Characterization of indigenous Pyrus germplasm of Azad Jammu and Kashmir revealed by SDS-PAGE analysis

Soluble proteins of sixty pear genotypes/varieties were extracted from their leaves, separated by polyacrylamide gel electrophoresis and distinguished by protein banding pattern. Seven types of protein with 12 to 20 bands were observed. SD45 (Kashmiri nakh) showed the maximum (20) and KT60 (Keiffer) the minimum (12) number of bands. Remaining accessions exhibited less variability having 15 to 19 bands. The accessions were classified into 12 groups and individual accessions varied from 0.71to 0.97 similarity level. Moreover, the highest similarity was expressed among groups 1 to 7. UPGMA cluster analysis distributed the accessions into three clusters, seven sub-clusters along with 11 identical groups, one independent group and two independent accessions. There were 37 accessions in cluster I, 16 in II, 3 in III, 2 in independent group 12 and 2 accessions existed independently. The most variable accession, KT60 (Keiffer) fell independently, had the highest genetic diversity. The findings show that the pear accessions have different protein profile irrespective of their geographic locations and climatic conditions

Determination of genetic variability of Asian rice (Oryza sativa L.) varieties using microsatellite markers

The microsatellite or simple sequence repeat (SSR) marker analysis was done to determine the allelic diversity and relationship among thirty-five Asian cultivars of rice including 19 aromatic, 13 nonaromaticand 3 japonica type cultivars. A total of 144 alleles were detected at the 32 SSR loci, of which 141 (98%) were polymorphic. The number of alleles generated by each marker ranged from 2 to 13 with an average of 4.5 alleles per marker. The size of smallest and largest allele ranged from 8(RM122) to as high as 71 (RM302). Polymorphism information content (PIC) values ranged between 0.157 (RM19, RM55) and 0.897 (RM70), with an average of 0.603 per marker. Basmati rice varieties amplified different alleles at 15 of the SSR loci than those in the japonica and/ or indica rice varieties. A number of SSRs were identified that could be utilized to differentiate between basmati and other non-basmati rice varieties. The RM252 and RM310 showed a clear differentiation of japonica cultivarsfrom other ones. Pair-wise Nei and Li’s similarity coefficients ranged from 0.19 to 0.90. The dendrogram based on the cluster analysis by microsatellite polymorphism, grouped 35 rice cultivars into two major groups effectively differentiating the tall, late maturing and slender aromatic cultivars from the short statured, early, short bold and long bold non-aromatic cultivars. These results could be useful for monitoring purity, genotype identification and for plant variety protection

Effect of Permeation Enhancers on the Release Behavior and Permeation Kinetics of Novel Tramadol Lotions

Purpose: The aim of this research work was to formulate, characterize and evaluate the in vitro permeation behavior of tramadol lotion containing propylene glycol (PG) and polyethylene glycol (PEG) as permeation enhancers.Methods: The permeation experiments were conducted in vitro using full thickness rabbit skin in Franz diffusion cells. The donor compartment was filled with PBS (phosphate buffered saline) at pH 7.4 ± 0.1. The receptor phase was continuously stirred PBS (pH 7.4) at 37 °C ± 0.5. The amount of tramadol permeated into the receptor phase was determined spectrophotometrically at 271 nm. Various permeation parameters such as permeation coefficient (Kp), diffusion coefficient (D), flux (J), input rate,and enhancement ratio were obtained using Fick’s diffusion laws.Results: Permeation increased with increase in the concentrations of both enhancers tested. Maximum cumulative amount permeated for control lotion (Lc) was 357 ìg/cm2/min with input rate 0.574 ìg/min and lag time (tlag) of 34.93 min, while for the optimum test lotion (L4, containing 8 % PG/PEG in ratio of 1:1 v/v), it was 926 ìg/cm2/min, 1.482 ìg/min and 58.36 min, respectively. The significantly (p < 0.05) higher permeability shown by the test lotion L4 can be attributed, in part, to the interaction of PG withintercellular lipids leading to the disruption of their organization and increasing their fluidity, and also partly as a result of solubilization of lipid bilayers by PEG.Conclusion: A binary system of PG and PEG in lotion can be successfully utilized for the permeation enhancement of tramadol.Keyword: Tramadol, Transdermal delivery, Permeation, Propylene glycol, Polyethylene glycol, Rabbit skin

Hypoglycemic effect of white (Morus alba L.) and black (Morus nigra L.) mulberry fruits in diabetic rat

The aim of the present study was to investigate the hypoglycemic effect of white (Morus alba L.) and black (Morus nigra L.) mulberry fruits either used individually or in a combination on alloxan diabetic rats. These fruits are reported to be rich in antioxidants, flavonoids and phenolics that can potentially fight against diabetes mellitus. Male albino rats were divided into 5 groups: normal control, alloxan-diabetic control, diabetic rats treated with white mulberry fruit powder at 5% in the diet, diabetic rats treated with black mulberry fruit powder at 5% in the diet and diabetic rats treated with mixture of white and black mulberry fruits powder at 5% in the diet. After 4 weeks of treatment, blood glucose level, liver and kidney enzymes activity, lipid profile, lipid peroxidation and histopathological studies on liver, kidney and pancreas were evaluated. The mixture of white and black mulberry fruits showed the most significant (p < 0.05) improvement in feed efficiency ratio with increasing body weight gain, as well as decrease in blood glucose level and liver-kidney dysfunction when compared with diabetic control rats. Significant decrease in serum cholesterol, triglycerides and low density lipoprotein cholesterol (LDLc) as well as significant increase in high density lipoprotein cholesterol (HDLc) in diabetic rats was observed with all treatments. Moreover, mulberry fruits administration caused significant inhibition in lipid peroxidation and α-amylase activity. In addition, the beneficial effect of all treatments was further confirmed with histopathological examination of liver, kidney and pancreas. This study reveals hypoglycemic and hypolipidemic effects of white and black mulberry fruits either used individually or in combination as a dietary supplement in alloxan diabetic rats

Synthesis of thermochemically stable tetraphenyladamantane-based microporous polymers as gas storage materials

In view of environmental pollution control and purification of natural gases, developing ideal porous materials for small gas molecule (hydrogen, methane and carbon dioxide) capture is an important, pressing challenge. Accordingly, herein, three microporous organic polymers (MOP-Ad) have been synthesized by Suzuki coupling polymerization of 1,3,5,7-tetrakis(4-bromophenyl)adamantane “knots” with three phenylboronic acid-type “rods”. Gas adsorption studies of the MOP-Ad materials demonstrated their permanent porosity and good gas storage capabilities (1.07 wt% H2 at 77.3 K and 1.13 bar, 10.3 wt% CO2 and 2.4 wt% CH4 at 273.1 K and 1.13 bar), as well as moderate CO2/CH4 adsorption selectivity. Moreover, high thermal stability (up to 520 °C) and remarkable chemical resistance to strong acids and bases were found in these polymers, making them suitable candidates as gas storage materials in harsh chemical environments

Glycation marker glucosepane increases with the progression of osteoarthritis and correlates with morphological and functional changes of cartilage in vivo

Background: Changes of serum concentrations of glycated, oxidized, and nitrated amino acids and hydroxyproline and anticyclic citrullinated peptide antibody status combined by machine learning techniques in algorithms have recently been found to provide improved diagnosis and typing of early-stage arthritis of the knee, including osteoarthritis (OA), in patients. The association of glycated, oxidized, and nitrated amino acids released from the joint with development and progression of knee OA is unknown. We studied this in an OA animal model as well as interleukin-1β-activated human chondrocytes in vitro and translated key findings to patients with OA. Methods: Sixty male 3-week-old Dunkin-Hartley guinea pigs were studied. Separate groups of 12 animals were killed at age 4, 12, 20, 28 and 36 weeks, and histological severity of knee OA was evaluated, and cartilage rheological properties were assessed. Human chondrocytes cultured in multilayers were treated for 10 days with interleukin-1β. Human patients with early and advanced OA and healthy controls were recruited, blood samples were collected, and serum or plasma was prepared. Serum, plasma, and culture medium were analyzed for glycated, oxidized, and nitrated amino acids. Results: Severity of OA increased progressively in guinea pigs with age. Glycated, oxidized, and nitrated amino acids were increased markedly at week 36, with glucosepane and dityrosine increasing progressively from weeks 20 and 28, respectively. Glucosepane correlated positively with OA histological severity (r = 0.58, p < 0.0001) and instantaneous modulus (r = 0.52–0.56; p < 0.0001), oxidation free adducts correlated positively with OA severity (p < 0.0009–0.0062), and hydroxyproline correlated positively with cartilage thickness (p < 0.0003–0.003). Interleukin-1β increased the release of glycated and nitrated amino acids from chondrocytes in vitro. In clinical translation, plasma glucosepane was increased 38% in early-stage OA (p < 0.05) and sixfold in patients with advanced OA (p < 0.001) compared with healthy controls. Conclusions: These studies further advance the prospective role of glycated, oxidized, and nitrated amino acids as serum biomarkers in diagnostic algorithms for early-stage detection of OA and other arthritic disease. Plasma glucosepane, reported here for the first time to our knowledge, may improve early-stage diagnosis and progression of clinical OA

Analysis of serum advanced glycation endproducts reveals methylglyoxal-derived advanced glycation MG-H1 free adduct is a risk marker in non-diabetic and diabetic chronic kidney disease

Accumulation of advanced glycation endproducts (AGEs) is linked decline in renal function, particularly in patients with diabetes. Major forms of AGEs in serum are protein-bound AGEs and AGE free adducts. In this study, we assessed levels of AGEs in subjects with and without diabetes, with normal renal function and stages 2 to 4 chronic kidney disease (CKD), to identify which AGE has the greatest progressive change with decline in renal function and change in diabetes. We performed a cross-sectional study of patients with stages 2 – 4 CKD, with and without diabetes, and healthy controls (n = 135). Nine protein-bound and free adduct AGEs were quantified in serum. Most protein-bound AGEs increased moderately through stages 2 – 4 CKD whereas AGE free adducts increased markedly. Methylglyoxal-derived hydroimidazolone MG-H1 free adduct was the AGE most responsive to CKD status, increasing by 8-fold and 30-fold in stage 4 CKD in patients without and with diabetes, respectively. MG-H1 Glomerular filtration flux was increased 5-fold in diabetes, likely reflecting increased methylglyoxal glycation status. We conclude that serum MG-H1 free adduct concentration was strongly related to stage and increased in diabetes status. Serum MG-H1 free adduct is a candidate AGE risk marker of non-diabetic and diabetic CKD

Glucose metabolite glyoxal induces senescence in telomerase-immortalized human mesenchymal stem cells

Abstract Background Various by-products of the cellular metabolism, such as reactive carbonyl species (RCS) are potentially harmful to cells and tissues, and play a role in many physiological and pathological processes. Among various RCS is the highly reactive dicarbonyl glyoxal (GO), which is a natural physiological metabolite produced by the auto-oxidation of glucose, and can form covalent adducts known as advanced glycation endproducts (AGE). We have previously reported that GO accelerates ageing and causes premature senescence in normal human skin fibroblasts. Results Using a bone marrow-derived telomerase-immortalised mesenchymal stem cell line hMSC-TERT we have observed that an exposure of cells to 0.75 mM and 1 mM GO induces irreversible cellular senescence within 3 days. Induction of senescence in hMSC-TERT was demonstrated by a variety of markers, including characteristic cell morphology and enlargement, vacuolisation, multinucleation, induction of senescence associated β-galactosidase, cell cycle arrest, and increased levels of a cell cycle inhibitor p16. These changes were accompanied by increased extent of DNA breaks as measured by the comet assay, and increased levels of the AGE product, carboxymethyl-lysine (CML). Furthermore, the in vitro differentiation potential of hMSC-TERT to become functional osteoblasts was highly reduced in GO-treated stem cells, as determined by alkaline phosphatase (ALP) activity and mineralized matrix (MM) formation. Conclusions The results of our study imply that an imbalanced glucose metabolism can reduce the functioning ability of stem cells in vivo both during ageing and during stem cell-based therapeutic interventions.</p

New insights on the complex dynamics of two-phase flow in porous media under intermediate-wet conditions

Multiphase flow in porous media is important in a number of environmental and industrial applications such as soil remediation, CO2 sequestration, and enhanced oil recovery. Wetting properties control flow of immiscible fluids in porous media and fluids distribution in the pore space. In contrast to the strong and weak wet conditions, pore-scale physics of immiscible displacement under intermediate-wet conditions is less understood. This study reports the results of a series of two-dimensional high-resolution direct numerical simulations with the aim of understanding the pore-scale dynamics of two-phase immiscible fluid flow under intermediate-wet conditions. Our results show that for intermediate-wet porous media, pore geometry has a strong influence on interface dynamics, leading to co-existence of concave and convex interfaces. Intermediate wettability leads to various interfacial movements which are not identified under imbibition or drainage conditions. These pore-scale events significantly influence macro-scale flow behaviour causing the counter-intuitive decline in recovery of the defending fluid from weak imbibition to intermediate-wet conditions