Synthesis and in Silico Docking Studies of Ethyl 2-(2-Arylidene-1-alkylhydrazinyl)thiazole-4-carboxylates as Antiglycating Agents.

Ethyl 2-(2-arylidene-1-alkylhydrazinyl)thiazole-4-carboxylates (1a-k) were synthesized by alkylation on HN- of ethyl 2-(2-arylidenehydrazinyl)thiazole-4-carboxylates. The proposed structures (1a-k) are corroborated by spectro-analytical techniques like UV, FT-IR, 1 H-, 13 C-NMR and HR-MS. All synthesized compounds were screened for their antiglycation and antioxidant assays. The in vitro antiglycation results revealed promising activity of compounds 1a, 1b, 1d, 1e, 1f, 1g, 1j and 1k with IC50 values 0.0004±1.097-17.22±0.538 µM when compared to standard, aminoguanidine (IC50 =25.50±0.337 µM). Among all tested compounds 1j and 1k are the best antiglycating agents with IC50 values 1.848±0.646 and 0.0004±1.097 µM, respectively. The in-silico studies also agree with these results where binding energy for 1j and 1k was found to be -9.25 and -8.42 kcal/mol with calculated dissociation constants of 0.16 and 0.67 µM, respectively. The antiglycation results demonstrate the application of these compounds in reducing diabetic complications.

Sequence variants in four genes underlying Bardet-Biedl syndrome in consanguineous families.

PURPOSE: To investigate the molecular basis of Bardet-Biedl syndrome (BBS) in five consanguineous families of Pakistani origin. METHODS: Linkage in two families (A and B) was established to BBS7 on chromosome 4q27, in family C to BBS8 on chromosome 14q32.1, and in family D to BBS10 on chromosome 12q21.2. Family E was investigated directly with exome sequence analysis. RESULTS: Sanger sequencing revealed two novel mutations and three previously reported mutations in the BBS genes. These mutations include two deletions (c.580_582delGCA, c.1592_1597delTTCCAG) in the BBS7 gene, a missense mutation (p.Gln449His) in the BBS8 gene, a frameshift mutation (c.271_272insT) in the BBS10 gene, and a nonsense mutation (p.Ser40*) in the MKKS (BBS6) gene. CONCLUSIONS: Two novel mutations and three previously reported variants, identified in the present study, further extend the body of evidence implicating BBS6, BBS7, BBS8, and BBS10 in causing BBS.

Biological activities of Allium sativum and Zingiber officinale extracts on clinically important bacterial pathogens, their phytochemical and FT-IR spectroscopic analysis.

The spread of bacterial infectious diseases is a major public threat. Herbs and spices have offered an excellent, important and useful source of antimicrobial agents against many pathological infections. In the current study, the antimicrobial potency of fresh, naturally and commercial dried Allium sativum and Zingiber officinale extracts had been investigated against seven local clinical bacterial isolates such as Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Staphylococcus aureus, Streptococcus pyogenes, Staphylococcus epidermidis, and Serratia marcesnces by the agar disc diffusion method. All tested pathogens except P. aeruginosa and E. coli were most susceptible to ethanolic and methanolic extracts of A. sativum. Similarly, chloroform and diethyl ether extracts of Z. officinale showed a greater zone of inhibition of tested pathogens except for P. aeruginosa and E. coli. We found that all extracts of A. sativum and Z. officinale have a strong antibacterial effect compared to recommended standard antibiotics through activity index. All results were evaluated statistically and a significant difference was recorded at P< 0.05. Antioxidant activity of extracts showed that 10 out of 13 extracts have high scavenging potential. Thin layer chromatography profiling of all extracts of A. sativum and Z. officinale proposed the presence of various phytochemicals such as tannins, phenols, alkaloids, steroids and saponins. Retention factor of diverse phytochemicals provides a valuable clue regarding their polarity and the selection of solvents for separation of phytochemicals. Significant inhibition of S. aureus was also observed through TLC-Bioautography. FT-IR Spectrometry was also performed to characterize both natural and commercial extracts of A. sativum and Z. officinale to evaluate bioactive compounds. These findings provide new insights to use A. sativum and Z. officinale as potential plant sources for controlling pathogenic bacteria and potentially considered as cost-effective in the management of diseases and to the threat of drug resistance phenomenon.

Comparative study of antioxidant, metal chelating and antiglycation activities of Momordica charantia flesh and pulp fractions.

Momordica charantia is commonly used as a vegetable and folk medicine in most parts of South Asia. This study aims to determine and compare the antioxidant, metal chelating and antiglycation activities of aqueous extracts of M. charantia fruit flesh (MCF) and fruit pulp (MCP) fractions. Our results show that MCP has pronounced DPPH and ABTS radical scavenging potential compared to MCF. In the antiglycation assay both fractions illustrated considerable inhibitory activities against the formation of AGEs induced by glucose with an efficacy of 75 and 67% with 150 µl of MCP and MCF extracts respectively, almost equal to 0.3mM amino guanidine. Results for metal catalysed protein fragmentation and autoxidative and glycoxidation assays demonstrate that MCF and MCP inhibited metal catalysed protein fragmentation. The percentage of relative standard deviation for three replicate measurements of 150 µl of MCF and MCP was < 3.0% for antiglycation. The antioxidant assays with regression values of MCP (0.981 and 0.991) and MCF (0.967 and 0.999) were also recorded. We conclude that both extracts possess high antioxidant and antiglycation activities and are equally good sources of antioxidant and antiglycating agents.

Short communication: in vitro assessment of antioxidant, antibacterial and phytochemical analysis of peel of Citrus sinensis.

Antibacterial effect of Citrus sinensis peel extracts was evaluated against several pathogenic bacteria associated with human and fish infections viz., Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumonia, Staphylococcus aureus, Streptococcus pyogenes, Staphylococcus epidermidis, Serratia marcesnces, Shigella flexneri, Enterobacter amnigenus, Salmonella Typhimurium and Serratia odorifera. Methanol, ethanol, chloroform and diethyl ether solvents were used for extraction. In vitro antibacterial activity was analyzed by agar well and agar disc diffusion methods. It was found that ethanol extract showed highly significant inhibition of E. coli and K. pneumonia (12.6±0.94 mm and 11.6±1.2 mm) whereas methanol extract of C. sinensis also showed high zone of inhibition of S. odorifera (10.0±2.16 mm). The potential activity of active extracts was assessed and also compared with standard antibiotics through activity index formulation. The order of antioxidant activity through ABTS·+ and DPPH free radical scavenging activity was ethanol>methanol>chloroform>diethyl ether. Phytochemical screening of all solvents had determined the presence of terpenoids, alkaloids, steroids, glycosides and flavonoids. It was also found that Chloroform/Methanol (5:5) and Butanol/Ethanol/Water (4:1:2.2) solvent systems showed significant separation of active phytochemical constituents. These findings reveal the potential use of C. sinensis peel to treat infectious diseases, which are being caused by microorganisms.

EDTA functionalized pine needle biochar (EDTA@BC); a valorized bio-material for removal of Ni(II) from aqueous solution.

The preparation of ethylenediaminetetraacetic acid (EDTA) functionalized pine needles biochar (EDTA@BC) as a low-cost active adsorbent and its effectiveness in removing Ni(II) from aqueous solution at various conditions is reported in this paper. First, alkali activation was selected to render the pine needle biochar with an excellent porous structure and increased concentration of hydroxyl groups to facilitate grafting. Subsequently, a simple method was utilized to graft EDTA onto the biochar. The prepared EDTA@BC was characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy dispersive x-ray spectrometry (EDX). Batch adsorption studies were conducted to assess the impact of various parameters such as solution pH, adsorbent dosage, adsorbate volume, and shaking time on the removal efficiency of Ni(II). At pH 6, 100 mg dosage, 4 mL of adsorbate volume, and 10 min of shaking time, the maximum removal efficiency of Ni(II) was observed to be 89%. EDTA@BC showed reasonable sorption performance still after the third cycle of regeneration. The effect of interfering ions such as Pb, Cr, Cu, and Hg was evaluated, resulting a decrease of 69%, 78%, 76%, and 68%, respectively, in its sorption capacity. The Langmuir model provided a better fit for Ni(II) in the concentration range of 0.1-2000 ppm under optimized conditions, with qmax of 46.69 ± 1.031 mg/g and KL of 0.001, compared with the Freundlich isotherm, which yielded n = 0.234 and χ2 = 2.7899, Temkin isotherm (R2 = 0.9520), and Redlich-Peterson isotherm (R2 = 0.9725). The removal of Ni(II) by EDTA@BC was found to be the pseudo-second-order kinetics. Thermodynamic studies indicated adsorption process to be endothermic and nonspontaneous. Hence, a sustainable valorized bio-material (EDTA@BC) is prepared having better sorption efficiency of Ni(II) from aqueous solution with possible wide applicability. RESEARCH HIGHLIGHTS: New EDTA functionalized indigenous pine needles biochar (EDTA@BC) was prepared. This low-cost active adsorbent found effective in removing Ni(II) from aqueous solution. FTIR, SEM, and EDX proved synthesis and uptake of Ni(II) from aqueous solution. Ni(II) removal, regeneration, interfering and adsorption studies were performed by UV-Vis spectroscopy.

Retracted: Partial purification and characterization of lipase from locally produced edible oil-seeds and its relevance in industries.

Lipase was extracted from germinating seeds of Helianthus annus (Sunflower), Zea mays (Maize), and Brassica compastris (Mustard). The lipolytic activity was assessed using olive oil as substrate at different germination-time and the maximum-activity was obtained after 120 hr. Partial-purification was executed by precipitating the seed-homogenate with varying concentration of ammonium sulfate solution. 80% ammonium sulfate solution showed maximum lipase activity of 5320IUml-1 , 3500IUml-1 , 3080IUml-1 with 9.6, 6.9, and 4.8-fold purification and total protein content of 162, 84, and 60 mg for partially purified enzyme extracts namely SN5, BN5, and MN5, respectively. The optimum temperature and pH observed for hydrolysis of olive oil were 37°C, and 8.0 respectively. Enzyme was found to be stable upto 6 days at 4°C and its activity was stimulated by Ca+2 ions. Oil-stains removal from cotton fabric was observed to be superior in the presence of lipase and detergent. Moreover, the SN5, BN5, and MN5 lipase increased free fatty acid release upto 4.2, 4.3, and 3.8 mg, respectively than wastewater without treatment of lipase (0.21 mg) and promoted fat hydrolysis to approximately 40, 42, and 48% mass reduction after 6 hr incubation of fat particle at a concentration of 20 mg/ml. Biodiesel produced by catalyzing transesterification of vegetable oil with SN5, BN5, and MN5 lipase provided an acid value of 0.8, 1.08, and 0.5 mg/g, viscosity 5.50, 5.7, and 5.53 mm2 /s and density 0.87, 0.88, and 0.79 g/ml, respectively. To the best of our knowledge, no such study has been conducted prior on lipase from the seeds mentioned above in Azad Kashmir region.

Novel heterozygous sequence variant in the GLI1 underlies postaxial polydactyly.

Polydactyly is one of the most common congenital abnormal phenotype of autopod, which is characterized by extra supernumerary digit in hands/feet with or without well-developed bony structure within the digits. Preaxial polydactyly (PPD), postaxial polydactyly (PAP), and meso-axial (central) polydactyly are three different isoforms of polydactyly. Genetically, at least 10 genes have been identified causing nonsyndromic polydactyly. In the present study, we have investigated a large family segregating autosomal dominant form of nonsyndromic polydactyly. Whole exome sequencing followed by Sanger sequencing revealed a novel heterozygous missense variant (NM_005269.3; c.1064C>A; p.(Thr355Asn) in the gene GLI1 segregating with the disease phenotype within the family. This study presents first familial case of autosomal dominant form of polydactyly caused by the GLI1 variant.

Chromium removal from aqueous solutions using new silica gel conjugates of desferrioxamine or diethylenetriaminepentaacetic acid.

Desferrioxamine (DFO) and diethylenetriaminepentaacetic acid (DTPA) conjugated with silica gel (IDFOSG and IDTPASG, respectively) were evaluated as adsorbents for chromium in aqueous solutions. Different parameters affecting adsorption such as pH, sorbent dosage, contact time, sample volume and potential of interfering ions have been optimized. The optimum pH for chromium binding was 4 for 100 mg of adsorbents at 5 min of table shaking with 5 mL sample volume of chromium solutions. Langmuir adsorption model described the removal of chromium ions. The adsorption capacity for chromium was 90% for IDFOSG and 83% for IDTPASG in single solutions, and at least 75% in multielemental solutions. Considering the removal efficacy, regeneration and stability, DFO-grafted silica gel was generally superior to its DTPA counterpart and may be applied to the removal of traces of chromium species from natural waters.

Variants in GLI3 Cause Greig Cephalopolysyndactyly Syndrome.

Background: Greig cephalopolysyndactyly syndrome (GCPS) is a disorder of autopod and craniofacial abnormalities. Autopod anomalies include preaxial and/or postaxial polydactyly together with or without syndactyly while craniofacial features include hypertelorism and macrocephaly. GCPS is inherited in an autosomal dominant manner and is caused by sequence variants in GLI3. Methodology and Results: In this study, we examined four unrelated families with GCPS segregating in an autosomal dominant manner. Sanger sequencing revealed three novel (p.Tyr146Leufs*19, p.Glu99Serfs*60, and p.Thr541Arg) and one previously reported non-sense variant (p.Arg792*) in GLI3. Conclusion: The study expands the spectrum of the variants in the GLI3 gene linked to GCPS, and should also facilitate genetic counseling of GCPS patients in the Pakistani population.

Natural products as anti-glycation agents: possible therapeutic potential for diabetic complications.

Diabetes mellitus is characterised by hyperglycaemia, lipidaemia and oxidative stress and predisposes affected individuals to long-term complications afflicting the eyes, skin, kidneys, nerves and blood vessels. Increased protein glycation and the subsequent build-up of tissue advanced glycation endproducts (AGEs) contribute towards the pathogenesis of diabetic complications. Protein glycation is accompanied by generation of free radicals through autoxidation of glucose and glycated proteins and via interaction of AGEs with their cell surface receptors (referred to as RAGE). Glycationderived free radicals can damage proteins, lipids and nucleic acids and contribute towards oxidative stress in diabetes. There is interest in compounds with anti-glycation activity as they may offer therapeutic potential in delaying or preventing the onset of diabetic complications. Although many different compounds are under study, only a few have successfully entered clinical trials but none have yet been approved for clinical use. Whilst the search for new synthetic inhibitors of glycation continues, little attention has been paid to anti-glycation compounds from natural sources. In the last few decades the traditional system of medicine has become a topic of global interest. Various studies have indicated that dietary supplementation with combined anti-glycation and antioxidant nutrients may be a safe and simple complement to traditional therapies targeting diabetic complications. Data for forty two plants/constituents studied for anti-glycation activity is presented in this review and some commonly used medicinal plants that possess anti-glycation activity are discussed in detail including their active ingredients, mechanism of action and therapeutic potential.

Pakistamide C, a new sphingolipid from Abutilon pakistanicum

The Abutilon genus from the Malvaceae family is of medicinal importance, and members of this genus are distributed in the tropical and subtropical regions of Asia and other parts of the world. Abutilon pakistanicum Jafri & Ali is mainly found in Pakistan. It has been used by different systems of traditional medicines to treat different diseases. Pakistamide C, a new sphingolipid, has been isolated from the ethyl acetate soluble fraction of the methanolic extract of A. pakistanicum. Different spectroscopic techniques such as NMR (1H, 13C, COSY, NOESY, HSQC, HMBC) and Mass spectrometry (EI-MS, and FAB-MS experiments) were used to elucidate the structure of pakistamide C.