Institute of Biochemistry and Biotechnology

Lahore, Pakistan

Institute of Biochemistry and Biotechnology

Lahore, Pakistan
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Burstenbinder K.,Leibniz Institute of Plant Biochemistry | Moller B.,Institute of Computer Science | Plotner R.,Leibniz Institute of Plant Biochemistry | Stamm G.,Leibniz Institute of Plant Biochemistry | And 6 more authors.
Plant Physiology | Year: 2017

Calcium (Ca2+) signaling and dynamic reorganization of the cytoskeleton are essential processes for the coordination and control of plant cell shape and cell growth. Calmodulin (CaM) and closely related calmodulin-like (CML) polypeptides are principal sensors of Ca2+ signals. CaM/CMLs decode and relay information encrypted by the second messenger via differential interactions with a wide spectrum of targets to modulate their diverse biochemical activities. The plant-specific IQ67 DOMAIN (IQD) family emerged as possibly the largest class of CaM-interacting proteins with undefined molecular functions and biological roles. Here, we show that the 33 members of the IQD family in Arabidopsis (Arabidopsis thaliana) differentially localize, using green fluorescent protein (GFP)-tagged proteins, to multiple and distinct subcellular sites, including microtubule (MT) arrays, plasma membrane subdomains, and nuclear compartments. Intriguingly, the various IQD-specific localization patterns coincide with the subcellular patterns of IQD-dependent recruitment of CaM, suggesting that the diverse IQD members sequester Ca2+-CaM signaling modules to specific subcellular sites for precise regulation of Ca2+-dependent processes. Because MT localization is a hallmark of most IQD family members, we quantitatively analyzed GFP-labeled MT arrays in Nicotiana benthamiana cells transiently expressing GFP-IQD fusions and observed IQD-specific MT patterns, which point to a role of IQDs in MT organization and dynamics. Indeed, stable overexpression of select IQD proteins in Arabidopsis altered cellular MT orientation, cell shape, and organ morphology. Because IQDs share biochemical properties with scaffold proteins, we propose that IQD families provide an assortment of platform proteins for integrating CaM-dependent Ca2+ signaling at multiple cellular sites to regulate cell function, shape, and growth. © 2017 American Society of Plant Biologists. All rights reserved.

Afridi S.Q.,Quality Operations Laboratory | Ali M.M.,Institute of Biochemistry and Biotechnology | Zahid M.N.,Quality Operations Laboratory | Afridi I.Q.,Ideal Laboratories | And 2 more authors.
Virology Journal | Year: 2014

Background: Hepatitis C virus (HCV) is highly infectious pathogen which is responsible for causing Hepatitis around 200 million individuals worldwide. In Pakistan, 4.7% of HCV prevalence has been reported and HCV genotype 3a has been found to be the major source of infection in Pakistan but still there is lack of information on distribution of HCV genotypes and viral load in various geographical regions of Pakistan. Therefore, current study was designed to determine distribution of HCV genotypes as well viral load in different areas of Punjab province of Pakistan. Findings. A total of 995 serum samples were taken from those individuals in which antibodies against HCV were detected through ELISA, from different regions of Punjab i.e. Lahore 317(31.85%), Faisalabad 70(7.03%), Gujranwala 129(12.96%), Gujrat 106(10.65%), Sialkot 94(9.44%), Sargodha 60(6.03%), Mandibaha-ud-din 135(13.56%), Jhang 86(8.64%). Qualitative PCR was performed to determine viral load and genotyping was performed using Nested PCR. Chi-square test was used to determine the age and sex-wise prevalence of HCV. Out of 995 samples, 888 samples were found positive for HCV RNA. In all regions, genotype 3a showed highest prevalence (82.81%) followed by genotype 1 (3.41%), mixed genotypes (2.41%), genotype 2 (0.50%), genotype 5 (0.1%) and unclassified genotypes (10.75%). Viral load in 29.5% patients infected with genotype 3a was less than 600,000 IU/mL, while it was between 600,000-800,000 IU/mL in 27.9% patients and 25.22% patients had more than 800,000 IU/mL viral load. Conclusion: HCV genotype 3a is the most prevalent genotype in various regions of Punjab. Viral load of HCV patients in these different regions of Punjab are reported for the first time. Moreover, based upon these results the Patients having viral load below 800,000 IU/mL would be expected to show better response of anti-HCV therapy. © 2014 Afridi et al.; licensee BioMed Central Ltd.

Tahir M.S.,Institute of Biochemistry and Biotechnology | Hussain T.,The University of Lahore | Babar M.E.,The University of Lahore | Nadeem A.,Institute of Biochemistry and Biotechnology | And 4 more authors.
Journal of Animal and Plant Sciences | Year: 2015

A molecular genetics tool comprised of a panel of 15 microsatellite markers was developed and used to investigate parentage and breed characterization of two most kept breeds of dogs including German shepherd and Labrador retriever in Pakistan. Blood samples of 20 dog families (10 from each breed) were collected from Army dog Breeding Training Center and School, Rawalpindi, Pakistan and Kennel Club of Pakistan. Genomic DNA was extracted by standard inorganic protocol. Microsatellite markers with high Polymorphism Information Content (PIC) and He (Hetrozygosity) values were selected and optimized into four multiplexes. Amplification reactions were followed by genotyping in 7% non-denaturing polyacrylamide gel electrophoresis (PAGE). Parentage analysis of 20 families using this panel of microsatellite markers was 100% successful. Average values of Polymorphism Information Content (PIC), Heterozygosity (He) and Combined Power of Exclusion (CPE) combined for both of the breeds were found to be 0.724, 0.6345 and 0.9998 respectively. Moreover, deviation from Hardy-Weinberg equilibrium equation was observed moderately for both dog breeds. Allelic frequencies for majority of the microsatellite markers between both dog breeds were clearly distinct. This study demonstrated the panel of 15 microsatellite markers could effectively validate parentage and breed characterization in dogs. © 2015, Pakistan Agricultural Scientists Forum. All rights reserved.

Ahmad A.,BUITEMS | Ahmad A.,Institute of Biochemistry and Biotechnology | Daud S.,University of Punjab | Kakar N.,BUITEMS | And 9 more authors.
Molecular Vision | Year: 2011

Purpose: To determine the cause of Leber congenital amaurosis (LCA) and developmental cataracts in a consanguineous Pakistani family. Methods: The diagnosis was established in all affected individuals of a Pakistani LCA family by medical history, funduscopy, and standard ERG. We performed genome-wide linkage analysis for mapping the disease locus in this family. Results: Congenitally severely reduced visual acuity and nystagmus were reported for all patients who, in the later phase of the disease, also developed cataracts. LCA in the family cosegregated with homozygosity for a single nucleotide polymorphism (SNP) haplotype on chromosome 6p14.1. The respective candidate region contained Leber congenital amaurosis 5 (LCA5), a gene previously reported to underlie LCA. We subsequently identified a novel truncating mutation in exon 4 of LCA5, c.642delC, in homozygous state in all affected persons of the family. Conclusions: We report a novel LCA5 mutation causing LCA in a Pakistani family. Developmental cataracts were present in two of the four patients, raising the possibility that LCA5 mutations may predispose to this additional ocular pathology. © 2011 Molecular Vision.

Hussain R.,Buitems | Hussain R.,Sardar Bahadur Khan Women's University | Daud S.,Center for Applied Molecular Biology | Daud S.,Institute of Biochemistry and Biotechnology | And 6 more authors.
Molecular Biology Reports | Year: 2012

Canavan disease (OMIM 271900) is an autosomal recessive lethal neurodegenerative disorder characterized by spongy degeneration of the brain. A highly consanguineous Pakistani family with Canavan disease was enrolled on the basis of diagnosis. All the affected individuals have mental retardation, megalocephaly and degradation of motor skills, poor head control, partial vision loss, weakness of the muscles and raised urinary concentration of N-acetyl aspartic acid in the urine. Blood samples were collected from affected as well as normal siblings and processed for DNA purification. Linkage analysis was performed by typing three short tandem repeat markers D17S1583 (7.19 cM), D17S1828 (10.02 cM) and D17S919 (14.69 cM) for an already-reported gene/locus ASPA at chromosome 17p13.2 causing Canavan disease. During linkage analysis, all the affected individuals were homozygous for short tandem repeat markers while the normal siblings were heterozygous showing co-segregation of the disease. Gene ASPA (NM-000049) was undertaken to sequence for mutation analysis. As a result of sequence analysis, we found missense substitution 740A?G (p.G274R) in exon 6 of gene ASPA. To our knowledge, this is the first report about Canavan disease on a Pakistani family. © Springer Science+Business Media B.V. 2011.

Chen X.,Nanjing Agricultural University | Chen X.,Louisiana State University Health Sciences Center | Gu Y.,Nanjing Agricultural University | Gu Y.,Louisiana State University Health Sciences Center | And 8 more authors.
PLoS ONE | Year: 2014

Maduramicin, a polyether ionophore antibiotic derived from the bacterium Actinomadura yumaensis, is currently used as a feed additive against coccidiosis in poultry worldwide. It has been clinically observed that maduramicin can cause skeletal muscle and heart cell damage, resulting in skeletal muscle degeneration, heart failure, and even death in animals and humans, if improperly used. However, the mechanism of its toxic action in myoblasts is not well understood. Using mouse myoblasts (C2C12) and human rhabdomyosarcoma (RD and Rh30) cells as an experimental model for myoblasts, here we found that maduramicin inhibited cell proliferation and induced cell death in a concentration-dependent manner. Further studies revealed that maduramicin induced accumulation of the cells at G0/G1 phase of the cell cycle, and induced apoptosis in the cells. Concurrently, maduramicin downregulated protein expression of cyclin D1, cyclin-dependent kinases (CDK4 and CDK6), and CDC25A, and upregulated expression of the CDK inhibitors (p21Cip1 and p27 Kip1), resulting in decreased phosphorylation of Rb. Maduramicin also induced expression of BAK, BAD, DR4, TRADD and TRAIL, leading to activation of caspases 8, 9 and 3 as well as cleavage of poly ADP ribose polymerase (PARP). Taken together, our results suggest that maduramicin executes its toxicity in myoblasts at least by inhibiting cell proliferation and inducing apoptotic cell death.Copyright: © 2014.

Sherwani S.K.,University of Karachi | Ahmad H.,University of Queensland | Ahmad T.,Hazara University | Hussain T.,Institute of Biochemistry and Biotechnology | And 3 more authors.
World Journal of Medical Sciences | Year: 2014

The current study was carried out on 550 healthy population having 250 males and 300 females. Five ml venousblood was collected following standard biosafety measures. ABO blood grouping was done by Tile method and found blood group B dominant in both sexes i.e 180 male and 120 female were found with blood group B. Moreover, 2 ml of saliva was also collected from allvolunteers. Secretor status was detected from the saliva byhaemagglutination inhibition method and found 278 female and 234 male were secretors. © IDOSI Publications, 2014.

Kolenko P.,Institute of Biochemistry and Biotechnology | Kolenko P.,Czech Institute of Macromolecular Chemical | Koch B.,Probiodrug | Rahfeld J.-U.,Probiodrug | And 3 more authors.
Acta Crystallographica Section F: Structural Biology and Crystallization Communications | Year: 2013

The structure of ligand-free glutaminyl cyclase (QC) from Drosophila melanogaster (DmQC) has been determined in a novel crystal form. The protein crystallized in space group I4, with unit-cell parameters a = b = 122.3, c = 72.7 Å. The crystal diffracted to a resolution of 2 Å at the home source. The structure was solved by molecular replacement and was refined to an R factor of 0.169. DmQC exhibits a typical /β-hydrolase fold. The electron density of three monosaccharides could be localized. The accessibility of the active site will facilitate structural studies of novel inhibitor-binding modes. © 2013 International Union of Crystallography.

Chu S.-C.,Central Taiwan University of Science and Technology | Yu C.-C.,Institute of Oral Science | Yu C.-C.,Chung Shan Medical University | Hsu L.-S.,Institute of Biochemistry and Biotechnology | And 5 more authors.
Molecular Pharmacology | Year: 2014

Metastasis is the most common cause of cancer-related death in patients, and epithelial-to-mesenchymal transition (EMT) is essential for cancer metastasis, which is a multistep complicated process that includes local invasion, intravasation, extravasation, and proliferation at distant sites. When cancer cells metastasize, angiogenesis is also required for metastatic dissemination, given that an increase in vascular density will allow easier access of tumor cells to circulation, and represents a rational target for therapeutic intervention. Berberine has several anti-inflammation and anticancer biologic effects. In this study, we provided molecular evidence that is associated with the antimetastatic effect of berberine by showing a nearly complete inhibition on invasion (P> 0.001) of highly metastatic SiHa cells via reduced transcriptional activities of matrix metalloproteinase-2 and urokinase-type plasminogen activator. Berberine reversed transforming growth factor-β1-induced EMT and caused upregulation of epithelial markers such as E-cadherin and inhibited mesenchymal markers such as N-cadherin and snail-1. Selective snail-1 inhibition by snail-1-specific small interfering RNA also showed increased E-cadherin expression in SiHa cells. Berberine also reduced tumor-induced angiogenesis in vitro and in vivo. Importantly, an in vivo BALB/c nude mice xenograft model and tail vein injection model showed that berberine treatment reduced tumor growth and lung metastasis by oral gavage, respectively. Taken together, these findings suggested that berberine could reduce metastasis and angiogenesis of cervical cancer cells, thereby constituting an adjuvant treatment of metastasis control. Copyright © 2014 by The American Society for Pharmacology and Experimental Therapeutics.

PubMed | Institute of Biochemistry and Biotechnology
Type: Journal Article | Journal: Genes, brain, and behavior | Year: 2014

Hypothalamic neuropeptides, including neuropeptide Y (NPY) and proopiomelanocortin (POMC), have been found to control the appetite-suppressing effect of amphetamine (AMPH). In this study, we have examined whether dopamine receptor (DAR), phosphatidylinositol 3-kinase (PI3K) and nuclear factor-kappaB (NF-B) are involved in AMPHs action. We administered AMPH to rats once a day for 4days and assessed and compared changes in hypothalamic NPY, melanocortin receptor 4 (MC4R), PI3K, pAkt and NF-B expression. We found that the inhibition of DAR increased NPY, but decreased MC4R, PI3K and NF-B expression, compared with AMPH-treated rats. Moreover, MC4R, PI3K, pAkt and NF-B increased with the maximum response on Day 2, which was consistent with the response of feeding behavior, but was opposite to the expression of NPY. Furthermore, we found that the intracerebroventricular infusion of the PI3K inhibitor or NF-B antisense could attenuate AMPH-induced anorexia, and partially reverse the expression of NPY, MC4R, PI3K, Akt and NF-B back toward a normal level. We, therefore, suggest that DAR-PI3K-NF-B signaling in the hypothalamus plays functional roles in the modulation of NPY and POMC neurotransmissions and in the control of AMPH-evoked appetite suppression.

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