267 research outputs found
Molecular analysis of the promoter and the core promoter region Of the smn2 gene in circumscribing the clinical severity of sma
Spinal muscular atrophy (SMA) is a neurodegenerative disorder caused by the absence of the full
length SMN protein (FL-SMN) as a result of mutation or deletion of SMN\ gene. The isoform to
this gene, SMN2, with mutation in 1 base pair, encodes for 10% of FL-SMN protein and is
reported to decrease the severity of the disease when there is an increase gene dosage. There are
3 clinical types of SMA; type I, type II and type III. Type I SMA is the most severe type and
only a small amount of FL-SMN protein is present in these individuals. In this study it was
hypothesized that the variation in the DNA sequence in the promoter region of the SMN2 gene
could be the possible cause of difference in clinical severity of SMA. To verify this hypothesis, a
total of ten (10) SMA patients from different clinical types were selected from the retrospective
pool data of the SMA patients and their DNA was extracted. The promoter regions of the SACV2
gene in these patients were screened for the presence of any variation, firstly within the proximal
promoter region (Pro) which also contains a reported CR.E-II element followed by the screening
of the entire promoter region. The screening of the entire promoter was performed in two steps.
Firstly, the regions within the promoter with computational cis acting elements were identified
(using Cister analysis) and then the DNA sequencing was performed using specific designed
primers. Secondly, the regions other than cis acting elements were screened for the presence of
any variations. The bioinformatics analysis also revealed the probability for crucial transcriptional factor bindings sites, novel TSS at -557 bp and a TATA box at -411 bp within the
15 ORFs and 24 nested ORFs. The core promoter region of the SMN2 gene needs to be identified to study the diverse functional
integrity of this region. For this purpose, the computational promoter prediction and core
promoter prediction analysis was performed and the results of this analysis along with the
previous reported literature were used to propose a model of the core promoter region of the
SMN2 gene. To analyze this model biochemically, the entire Pro region of 720 bp was divided
into five regions (Pro 720, Pro 500, Pro 400, Pro 200 and Pro 100). These five regions were PCR
amplified and were cloned into pTOPO-2.1 e cloning vector. After confirming the insert and the
vector backbone by PCR, restriction analysis and DNA sequencing, the Pro regions were subcloned
into pGL4.74 reporter vector by high throughput directional cloning using EcoRI and
HindWl restriction sites to be transfected directly into HeLa cell lines. Before the transfection the
optimization of the concentration of the test vector pGL4.74 and the control vector pGL4.10 was
performed beside the optimum concentration of test to control vectors for the co-transfection.
The transfected HeLa cell lines were subjected to lysis followed by the dual luciferase assay.
After the normalization of data of the luciferase assay, it was found that the fold change in the
mean percent activity of luciferase (from 1.03 to 2.71) between Pro constructs highly suggested
the role of newly identified computational TSS (-557 bp) in TATA (-411 bp) specific CREB
induced expression of downstream reporter gene through structural and functional role of CRE-II
element. Since, there was no variation identified in the promoter region of the SMN2 gene in the
SMA patients from different clinical severities, it showed that the promoter region of the SMN2 gene has no role in circumscribing the clinical severity of SMA
Recent facts of eating habits and obesity among adolescent; a case of Pakistan
Background: Obesity is an escalating problem that is reaching to pandemic level. Multiple factors may involve in causing obesity such as improper food pattern of physical activities, social and ecological variables, choice of menu and other biological factors. Conducting to a study to evaluate the primary cause. However, a few studies are conducting to see the impact of eating patterns on health and weight. Methods: University students (n=150, ages 18-24 years) 50% males and 50% females were selected for data collection via questionnaire. The outcomes showed that 70 individuals prefer to eat saturated fats that can lead to accumulation of bad cholesterol. 5% females and 5% males prefer using trans-fat that is even worse. On the other hand, 44 respondents prefer to choose low fat food. Results: About 25% individuals are unaware of nutritional on facts of the products but females are more conscious as compared to the men. 55% individuals eat unconsciously while watching television and consume more than the requirement. 94 individuals got attracted by advertisement tactics of food companies and but to eat them. It is also witnessed that males (32%) consume more carbonated drinks than females (13%). 64 students strongly agreed that supplements lead to obesity. In our sample population 10% obese, 14% were overweight and 47% were of normal weight. Conclusion: The major reason of obesity could be that they are eating out more often. It could be due to the fact they are dependent on high calorie food. There is a lack of vegetables and fruits in their diet. On the other hand, fruits, veggies and whole grains are linked to less gain and even weight loss. Making smart food choices can help you stay slim and healthy.
Validation of Vis-Screen Mobile Application for Easy Vision Test by General Public: A pilot study
Visual impairment and blindness is a serious global concern. Poor awareness and late detection are significant factors contributing to the vast number of visually impaired people worldwide. The ubiquitous use of smartphone devices enables the general public to access various services provided easily. Our Vis-Screen mobile application is developed based on the necessity for early detection of any vision problem to avoid further vision loss. Simple built-in algorithm and user-friendly features of this application suits any laypersons to do vision testing among themselves with only minimal training needed, therefore promoting the importance of healthy vision within the community.Keywords: visual acuity; mobile application; vision screening; blindnesseISSN: 2398-4287 © 2019. The Authors. Published for AMER ABRA cE-Bs by e-International Publishing House, Ltd., UK. This is an open-access article under the CC BYNC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Peer–review under responsibility of AMER (Association of Malaysian Environment-Behaviour Researchers), ABRA (Association of Behavioural Researchers on Asians) and cE-Bs (Centre for Environment-Behaviour Studies), Faculty of Architecture, Planning & Surveying, Universiti Teknologi MARA, Malaysia.DOI: https://doi.org/10.21834/e-bpj.v4i12.189
The genetics associated with Primary Congenital Glaucoma
Glaucoma is a progressive optic neuropathy; increased intraocular pressure (IOP) is a modifiable risk factor for primary congenital glaucoma (PCG). Increase IOP causes retinal and optic nerve compression and leads to gradual and irreversible loss of eyesight if left untreated. It is the second most leading cause of blindness. PCG mainly affects children up to the age of three years, and symptoms include epiphora, photalgia, swollen eyes, opaque corneas, blepharospasm, rupture in the retina and ocular nerve damage due to IOP. Early detection, management, and treatment are the keys to preventing vision loss from glaucoma. Many mutations have been discovered in Cytochrome P450 1B1 (CYP1B1) gene to be responsible for causing PCG, and there are still a lot of mutations to be discovered. In this review, we will discuss the genetic aspects of PCG and the most frequent mutations responsible for PCG in Pakistani children. PCG can be handled by decreasing IOP either by medication or by surgery. Genetic counselling plays a significant role in the establishment of proper management of PCG.Keywords: Primary Congenital Glaucoma; IOP; Cyp1b1; Mutation
Intestinal colonization against Vibrio cholerae: host and microbial resistance mechanisms
Vibrio cholerae is a non-invasive enteric pathogen known to cause a major public health problem called cholera. The pathogen inhabits the aquatic environment while outside the human host, it is transmitted into the host easily through ingesting contaminated food and water containing the vibrios, thus causing diarrhoea and vomiting. V. cholerae must resist several layers of colonization resistance mechanisms derived from the host or the gut commensals to successfully survive, grow, and colonize the distal intestinal epithelium, thus causing an infection. The colonization resistance mechanisms derived from the host are not specific to V. cholerae but to all invading pathogens. However, some of the gut commensal-derived colonization resistance may be more specific to the pathogen, making it more challenging to overcome. Consequently, the pathogen has evolved well-coordinated mechanisms that sense and utilize the anti-colonization factors to modulate events that promote its survival and colonization in the gut. This review is aimed at discussing how V. cholerae interacts and resists both host- and microbe-specific colonization resistance mechanisms to cause infection
Immune-Cell-Mediated Cancer Treatment: Advantages, Drawbacks And Future Direction
Cancer ranks as the most lethal and prevalent non-communicable disease in clinical settings. Therapeutic options for cancer comprise chemotherapy, radiotherapy, surgery, and combined treatment. Cancer remission and relapse cases are widespread despite having various advanced medications and sophisticated dissection techniques. A new approach involving immune-cell-mediated cancer therapy has been adopted extensively for cancer treatments by utilizing immune cells. Immunotherapy has gained much attention to prevent and treat various types of cancer. Immunotherapy treatments operate in multiple contexts. Several immunotherapy therapeutic interventions assist the immune function in halting or reducing the advancement of cancer cells. Many also facilitate the immune cells in destroying cancerous cells or safeguarding against cancer from disseminating to certain other regions of the human body. Among other methods, genetic manipulation of immune cells offers hope for innovative anticancer treatment. T lymphocytes and natural killer cells have become the most extensively documented immune cells for immunotherapy. Chimeric antigen receptor T-cell therapy exhibits the most promising blood cancer treatment. However, adoptive NK cell transfer therapy displays potential anticancer treatment options, although more research is needed to be carried out. In addition, cytokine-induced immunomodulation is also plausible for cancer immunotherapy. This review will highlight the most comprehensive information, observations, and consequences associated with different cancer immunotherapy initiatives
Impact of Hypoxia on Astrocyte Induced Pathogenesis
Astrocytes are the most abundant cells of the central nervous system. These cells are of diverse types based on their function and structure. Astrocyte activation is linked mainly with microbial infections, but long-term activation can lead to neurological impairment. Astrocytes play a significant role in neuro-inflammation by activating pro-inflammatory pathways. Activation of interleukins and cytokines causes neuroinflammation resulting in many neurodegenerative disorders such as stroke, growth of tumours, and Alzheimer’s. Inflammation of the brain hinders neural circulation and compromises blood flow by affecting the blood–brain barrier. So the oxygen concentration is lowered, causing brain hypoxia. Hypoxia leads to the activation of nuclear factor kappa B (NFkB) and hypoxia-inducible factors (HIF), which aggravates the inflammatory state of the brain. Hypoxia evoked changes in the blood–brain barrier, further complicating astrocyte-induced pathogenesis
Narrative Review: Use of Competent Stimulating Peptide in Gene Transfer Via Suicide Plasmid in Streptococcus pneumoniae
Natural competency for genetic transformation of Streptococcus pneumoniae causes the emergence of novel or non-vaccine preventable pneumococcal serotypes. This phenomenon has become a global concern as it can spread quickly in the population through inhalation and close contact. The colonisation of S. pneumoniae at the upper respiratory tract can either become commensal or pathogenic. Once the bacterium invades into the body system, it will secrete its toxin and virulence protein to facilitate the invasion. Besides, S. pneumoniae can undergo natural biological transformation via uptake of exogenous DNA by horizontal gene transfer for integration and recombination of the genome. S. pneumoniae natural transformation is aided by competence-stimulating peptide (CSP) that induces the competence of bacteria. Natural transformation cascade of S. pneumoniae via CSP is triggered in the presence of conserved 17-amino acids peptide which is regulated and encoded by comC, comD and comE operon, where comC is responsible in secreting precursor CSP. Nowadays, researchers transforming S. pneumoniae by inserting the mutated S. pneumoniae gene through a vector, suicide plasmid. Suicide plasmids such as pID701, pAUL-A and pVA891 can be transferred but cannot replicate in the bacteria. Homologous recombination process occurs once the mutated gene of suicide plasmid is integrated with wild-type S. pneumoniae. Previous studies had used the transformation of suicide plasmid into S. pneumoniae as it can integrate with host DNA at specific insert for gene transfer. But there is no evidence on the role of CSP in horizontal/gene replacement via suicide plasmid in Streptococcus pneumoniae. This narrative review's scope as per defined purpose statement is to relate and recommend the use of competent stimulating peptide in efficient horizontal gene transfer via suicide plasmids in Streptococcus pneumoniae.Keywords: Streptococcus pneumoniae transformation; CSP-based transformation; Gene transfer via suicide plasmid; Genetic exchange; Competence-stimulating peptid
Detection, quantification and genotype distribution of HCV patients in Lahore, Pakistan by real-time PCR
Background: Hepatitis C virus (HCV) is considered as \u201cViral Time
Bomb\u201d suggested by the World Health Organization and if it is not
treated timely, it will lead towards cirrhosis and hepatocellular
carcinoma (HCC). Objective: The purpose of the present research is to
study possible risk factors, frequent genotypes of HCV and its
association with different age groups. Methods: Suspected blood samples
from HCV patients were collected from different hospitals of Lahore,
Pakistan. Out of 1000 HCV suspected samples, 920 samples were found HCV
positive detected by Anti-HCV ELISA, CobasR. kit. The quantification of
HCV load was determined by HCV quantification kit and LINEAR ARRAY KIT
(Roche) was used for genotype determination by Real-Time PCR (ABI).
Statistical analysis was done by using Microsoft Excel. Results: Out of
920 subjects, 77 subjects (8.4%) were false positive and they were not
detected by nested PCR. Three PCR positive samples were untypeable.
Genotype 3 was predominant in Lahore which was 83.5%, whereas type 1
and 2 were 5.1% and 0.7% respectively. There were also mixed genotypes
detected, 1 and 3 were 0.4%, 2 and 3 were 1.41% and 3 and 4 were 0.2%
only. Male were more infected of HCV in the age <40 years and
females >40years. Conclusion: The major risk factor for HCV
transmission is by use of unsterilized razors/blades. It is necessary
to spread awareness among the general population of Pakistan about HCV
transmission risk factors. Regular physical examination at least once a
year is recommended, so that early detection of HCV could be done
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