Abid Azhar

Dental caries in diabetes mellitus: role of salivary flow rate and minerals

This study was designed to evaluate the possible protective role of salivary factors like salivary flow rate and adequate level of calcium, phosphate, and fluoride in diabetes mellitus type 2 patients with dental caries. A total of 398 diabetes mellitus type 2 patients with dental caries and 395 age- and sex-matched non-diabetic subjects with dental caries were included as controls, all of whom gave informed consent. All subjects were divided into four groups according to their age. Decayed, missed, and filled teeth (DMFT) were scored to indicate the severity of dental caries. Saliva was collected, flow rate was noted, and calcium, phosphate, and fluoride were analyzed. The blood glucose, HbA1c, and DMFT indices were found to be significantly high in diabetic patients as compared to controls. The salivary flow rate, calcium, phosphate, and fluoride were found to be significantly low whereas no significant difference was found in salivary magnesium in patients as compared to controls. Optimum salivary flow rate is responsible for establishing protective environment against dental caries. Adequate level of salivary calcium, phosphate, and fluoride is also involved in significant deposition of these minerals in plaque, which greatly reduces the development of caries in the adjacent enamel of teeth.

Protective Effects of Salivary Factors in Dental Caries in Diabetic Patients of Pakistan

Salivary factors have been studied for their effects on the process of dental caries in patients of diabetes mellitus type 2. In this study, protective role of salivary pH, salivary flow rate, and salivary calcium is assessed in the patients of diabetes mellitus type 2 with dental caries. The samples of saliva were collected from 400 patients of diabetes mellitus type 2 and 300 age- and sex- matched controls after getting informed consent. All the subjects were classified into four groups according to age. The severity of dental caries was counted by decayed, missed, and filled teeth (DMFT) score. The salivary pH, flow rate, and calcium levels were found to be low in patients as compared to controls. The levels of fasting blood sugar, HbA1c, and DMFT score were found to be significantly high in patients than controls. The glycemic factors were significantly correlated with salivary factors indicating their influence on progression of caries in diabetes. On the basis of findings, it is concluded that the suitable salivary pH and flow rate may be regarded as main protective factors against dental caries in diabetes. Optimum level of salivary calcium may be responsible for continuous supply of calcium to arrest the demineralization and help reduce the occurrence of dental caries.

Estimation of serum osteocalcin and telopeptide-C in postmenopausal osteoporotic females

SummaryA negative correlation of serum C-terminal telopeptide of type I collagen (telopeptide-C) was observed with bone mass density (BMD) in postmenopausal control (r = −0.70, p < 0.05) and osteoporotic females (r = −0.46, p < 0.05) indicating increased bone resorption in these subjects. Since telopeptide-C is a significant determinant of bone loss, it can be used, in combination with bone mass measurement, for the assessment of postmenopausal females.IntroductionThe aim of this study was to find out the significance of serum osteocalcin, a marker of bone formation, and C-terminal telopeptide of type I collagen, a marker of bone resorption, in evaluating osteoporotic patients and to find out their relationship with bone mass density.MethodsOne hundred and fifty (150) females; 50 premenopausal (age=31.13 ± 1.29), 50 postmenopausal (age = 54.36 ± 0.81) and 50 postmenopausal osteoporotic patients (age = 58.6 ± 0.701) were included in this study. The postmenopusal subjects, with and without osteoporosis, were selected from different osteoporotic clinics and healthy premenopausal females were selected from general population. Height, weight, BMI, waist/hip ratio, age at menarche, years since menopause in case of postmenopausal women, history of disease, and fracture, if any, were recorded. BMD assessment was done on calcaneous by peripheral ultrasound bone densitometery on Sahara Clinical Bone Sonometer and T-scores were calculated. Serum levels of osteocalcin and C-terminal telopeptide of type I collagen and calcium were measured.ResultsA lower BMD in postmenopausal subjects, with and without osteoporosis (p < 0.001), indicating increased bone loss with aging and menopause, was observed. A negative correlation was found between age and BMD (r = −0.67, p < 0.05). No correlation was found between osteocalcin and BMD among these groups, suggesting heterogeneity of osteocalcin fragments in serum that limits its significance in the evaluation of osteoporosis. A positive correlation was found between osteocalcin and telopeptide-C (r = 0.26, p < 0.05). A positive correlation of telopeptide-C with age (r = 0.45, p < 0.05) and a negative correlation with BMD (r = −0.46, p < 0.05) was observed indicating increased bone resorption in postmenopausal control and postmenopausal osteoporotic patients.ConclusionC-terminal telopeptide of type I collagen appears to be a significant determinant of bone loss and may be used as a valuable tool in the assessment of postmenopausal osteoporotic patients.

Genetic variants of ACE (Insertion/Deletion) and AGT (M268T) genes in patients with diabetes and nephropathy

Introduction: Diabetes mellitus (DM) has been a growing epidemic worldwide and poses a major socio-economic challenge. The leading cause of DM death is nephropathy due to end-stage renal disease (ESRD). This study aims to identify the possible association of I/D variants of the ACE gene and M268T (rs699) of the AGT gene of renin–angiotensin–aldosterone system (RAAS). Materials and methods: Study subjects include 115 patients with DM, 110 with diabetic nephropathy (DN) and 110 controls. Fasting blood samples were collected for biochemical analyses and PCR amplification of specific regions of the ACE and AGT genes using primers. Results: The distribution of ACE (I/D) II 28.8%, ID 35.6% and DD 35.6% while in DN II 24.5%, ID 41% and DD 34.5%. The AGT (M268T) genotypes were distributed in DM as TT 30.4%, MT 66.9% and MM 2.6% while in DN subjects TT 56.4%, MT 42.7% and MM 0.9%. Conclusion: Significant differences were observed in the DD genotype and D allele of the ACE gene and the TT genotype and T allele of AGT genes between diabetic patients with and without nephropathy. The study may conclude that the D allele polymorphism in the ACE gene and the T allele polymorphism in AGT gene may be considered as genetic risk factors for the development of nephropathy in diabetes.

Protein expression profiling of nuclear membrane protein reveals potential biomarker of human hepatocellular carcinoma

BackgroundComplex molecular events lead to development and progression of liver cirrhosis to HCC. Differentially expressed nuclear membrane associated proteins are responsible for the functional and structural alteration during the progression from cirrhosis to carcinoma. Although alterations/ post translational modifications in protein expression have been extensively quantified, complementary analysis of nuclear membrane proteome changes have been limited. Deciphering the molecular mechanism that differentiate between normal and disease state may lead to identification of biomarkers for carcinoma.ResultsMany proteins displayed differential expression when nuclear membrane proteome of hepatocellular carcinoma (HCC), fibrotic liver, and HepG2 cell line were assessed using 2-DE and ESI-Q-TOF MS/MS. From the down regulated set in HCC, we have identified for the first time a 15 KDa cytochrome b5A (CYB5A), ATP synthase subunit delta (ATPD) and Hemoglobin subunit beta (HBB) with 11, 5 and 22 peptide matches respectively. Furthermore, nitrosylation studies with S-nitrosocysteine followed by immunoblotting with anti SNO-cysteine demonstrated a novel and biologically relevant post translational modification of thiols of CYB5A in HCC specimens only. Immunofluorescence images demonstrated increased protein S-nitrosylation signals in the tumor cells and fibrotic region of HCC tissues. The two other nuclear membrane proteins which were only found to be nitrosylated in case of HCC were up regulated ATP synthase subunit beta (ATPB) and down regulated HBB. The decrease in expression of CYB5A in HCC suggests their possible role in disease progression. Further insight of the functional association of the identified proteins was obtained through KEGG/ REACTOME pathway analysis databases. String 8.3 interaction network shows strong interactions with proteins at high confidence score, which is helpful in characterization of functional abnormalities that may be a causative factor of liver pathology.ConclusionThese findings may have broader implications for understanding the mechanism of development of carcinoma. However, large scale studies will be required for further verification of their critical role in development and progression of HCC.

P53 (Pro72Arg) polymorphism associated with the risk of oral squamous cell carcinoma in gutka, niswar and manpuri addicted patients of Pakistan

OBJECTIVES The chewing habit of paan, chhaliya, and tobacco is common in the traditional culture of Pakistan. Currently, niswar, gutka and manpuri are also commercially available in the Pakistani market. Epidemiologic evidences and increased rate of oral squamous cell carcinoma (OSCC) cases may indicate a direct relationship of these chewing habits with oral carcinogenesis. The p53 gene has been known to be a tumor suppressor gene that is found mutated in common human cancers. The p53 gene contains a single nucleotide polymorphism at codon 72 of exon 4 which encodes either proline (Pro) or arginine (Arg). The aim of the present study was to investigate association of p53 gene codon 72 polymorphism with patients of oral squamous cell carcinoma consuming these carcinogenic chewable materials. MATERIALS AND METHODS Blood and tissue samples of 260 OSCC patients were collected with informed consent from the local hospitals of Karachi. The patients were compared with controls of similar age and sex. The exon 4 of p53 gene was examined by PCR-SSCP. The tumor samples showing mobility shift were purified and sequenced. RESULTS The C>G missense mutation at nucleotide position 215 of the coding sequence was identified which substitutes proline with arginine at codon 72 of p53 protein. When the data for CCC72CGC polymorphism was analyzed statistically, a significant difference was observed between OSCC and control samples. The Pro allelic frequencies were significantly higher in OSCC patients as compare to controls. The current study indicated the Pro form of p53 codon 72 increases the risk of developing OSCC in Pakistani population. The risk ratio for Pro allele was 1.5004 (95% confidence interval: 1.2559 to 1.7924) and odds ratio of Pro allele was 2.389 (95% confidence interval: 1.5591 to 2.8137) in comparison with the Arg and Pro alleles in the OSCC group. CONCLUSION These evidences suggest that there may be specific genetic targets with these chewing ingredients that are responsible for causing OSCC. The p53 codon 72 polymorphism is associated with OSCC at somatic cell level but the polymorphism was not associated at inherited level.

Phylogenetic analysis of HDV isolates from HBsAg positive patients in Karachi, Pakistan.

BackgroundIn spite of a high occurrence of Hepatitis Delta in the province of Sindh in Pakistan, no genetic study of Hepatitis Delta virus (HDV) isolates from this region was carried out. The aim of this study is to analyze the genetic proximity within local HDV strains, and relationship with other clades of HDV, using phylogenetic analysis.ResultsPhylogenetic analysis of nucleotide sequences of the Hepatitis Delta Antigen (HDAg) R0 region obtained in this study, showed considerable diversity among the local strains with a potential subgroup formation within clade I. The multiple sequence alignment of predicted amino acids within clade I showed many uncommon amino acid substitutions within some conserved regions that are crucial for replication and assembly of HDV.ConclusionsThe studied strains showed a range of genetic diversity within HDV clade I. There is clustering of sequences into more than one group, along with formation of potential subgroup within clade I. Clustering shows the genetic closeness of strains and indicates a common origin of spread of HDV infection. Further phylogeny-based studies may provide more information about subgroup formation within clade I and may be used as an effective tool in checking and/or preventing the spread of hepatitis D virus infection in this region.

Regeneration in sugarcane via somatic embryogenesis and genomic instability in regenerated plants

In the present study, embryogenic calli of sugarcane variety BL4 were induced using 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin in different concentrations and combinations. In contrast to earlier studies, embryogenic callus sectors were identified and isolated microscopically within 1–2 weeks. Subsequently, 51 media formulations were used for regeneration of proliferated embryogenic callus, using MS medium supplemented with three different cytokinins [kinetin, 6-Benzylamino purine (BAP), and thidiazuron (TDZ)] and auxins (IAA/NAA and IBA) in different combination and concentrations. After acclimatization, the genomic DNA of regenerated plants was studied to explore the insertion polymorphism of transposable elements in order to ascertain the variation among somaclones. Though low concentration of kinetin with 2,4-D was found supportive to embryogenic callus development, the highest number of regenerated plantlets was observed using BAP (1 μM), however the plantlets had very low fresh weight (2.2 g). Conversely, TDZ alone supported a significant increase in the number of plantlets regenerated (38–40) with higher fresh weight. The somaclones generated during this study showed considerable positional polymorphism of activator-like transposable elements possibly due to the stress associated with in vitro culture. This study provides a procedure to produce regenerated sugarcane plants from embryogenic callus in a relatively short time.

SLC11A1 polymorphisms and host susceptibility to cutaneous leishmaniasis in Pakistan

BackgroundThe vector-borne cutaneous leishmaniasis (CL) is endemic in several regions of Pakistan mainly affecting poor populations. Host genetic factors, particularly SLC11A1 (solute carrier transmembrane protein) within macrophages, play a crucial role in disease pathology and susceptibility. Association of SLC11A1 with cutaneous leishmaniasis, a neglected tropical disease, is not well established. Inconsistencies have been observed within different populations worldwide with respect to genetic susceptibility. This study was designed to investigate genetic variation(s) in SLC11A1 and to assess possible association with cutaneous leishmaniasis in Pakistan.ResultsEight polymorphisms (rs2276631, rs3731864, rs2290708, rs2695342, rs201565523, rs17215556, rs17235409, rs17235416) were genotyped across SLC11A1 in 274 patients and 119 healthy controls. Six polymorphisms were studied by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and sequencing. Two single nucleotide polymorphisms were analyzed with newly designed semi-nested PCR assays. Case-control analysis showed no association between selected polymorphisms in SLC11A1 and cutaneous leishmaniasis. No significant difference was observed in the distribution of alleles between leishmaniasis patients and healthy individuals. Strong pairwise linkage disequilibrium was observed between rs2276631 and rs2290708 (r2 = 64); and rs17235409 and rs17235416 (r2 = 78).ConclusionsThis study shows that genetic variations in the candidate gene SLC11A1 do not affect susceptibility to cutaneous leishmaniasis in the sample population from Pakistan.

Bnsro1: A new homologue of Arabidopsis thaliana rcd1 from Brassica napus

Abstract Generation of reactive oxidation species in response to different types of stress is a general phenomenon observed in plants. It is considered to be a molecular signaling mechanism of plants to encounter adverse effects. Radical-induced cell death 1 (rcd1 or Atrcd1) gene of Arabidopsis thaliana is a stress responsive gene known to interact with several transcription factors during different types of stress. It is predicted to provide scaffold for mediating interactions between two proteins using its WWE and RST domains. It also has an inactive PARP catalytic domain forming the Similar like rcd1 (SRO) family of plant PARPs along with its homologs. In this study a new homolog from Brassica napus genome (Bnsro1) was identified. Analysis of Bnsro1 was done to predict function on computational basis by comparison with its homolog. Bnsro1 has similarities with Atrcd1 at sequence level and contains same globular domains. It is predicted to be catalytically active as it conserves the 16 amino acids required for NAD+ binding pocket. Most of the motifs and sites harbored by AtRCD1 are also conserved in BnSRO1. Both translated proteins have intrinsically disordered regions but BnSRO1 is predicted to be more unstructured than AtRCD1. Therefore, the analysis done in this report predicts Bnsro1 as a new member of SRO family having a comparable structural and functional relationship to the Arabidopsis thaliana rcd1.