Cross-linked pH-sensitive pectin and acrylic acid based hydrogels for controlled delivery of metformin.

The purpose of present study is to load Metformin HCl into pH-sensitive hydrogels to have sustained release over a period of time. The hydrogel was synthesized from naturally occurring polysaccharide pectin and monomer acrylic acid (AA) using ethylene glycol dimethacrylate (EGDMA) as cross-linker under controlled conditions for polymerization at 45°C for one hr, 50°C for two hrs, 55°C for three hrs, 60°C for four hrs and finally 65˚C for 12 hrs. Hydrogels were characterized for dynamic/equilibrium swelling, sol-gel fraction analysis, diffusion coefficient and percentage porosity. Hydrogels were tested by FTIR, XRD and SEM for structure and surface morphology respectively. Experimental in-vitro drug release data was applied to kinetic models. Formation of strong bonding between pectin and AA was supported by FTIR. The intensity of XRD peaks was reduced in non-loaded and loaded hydrogels compared to active drug substance. The non-loaded hydrogel showed discrete porous structure whereas loaded hydrogels were fibrous and smooth. Hydrogels showed higher swelling in the solutions of pH 6.5 and 7.5 as compared to in the solutions of pH 1.2 and 5.5. The diffusion coefficient decreases with the increase of AA and pectin concentrations. It was observed upon increasing the EGDMA concentration porosity decreases. The release of drug from all compositions of hydrogels took place through non-Fickian diffusion mechanism.

Selected reaction monitoring to differentiate and relatively quantitate isomers of sulfated and unsulfated core 1 O-glycans from salivary MUC7 protein in rheumatoid arthritis.

Rheumatoid arthritis is a common and debilitating systemic inflammatory condition affecting up to 1% of the world's population. This study aimed to investigate the immunological significance of O-glycans in chronic arthritis at a local and systemic level. O-Glycans released from synovial glycoproteins during acute and chronic arthritic conditions were compared and immune-reactive glycans identified. The sulfated core 1 O-glycan (Galß1-3GalNAcol) was immune reactive, showing a different isomeric profile in the two conditions. From acute reactive arthritis, three isomers could be sequenced, but in patients with chronic rheumatoid arthritis, only a single 3-Gal sulfate-linked isomer could be identified. The systemic significance of this glycan epitope was investigated using the salivary mucin MUC7 in patients with rheumatoid arthritis and normal controls. To analyze this low abundance glycan, a selected reaction monitoring (SRM) method was developed to differentiate and relatively quantitate the core 1 O-glycan and the sulfated core 1 O-glycan Gal- and GalNAc-linked isomers. The acquisition of highly sensitive full scan linear ion trap MS/MS spectra in addition to quantitative SRM data allowed the 3- and 6-linked Gal isomers to be differentiated. The method was used to relatively quantitate the core 1 glycans from MUC7 to identify any systemic changes in this carbohydrate epitope. A statistically significant increase in sulfation was identified in salivary MUC7 from rheumatoid arthritis patients. This suggests a potential role for this epitope in chronic inflammation. This study was able to develop an SRM approach to specifically identify and relatively quantitate sulfated core 1 isomers and the unsulfated structure. The expansion of this method may afford an avenue for the high throughput investigation of O-glycans.

Hydrogeochemical characteristics, stable isotopes, positive matrix factorization, source apportionment, and health risk of high fluoride groundwater in semiarid region.

Chronic exposure to high fluoride (F-) levels in groundwater causes community fluorosis and non-carcinogenic health concerns in local people. This study described occurrence, dental fluorosis, and origin of high F-groundwater using δ2H and δ18O isotopes at semiarid Gilgit, Pakistan. Therefore, groundwater (n = 85) was collected and analyzed for F- concentrations using ion-chromatography. The lowest F- concentration was 0.4 mg/L and the highest 6.8 mg/L. F- enrichment is linked with higher pH, NaHCO3, NaCl, δ18O, Na+, HCO3-, and depleted Ca+2 aquifers. The depleted δ2H and δ18O values indicated precipitation and higher values represented the evaporation effect. Thermodynamic considerations of fluorite minerals showed undersaturation, revealing that other F-bearing minerals viz. biotite and muscovite were essential in F- enrichment in groundwater. Positive matrix factorization (PMF) and principal component analysis multilinear regression (PCAMLR) models were used to determine four-factor solutions for groundwater contamination. The PMF model results were accurate and reliable compared with those of the PCAMLR model, which compiled the overlapping results. Therefore, 28.3% exceeded the WHO permissible limit of 1.5 mg/L F-. Photomicrographs of granite rocks showed enriched F-bearing minerals that trigger F- in groundwater. The community fluorosis index values were recorded at > 0.6, revealing community fluorosis and unsuitability of groundwater for drinking.

Structural vaccinology-based design of multi-epitopes vaccine against Streptococcus gordonii and validation using molecular modeling and immune simulation approaches.

Streptococcus gordonii is an oral bacterium colonizing the dental cavity and leading to plaque formation. This pervasive colonizer is also the etiologic agent of bacterial endocarditis and has a major role in infective endocarditis. The bacteria reach the heart through oral bleeding, leading to inflammation of cardiovascular valves. Over the past 50 years, it has shown a significant pathogenic role in immunocompromised and neutropenic patients. Since antibiotic resistance has created prophylaxis failure towards infective endocarditis, a potent therapeutic candidate is needed. Therefore, multi-epitopes vaccine offers advantages over the other approaches. Thus, herein, numerous molecular-omics tools were exploited to mine immunogenic peptides, i.e., T-cell and B-cell epitopes, and construct a vaccine sequence. Our findings revealed a total of 24 epitopes, including CTL, HTL, and B-cell are responsible for imparting immune responses, which were combined with the help of different linkers, and MEVC was constructed. Multifactorial validation of the candidate vaccine was performed to minimize the risk factors. The final sequence was docked with TLR2 to validate its conformation compatibility with receptor and long-term interactions stability. Our analysis revealed that the vaccine construct is immunogenic and non-allergenic. The construct also established various contacts with the immune receptor. Finally, the vaccine sequence was reverse-translated, optimized for codon usage, and analyzed for expression in the Escherichia coli K12 strain. Maximum expression was noted with a CAI score of 0.95. In silico immune simulation revealed that the antigen was neutralized on the 3rd day after injection. In conclusion, the current study warrants validation of the vaccine construct both in in vitro and in vivo models for accurate therapeutic intervention.

An overview of treatment response rates to various anti-viral drugs in Pakistani hepatitis B virus infected patients.

Hepatitis B virus (HBV) is one of the leading health problem with up to 350 million affected people worldwide including 4.5 million only in Pakistan. It has mortality rate of 0.5 to 1.2 million per year worldwide. Pakistan lies in the endemic region with 3-5% HBV carrier rate in the country. The present article reviews the literature on the treatment response of HBV prevalent in Pakistani population. The average treatment response of Lamivudine and interferon-α is 25.81% and 47.95%, respectively. Peg-Interferon was shown to be not effective against the HBV/HCV (hepatitis C virus)/HDV (hepatitis Delta virus) co-infection. The present study reveals that interferon-α is the most effective therapy available for HBV infection prevalent in Pakistani population. Genotype C & D are the most common HBV genotypes in Pakistan and are associated with increased severity and less response to interferon therapy. This poses a great challenge for physicians and researchers and further studies are needed to describe the outcome of the current therapies recommended against HBV infection in Pakistani population.

Hepatitis C treatment: current and future perspectives.

Hepatitis C virus (HCV) is a member of Flaviviridae family and one of the major causes of liver disease. There are about 175 million HCV infected patients worldwide that constitute 3% of world's population. The main route of HCV transmission is parental however 90% intravenous drug users are at highest risk. Standard interferon and ribavirin remained a gold standard of chronic HCV treatment having 38-43% sustained virological response rates. Currently the standard therapy for HCV is pegylated interferon (PEG-INF) with ribavirin. This therapy achieves 50% sustained virological response (SVR) for genotype 1 and 80% for genotype 2 & 3. As pegylated interferon is expensive, standard interferon is still the main therapy for HCV treatment in under developed countries. On the other hand, studies showed that pegylated IFN and RBV therapy has severe side effects like hematological complications. Herbal medicines (laccase, proanthocyandin, Rhodiola kirilowii) are also being in use as a natural and alternative way for treatment of HCV but there is not a single significant report documented yet. Best SVR indicators are genotype 3 and 2, < 0.2 million IU/mL pretreatment viral load, rapid virological response (RVR) rate and age <40 years. New therapeutic approaches are under study like interferon related systems, modified forms of ribavirin, internal ribosome entry site (HCV IRES) inhibitors, NS3 and NS5a inhibitors, novel immunomodulators and specifically targeted anti-viral therapy for hepatitis C compounds. More remedial therapies include caspase inhibitors, anti-fibrotic agents, antibody treatment and vaccines.

pH-responsive high stability polymeric nanoparticles for targeted delivery of anticancer therapeutics.

The practical application of nanoparticles (NPs) as chemotherapeutic drug delivery systems is often hampered by issues such as poor circulation stability and targeting inefficiency. Here, we have utilized a simple approach to prepare biocompatible and biodegradable pH-responsive hybrid NPs that overcome these issues. The NPs consist of a drug-loaded polylactic-co-glycolic acid (PLGA) core covalently 'wrapped' with a crosslinked bovine serum albumin (BSA) shell designed to minimize interactions with serum proteins and macrophages that inhibit target recognition. The shell is functionalized with the acidity-triggered rational membrane (ATRAM) peptide to facilitate internalization specifically into cancer cells within the acidic tumor microenvironment. Following uptake, the unique intracellular conditions of cancer cells degrade the NPs, thereby releasing the chemotherapeutic cargo. The drug-loaded NPs showed potent anticancer activity in vitro and in vivo while exhibiting no toxicity to healthy tissue. Our results demonstrate that the ATRAM-BSA-PLGA NPs are a promising targeted cancer drug delivery platform.

Hearing improvement after stapedotomy using teflon loop prosthesis.

OBJECTIVE: To assess hearing improvement after stapedotomy for otosclerosis using teflon loop prosthesis. DESIGN: Quasy experimental study. PLACE AND DURATION OF STUDY: January 2001 to May 2003 in the Otorhinolaryngology Department, CMH, Rawalpindi. PATIENTS AND METHODS: Thirty diagnosed cases of otosclerosis were included in the study. Ear with greater air-bone gap was selected and stapedotomy was done using teflon loop prosthesis (size 4 - 4.5 mm). Mild vertigo occurred during immediate postoperative period, which subsided with Inj. Prochlorperazine. Patients were followed-up postsurgically at one month, two months, four months, six months and one year and postoperative air-bone gap was calculated. RESULTS: Out of 30 cases, there were 24 males and 06 females. The age ranged from 18 to 50 years. Twenty-one (70%) patients had bilateral hearing loss and 09 (30%) had tinnitis also. Pre-operative audiograms showed conductive deafness. Carhart's notch was present in 10 (33.3%) cases. Tympanogram revealed loss of stapedial reflex. Postoperatively good hearing improvement was seen in 56.7% (postoperative air-bone gap closure upto 10 dB), fair improvement in 30% (postoperative air bone gap closure from 11 to 20 dB) and poor results in 10% (postoperative air-bone gap more than 21 dB) cases. One patient developed dead ear. CONCLUSION: Stapedotomy using teflon loop prosthesis for otosclerosis is an effective method to restore hearing. Complications are uncommon.

Controlled delivery of valsartan by cross-linked polymeric matrices: Synthesis, in vitro and in vivo evaluation.

The purpose of study was to develop chemically cross-linked chitosan-co-poly(AMPS) hydrogel based on low molecular weight chitosan for pH-responsive and controlled drug delivery of a model drug. Cross-linking was achieved chemically, by using free radical polymerization technique. Polymer (low molecular weight chitosan) was chemically cross-linked with monomer (2-acrylamido-2-methylpropane sulfonic acid) in aqueous medium. N, N'-Methylenebisacrylamide (MBA) was used as cross-linking agent. Sodium hydrogen sulfite (SHS) and ammonium peroxodisulphate (APS) were used as initiators in a chemical reaction. Hydrogels were characterized by FT-IR, SEM and DSC. Swelling studies and pH-sensitivity of hydrogels were studies at pH 1.2 and 7.4. Chitosan-co-poly(AMPS) hydrogels were administered to rabbits orally to evaluate its pharmacokinetic behavior. As a result of successful cross-linking of polymer and monomer, novel co-polymer has been developed, having suitable characteristics as desired for controlled release drug delivery system. Maximum swelling, drug loading and release have been observed at pH 7.4. In vivo results exhibited significant drug release and absorption at pH 7.4 in rabbits. It is concluded that highly swelling chitosan-AMPS based hydrogels were developed having pH independent swelling and pH dependent drug release properties. These hydrogels have great potential to be used for loading and controlled release of various therapeutic agents.