Deletion Screening and Point Mutation Analysis in Regions of the Duchenne/Becker Muscular Dystrophy Gene

Duchenne muscular dystrophy (DMD) is the commonest dystrophy with a birth incidence of one in 3000-3500 males, and approximately one third of all cases result from a new mutation. Affected males present with progressive muscle weakness and die as a result of respiratory or cardiac involvement in their late teens or early twenties. Becker muscular dystrophy (BMD) is a mild variety of the disease, and the most useful criterion for differentiating the two types is the age when patients become wheelchair bound. The gene is located on Xp21, and consists of more than 65 exons encoding a protein 3685 amino acids long, named dystrophin. About 65% of affected boys have a small submicroscopic gene deletion, heterogeneous in both the specific region and number of exons missing, that can be detected by Southern blot analysis using cDNA probes, and/or by sequence amplification with the polymerase chain reaction. Partial gene duplication accounts for the mutation in about 6% of the patients, while in the remainder point mutations are suspected An aim of this project was to screen DMD/BMD patients for deletions in exons 30 to 47 of the dystrophin gene covered by cDNA probe 5b-7, on Bglll digested DNA, and at the 3' end of the gene using the restriction enzymes Hind III and Bgl II in combination with cDNA probes 9, 10 and 11-14. Seventeen DMD/BMD patients were studied with cDNA 5b-7 and one deletion was detected having the distal end at exon 44 while the proximal end was in exon 20, in the region of cDNA probe 4-5a. Five cDNA deletions were detected having both end-points in the distal part of (lie dystrophin gene in a panel of thirty-six DMD/BMD affected males showing no deletion or duplication with the rest of the cDNA probes. Also, two cases of deletions which started in the region of cDNA 8 were identified to extend in the region of cDNA 9, with one of the two having the 3' end into the region of cDNA 10. No deletions were found with cDNA 11-14. Taking advantage of the different size of deletions detected, some of the Bgl II genomic fragments were related to Hind III genomic fragments of known orientation, and the order of the Hind III fragments in the region of cDNA 10 was rearranged. Also, the correlation between deletion and phenotype was examined and found to fit the reading-frame model proposed to explain the clinical difference in severity between DMD and BMD patients. For diagnostic purposes the detection of the molecular pathology of the disease can confirm the diagnosis of DMD/BMD in sporadic cases and offer direct accurate prenatal diagnosis in the family without the necessity for linkage analysis that requires DNA samples from key relatives. As Southern analysis takes five to ten days for results to be obtained assessment of the value and reliability of the polymerase chain reaction in performing deletion detection screening at multiple sites simultaneously was the next aim of the present study. Simultaneous amplification by PCR of exons 4, 8, 12, 17, 19, 44, 45, 48 and 51 of the dystrophin gene in 118 DMD/BMD DNA samples identified a mutation in 48. 3% of the patients and detected 86% of the deletions previously revealed by Southern analysis. Discrepancies were not found between the results obtained by the two methods of analysis. On the contrary, the problem of weak hybridisation was circumvented in two cases allowing the accurate mapping of the deletion end-points. Postnatal-detection screening was also performed in two patients and both results were confirmed by Southern analysis. The ability of PCR to amplify DNA fragments from small starting amounts of not necessarily good quality DNA permitted the amplification of DNA extracted from haematoxylin and eosin stained tissue sections, as well as the detection of a deletion in an individual whose DNA failed to produce detectable signal by Southern analysis because of degradation. Speed, sensitivity, specificity and efficiency characterise the method of multiplex amplification with the polymerase chain reaction, and make it ideal in the analysis of mutations in routine clinical practice as an initial screen to detect the molecular pathology of the disease. The final aim of this project was to scan regions of the DMD/BMD gene in affected males whose molecular pathology was still unknown, for new polymorphic sites or mutations that may account for the development of the disease and to compare and assess the different methodologies for mutational screening. (Abstract shortened by ProQuest.)

Pattern of referral and management of oro-dental problems in patients with cancer: a retrospective study

Background: General dentistry care is of paramount importance in maintaining oral hygiene of cancer patients. Methods: The data of patients registered at the dental wing in a tertiary care cancer center for one year period (2013) was used for the present analysis. All the cases were retrospectively analyzed for the pattern of referral, diagnosis of different oro-dental conditions, and different interventions done for the various oro-dental conditions.Results: A total of 798 patients were referred from various oncology wings for management of oro-dental problems, 50.1% patients were referred from head and neck oncology, 31% were from radiation department, 6.2% patients were from medical oncology and palliative care department each, 4% patients were from gynecologic oncology, and 2.5% patients were from screening wing. Acute gingivitis (62%) in pre treatment and mucositis (28%) in post treatment were major symptoms and signs for referral. 47% patients were managed with tooth extraction, grinding was done on 23%, scaling were done on 12.5%, desensitization was done on 8.7% patients, filling was done on 2.5% patients, prosthesis fitting in 2.5% patients, root canal treatment in 2.2% patients, and post surgical obturator fittings in 0.75% patients. Conclusion: Majority of cancer patients were referred for oro-dental care from head and neck oncology department with head and neck cancers and extraction of tooth pre radiotherapy and chemotherapy remains an important part of oro-dental care in cancer patients.

Relationship between serum ferritin and endocrinopathies in thalassemic children: A hospital-based study

Background: Thalassemia syndromes are a heterogeneous group of Mendelian disorders characterized by lack of/decreased synthesis of either α/β globin chains of hemoglobin. It results in ineffective erythropoiesis and lysis of mature red blood cells in the spleen. Thalassemic patients require multiple blood transfusions which led to iron overload. Objective: This study evaluated endocrinopathies in thalassemic patients of age 5–18 years and relationship with serum ferritin level. Materials and Methods: This is a hospital-based cross-sectional study on 70 transfusion dependent thalassemic children of 5–18 years attending the department of pediatrics, of a tertiary care hospital in Assam. Their height, sexual maturity rating, serum ferritin, calcium, alkaline phosphatase, phosphate, thyroid stimulating hormone, random blood sugar, and fasting blood sugar were recorded. The data were analyzed statistically and p <0.05 was considered to be statistically significant. Results: The study population (70) consisted of 40 males and 30 females. In this study, 91.67% (22 of 24 cases) had delayed puberty, 50% (35 of 70) were stunted, 37.14% (26 of 70 cases) were hypocalcemic, 31.43% (22 of 70) had hypothyroidism, and 2.85% (2 of 70) were diabetic. The mean serum ferritin level was 2845±859.40 ng/ml in delayed puberty while it was 2027.50±21.92 ng/ml (p>0.05) in thalassemic children with normal puberty. The mean serum ferritin in stunted thalassemic children was 2909.86±820.93 ng/ml and it was 1421.43±283.41 ng/ml (p<0.001) in normal height thalassemics. The mean serum ferritin in hypothyroid thalassemics is 2806.59±929.57ng/ml and is 1885.75±846.89 ng/ml (p<0.001) in euthyroid cases. The mean serum ferritin in diabetic thalassemics was 2772.50±1092.48 ng/ml and it was 2147.16±965.63 ng/ml (p>0.05) in nondiabetic thalassemics. Conclusion: Growth retardation and endocrinal complications significantly occur in transfusion-dependent thalassemic children. Adequate chelation therapy helps in controlling serum ferritin levels thereby enabling chronically transfused thalassemic children to grow normally without early endocrinal complications

Softwares no agronegócio : uma análise com enfoque contábil - financeiro

Trabalho de Conclusão de Curso (graduação)—Universidade de Brasília, Faculdade de Economia, Administração, Contabilidade e Gestão de Políticas Públicas, Departamento de Ciências Contábeis e Atuariais, 2018.O objetivo do estudo é analisar a eficácia dos softwares de gestão de propriedades rurais na produção de informações contábil e financeira. O estudo foi realizado através de uma observação analítica de 5 softwares atualmente comercializados, em que as características analisadas foram: forma de entrada de dados, capacidade de processamento, informações geradas, forma de apresentação, instrumentos de acesso e o grau de dificuldade de manuseio. A análise dos dados identificou que, os softwares são deficientes como sistemas de informações contábeis por não fornecerem informações contábeis e financeiras suficientes ao usuário, porém são eficazes no cumprimento de seus objetivos que é a gestão de fazendas

Knowledge of yoga among cancer patients and their attendants: a survey in a tertiary cancer centre in North East India

Yoga is an ancient Indian science that has evolved over thousands of years. Yoga deals with the physical, mental, moral and spiritual well-being of an individual.1 We have conducted an electronic survey amongst the cancer patients and attendants of our hospital about their knowledge of yoga. It’s a survey of 62 participants from 15/06/2021 to 18/06/2021 (before international yoga day on 21/06/21), 46.8% of the participants are male and 53.2% are female in the survey. The age of the participants ranges from 9 to 69 years. We have designed a set of ten questions, first six questions are designed to know their knowledge about the beneficial effects of the yoga, next two questions are about the misconceptions, cautions and guidance needs while performing yoga and the last two questions about the possibility of the practical application of it in their life. In our survey, we find that 93.5% of the participants think yoga reduces stress, 96.8% think yoga reduces fatigue, 95.2% opine yoga improves sleep, 93.5% feel yoga improves posture-related dysfunctions in cancer patients. Regarding chances of recurrence in cancer patients, 75.8% of patients think it has a negative impact on recurrence, 40.3% of participants have an opinion that yoga can cure cancer (no study till now proves that yoga cures cancer), and 90.3% feels that unguided yoga can cause injuries. At last, 93.5% patients feel yoga improve quality of life in cancer patients and 93.5% want to implement yoga in their daily life. From our survey, it seems participants have a fair idea of the benefits of yoga in cancer

Ultrasound evaluation of biopsy proven malignant breast mass

Background: Breast cancer is the most common malignancy in female worldwide. Ultrasound (US) is a safe and easily available modality for evaluation of breasts in females of all age groups. It can detect characteristic features of breast malignancy with high degree of accuracy. This study was undertaken with the aims to evaluate the ultrasonographic features of malignant breast mass, to identify the most commonly encountered gray scale ultrasound findings and to study the role of Doppler ultrasound in those cases.Methods: The study was conducted in Dr B. Borooah Cancer Institute, Guwahati from January 2018 to January 2019. Gray scale ultrasound including Doppler study was done in all the female patients coming with palpable breast lump or with nipple discharge and images were archived. The archived images of a total number of 108 patients with biopsy report positive for malignancy were retrospectively studied and the ultrasound findings were evaluated.Results: Most common gray scale ultrasound features for malignant breast masses were hypoechoic mass, taller than wide, irregular shape, having spiculated margins, neither posterior acoustic enhancement nor shadowing, presence of intralesional microcalcifications and surrounding echogenic halo. Hypervascularity, noticeable difference in waveform pattern between central and peripheral vessels, high resistance flow pattern with absent or reversal of diastolic flow were the common Doppler findings.Conclusions: Combination of Doppler with gray scale ultrasound has emerged as a very important technique in diagnosing malignant breast mass with high accuracy

Phosphorus recovery as struvite: recent concerns for use of seed, alternative Mg source, nitrogen conservation and fertilizer potential

Finite availability of phosphorus (P) resources makes recovery of this non-substitutable plant nutrient from alternative waste sources an increasingly attractive option of renewed interest. In this context, feasibility of struvite (MgNH4PO4·6H2O) recovery, an alternative P fertilizer is already demonstrated at laboratory scale from range of waste streams of farm, municipal and industrial origin, with reasonably high orthophosphate recovery efficiency (∼90%). However, apart from a few commercial extraction units using municipal sludge and urine, large scale struvite recovery is not widely adopted for many of these sources. Moreover, need of some research interventions that are restricting its profitable recovery are also highlighted by earlier studies. To increase recovery efficiency from identified potential sources in terms of cost and energy input, research focuses on some new aspects of the process such as prospects of alternative recyclable magnesium sources, different seed materials and their related issues, which are analyzed in this review. Prospects of nitrogen conservation through struvite recovery and fertilizer value of struvite considering its properties, comparative performance with conventional fertilizer and interaction with soil and plant growth are also critically reviewed

Oral premedication for pediatric anaesthesia: a comparison between midazolam and clonidine

Background:In children any anticipated surgery is associated with profound psychological stress to the patients. To allay this stress and anxiety, wide variety of pharmacological interventions are used.Methods: A prospective observational study was carried out between July’2005 and June’2006 in a tertiary care hospital in Assam. The study included children undergoing surgery under general anaesthesia in age group 2-10 years. Sixty children were divided into two groups, in one group patients received oral midazolam and other group received oral clonidine as pre-medicant.Results: In the midazolam group 23 (76.67%) children had no apprehension after parental separation and at induction (P˂0.001), sedation score at induction was significant higher in midazolam group versus clonidine group (93.33% versus 66.6%, P<0.01), clonidine resulted in a more stable pulse and blood pressure peri-operatively (P<0.05).Conclusion: Oral midazolam has better efficacy in terms of preoperative sedation and oral clonidine had stable hemodynamic profile and better analgesia in the clonidine group

Evaluation of bio-asphalt binders modified with biochar: a pyrolysis by-product of Mesua ferrea seed cover waste

With growing global concerns related to energy security and sustainability, interest in bio-fuels has increased significantly. Production of biofuel mostly begins with pyrolysis, a process that converts the biomass to liquid biooil, solid biochar and gases. A large amount of carbonaceous biochar is generated as a by-product during the pyrolysis. However, no major effort has been made in the past decade to utilize the biochar in pavement applications, especially as a bio-modifier to asphalt binders. In this study, an attempt was made to evaluate the carbonaceous biochar, obtained during bio-fuel production through pyrolysis of Mesua ferrea seed cover waste, as an asphalt modifier/extender to obtain bio-asphalts. Physical and chemical characterizations of biochar were carried out using scanning electron microscopy, Fourier transform infrared spectroscopy and energy dispersive X-ray analysis. This was followed by rheological characterization of asphalt binders (from two sources) modified with various concentrations of biochar (0, 5, 10, 15, and 20% by weight of binder). Flow behaviour, permanent deformation, and fatigue characteristics of bio-asphalts were evaluated and compared with control binders (no biochar). Aging susceptibility of biochar modified binders was also evaluated and compared for biochar contents and binder sources. Permanent deformation behaviour evaluated through multiple stress creep and recovery (MSCR) test indicated that the use of biochar improved the rutting resistance of binders. Improved binder performance against the effects of aging and deformation with the use of biochar makes it a promising asphalt modifier, especially for the warm tropical climatic conditions prevalent in India

Single Versus Multi‑visit Endodontic Treatment of Teeth with Apical Periodontitis: An In vivo Study with 1‑year Evaluation

Background: Apical periodontitis (AP) is one of the most prevalent diseases of the teeth. Treatment of AP is based on the removal of the cause, i.e., bacteria from the root canals. Achievement of adequate bacterial eradication in one appointment treatment remains a controversy.Aim: This prospective study was conducted with the objective to compare the periapical healing of teeth with AP treated in (a) single visit versus (b) two visits, either with or without Vitapex as an intracanal medicament.Subjects and Methods: Patients were selected randomly from the Department of Conservative Dentistry and Endodontics. Forty‑three patients (81 teeth) met the inclusion criteria, i.e., AP (both symptomatic and asymptomatic) visible radiographically size ≥2 mm × 2 mm, not suffering from any immune‑compromising disease, age between 16 and 65 years and tooth not accessed previously. Patients were randomly divided into three groups, i.e., single‑visit group (Group 1), multi‑visit group without any intracanal medicament (Group 2), and multi‑visit group with Vitapex as interim intracanal medicament (Group 3). Comparison was done radiographically using periapical index (PAI). The primary outcome measure was the change in periapical radiolucency after 1 year assessed by PAI scores. The Mann–Whitney U‑test was used to evaluate differences between groups at baseline (immediate postoperative) and at the 12‑month follow‑up evaluation. Change in PAI score for each group from baseline to 12‑month follow‑up evaluation was tested with the Wilcoxon signed rank test. The secondary outcome measures, proportion of teeth in each group that could be considered improved (decreased PAI score) or healed (PAI &lt;2), were assessed with the Chi‑square test.Results: No statistically significant difference in periapical healing was found between three groups.Conclusion: After 1‑year evaluation, no difference in periapical healing was found between single‑visit treatment and multi‑visit treatment groups with the given sample size.Keywords: 1‑year evaluation, Apical periodontitis, Multi‑visit treatment, Single visit, Vitape