. Positive LAMP reaction turned into yellow colour while negative sample depicted original pink colour. A detection limit of PCR and LAMP was up to 10-6 and 10-8 of the original genomic DNA of A. marginale, respectively. A single cut site of EcoRI was observed in the PCR product. Current msp5 DNA sequences of A. marginale (MW538962 and MW538961) showed 100% homology with the published sequences. Monophyletic lineage type relationship was observed with high bootstrap proportion among the msp5 DNA sequences of A. marginale in the phylogram. Prevalence rate of A. marginale was significantly higher (p<0.05) in the PCR [43/280 (15.36%)] and LAMP [62/280 (22.14%)] than the microscopic technique [17/280 (6.07%)]. Diagnostic sensitivity, specificity, positive and negative predictive values at 95% CI for LAMP assay with respect to PCR were 93.02%, 90.72%, 64.52% and 98.62%, respectively. Interpretation & conclusion: Thus LAMP can be a practical alternative to the PCR for the diagnosis of A. marginale infection in the cattle even in field condition

A Sole t(10;19)(p11.2;p12) in AML-ETO negative AML-M2 paediatric patient: First novel case from India

Acute myeloid leukemia (AML) is a heterogeneous group of disorders with regards to its pathology and molecular genetics features. The translocation t(8;21)(q22;q22), which results in the fusion of the AML and ETO genes, is a recurrent aberration in AML, preferentially correlated with FAB- M2, and has the highest incidence in childhood AML. Because of the favorable prognosis, the evidence of the t(8;21) or the AML1-ETO fusion gene is mandatory in most of the therapy trials, allowing the stratification of the patients to the correct risk group in terms of treatment. Here, we describe a novel case of sole translocation t(10;19)(p11.2;p13) in a AML1-ETO negative AML-M2 patient. In general, this translocation is previously observed with combination of complex translocations, but sole abnormality was not previously observed. This is a novel translocation and not observed previously as a sole abnormality in any AML case. Hence, the functional role of this translocation is still unknown. Short term bone marrow culture was carried out for conventional cytogenetics and karyotype was 46, XX,t(10;19)(p11.2;p12) in all 20 metaphases analyzed. To confirm this translocation FISH with Whole chromosome paint probe was applied and results confirmed the translocation between chromosome 10 and 19. To the best of our knowledge, this is the first novel case of sole t(10;19) in a paediatric AML-M2 patient with AML-ETO negative fusion. Patient expired within a week. Therefore, the present case challenges the view that the occurrences of sole and new novel translocation require more such cases to be studied in large cohort which is generally an indication for poor prognosis.

Post Mortem Study Of Injuries to the Internal Neck Structures in Cases of Death Due To Hanging.

Virtually all hangings are suicidal in nature and all ligature strangulations are homicidal in nature. So for the purpose of police investigation differentiation between two is very important and necessary. It is generally said that deaths due to hanging are devoid of any injury to the internal neck structures while in strangulation these injuries are always present. So injuries to the internal neck structures are sometimes used as differentiating factor between hanging and strangulation along with other factors. Aim: The aim of this study is to know the prevalence of injuries to the internal neck structures in deaths due to hanging. Method: This study was conducted at mortuary of the civil hospital, Ahmadabad in the year 2013. Total 40 cases of deaths due to hanging were randomly selected. Result: Out of these 40 cases 26(65%) were male and 14(45%) were female. 22 (55%) cases were showing injuries to the internal neck structures in the form of haemorrhage in to the soft tissues and strap muscles or fracture of superior horn of thyroid cartilage or greater cornue of hyoid bone. However these injuries are mainly found present beneath the ligature mark except some indirect injuries e.g. haemorrhage at the origin of sternomastoid muscle and avulsion fracture of greater cornue of hyoid bone due to over stretching of thyro-hyoid membrane. These injuries in cases of hanging are not extensive as found in cases of death due to strangulation. In one case extension distraction fracture of cervical spine at c3-c4 level was found which very rare finding in suicidal hanging is. Conclusion: From the present study it is evident that injuries to the internal neck structures are not very uncommon in the hanging. Though the extent and frequency of injuries are less compare to the ligature strangulation.

Radiological Study of Epiphyseal Fusion Process at Lower End Of Femur In Age Group of 12-20 Years.

Introduction: Age estimation in living as well as dead is a prerequisite for personal identification and it is increasingly important in criminal and civil matters. The growth of the human skeleton is of major importance for the aging process as the appearance of ossification centers and union of epiphysis relate to a fairly definite sequence and time table that makes skeletal maturity a reliable age indicator according to sex and ethnical differences1,2,3,.This present work is carried out to study the epiphyseal fusion of lower end of femur bone in relation to age, sex, physical development, and nutritional status. Materials and Methods: The present study was conducted in the Department of Forensic medicine & Toxicology and Department of Radiology at B.J. Medical College & Civil Hospital, Ahmedabad during the year 2010-2012 on 160 subjects, 80 males and 80 femalesfrom the age-group of 12-20 years. Results: The epiphyseal fusion in both male and female at lower end of femur starts by the age of 13-14 years and complete by 17-18 years in male and 16-17 years in females.It is not found any effect of the dietary habit, height, and weight on the timing of epiphyseal fusion of lower end of femur. Conclusion: From our study we may conclude that the epiphyseal fusion in both male and female at lower end of femur starts by the same age and completes earlier in females than males.There is no effect of diet, height, and weight on epiphyseal fusion of lower end of femur.

Ankle Brachial Pressure Index: As a predictor of peripheral arterial disease in diabetic & non diabetic subjects.

Background: Peripheral arterial disease is known to be associated with the diabetes and ankle-brachial pressure index (ABPI) can be used to screen such patients having PAD. Aims & Objective: To highlight the application of ankle brachial pressure index as a routine procedure and to assess the feasibility of using ABPI in detecting PAD in the patients of diabetes mellitus. Material and Methods: 30 Diabetic and 30 Non diabetic patients were recruited. Details regarding anthropometric measurements, blood sugar levels, lipid profile, Edinburgh questionnaire, peripheral pulse examinations etc. were filled in preformed questionnaire. Ankle-brachial pressure index (ABPI) was measured with the help of Handheld Doppler machine. Descriptive statistics in the form of frequency and percentage is used for description of data. Chi-square test was used for comparison of events in two groups. Results: PAD was more common in old age but found not to be associated with gender. Body mass index, surface area and wait hip ration was significant more in PAD group as compared to non-PAD. All blood sugar parameters [Fasting blood sugar (FBS), postprandial blood sugar after 2 hours (PP2BS) and Glycosylated haemoglobin (HB1AC)] were significantly more in PAD group as compared to non-PAD. High density lipoprotein was significantly less in PAD group as compared to non-PAD. Triglyceride, Total Cholesterol and Low density lipoprotein was significantly more in PAD group as compared to non-PAD. ABPI was significantly less in diabetics as compared to non-diabetics. More diabetics were in having PAD as compared to non-diabetics. Conclusion: PAD is found to be associated with more weight, adverse lipid profile and diabetes. ABPI can be used to screen the PAD in diabetic patients.

Trisomy 8 in leukemia: A GCRI experience.

Trisomy of chromosome 8 is frequently reported in myeloid lineage disorders and also detected in lymphoid neoplasms as well as solid tumors suggesting its role in neoplastic progression in general. It is likely to be a disease-modulating secondary event with underlying cryptic aberrations as it has been frequently reported in addition to known abnormalities contributing to clinical heterogeneity and modifying prognosis. Here, we share our findings of trisomy 8 in leukemia patients referred for diagnostic and prognostic cytogenetic assessment. Total 60 cases of trisomy 8, as a sole anomaly or in addition to other chromosomal aberrations, were reported (January 2005–September 2008). Unstimulated bone marrow or blood samples were cultured, followed by GTG banding and karyotyping as per the ISCN 2005. Patients with +8 were chronic myeloid leukemia (CML) (36), acute myeloid leukemia (AML) (17), and acute lymphoblastic leukemia (ALL) (7). In 7 patients, trisomy 8 was the sole anomaly, whereas in 6 patients +8 was in addition to normal clone, in 47 patients, the +8 was in addition to t(9;22), t(15;17), and others, including 3 with tetrasomy 8. Only one patient showed constitutional +8. The present study will form the basis of further cumulative studies to correlate potential differential effects of various karyotypic anomalies on disease progression and survival following a therapeutic regime. To unravel the role of extra 8 chromosome, constitutional chromosomal analysis and uniparental disomy will be considered.

Pulmonary functions in different phases of menstrual cycle and its relation with serum progesterone levels.

Background & objectives: Cyclical changes in the hormonal levels determine different phases of menstrual cycle. These variations in hormonal levels are also responsible for variations in functional parameters of many systems in the body. Many studies have highlighted the changes in respiratory parameters in different phases of menstrual cycle. In this study, we have explored the factors that influence lung function in adolescent girls focusing on the hormonal factors related to the menstrual cycle. Methods: 36 adolescent girls with normal menstrual cycle consented for the study. Pulmonary functions and serum progesterone were measured during follicular and secretory phase of menstrual cycle after a detailed menstrual history and screening for medical illness. Results : show a significantly higher serum progesterone and FVC, FEV1 and PEFR during secretory phase. There was a strong positive correlation of Serum progesterone in secretory phase with FVC and negative correlation with FEV1%. Interpretation & conclusion: This improvement of pulmonary function during secretory phase is due to increase in serum progesterone levels which have a dual effect of overall smooth muscle relaxation and hyperventilation. Supplement of progesterone may be useful to avert premenstrual asthma and enhance the performance.

Original research paper radiological study of disappearance of tri-radiate cartilage in pelvic-bone in Gujarati population.

For medico-legal purposes evidence of age is very essential in various types of cases such as Identification, Consent, Rape, Attainment of Majority, and Eligibility for Employment, etc. Various work done on the study of age determination by the study of appearance of ossification centers and epiphyseal union of different bones in India as well as abroad indicate a reasonable variation. Majority of them conclude that, the ossification and epiphyseal union of a bone vary in different part of the world. To the best of our knowledge very little work has been done on the study of disappearance of tri-radiate cartilage out of various bony parts useful for age estimation. The present study comprising of 50 subjects ( male=35; female=15),was carried out during the period January 2000 to July 2001 at the Forensic Medicine department and Department of radiology, B.J. Medical College, Ahmedabad.

Epiphyseal fusion at lower end of radius and ulna valuable tool for age determination.

Age estimation is an important task and valuable tool to assist in many civil and criminal procedures, especially in developing countries like India where illiterate population is not aware of the importance of registration of births or the record of registration may not be properly maintained. Physical methods for age estimation are not accurate and eruption of teeth (except third molar) is complete by the age of 16 years. Epiphyseal fusion of long bones is relatively constant in timings and important for age estimation in such cases up to the age of 22 years. This present prospective study is carried out at Forensic Medicine Department of B. J. Medical College, Ahmedabad during the year 2009-10 on 104 subjects of either sex with known age from 15-21 years. It was noticed that the epiphyseal fusion at lower end of radius and ulna processes and progresses bilaterally symmetrical, begins at the age of 16-18 years and completes by the end of 20 year. It was further noticed union at lower end of ulna occurs in advance of radius and females show union in advance of male subjects.