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Seaweeds have been used as nutraceuticals because of certain metabolites present in some species including polyphenols. Seven species of seaweeds collected from the Karachi coast were screened for the first time both for phycochemical analysis and pharmacological activities. The phycochemicals quantified included phenols, flavonoids, tannins and saponins. Melanothamnus afaqhusainii showed the highest content of phenols (2.16mg GAE/g) while highest flavonoids were observed in Coelarthrum muelleri (4.59mg RE/g). Tannins were found in low amounts while saponins were observed as major constituents among the seaweeds ranging from 1.34-3.47% in Sargassum swartzii and Codium flabellatum, respectively. Saponins were not analysed in Rhodophyta due to gel formation. Pharmacological screening revealed analgesic and anti-inflammatory effects. Both the methods of analgesic activities have shown significant increase in reaction time when methanol extracts were used. The reason for delayed anti-inflammatory activity of Solieria robusta and C. muelleri was found correlating with its gel forming ability. While Cystoseira indica and C. flabellatum exhibited highly significant effect from 1st to 5th h. Results suggested that selected seaweeds had potential in combating both acute and chronic inflammation and pain and hence can be used for therapeutic efficacy. [ABSTRACT FROM AUTHOR]

Phycochemical and pharmacological studies were carried out on Ulva fasciata Delile. The ash content was found as 20.4812 % dry weight, moisture content 14.5514 %, total fat content as 0.1878% and 0.49341 %. Total carbohydrate was found as 54.5301-54.2246% dry weight, phenolic content as 0.022%, flavonoids found to be 0.0313% and tannins were 0.00003 %. Ulva fasicata showed central analgesic activity and significant anti-inflammatory activity at the dose of 400 mg/kg bw. [ABSTRACT FROM AUTHOR]

A comprehensive phytochemical analyses of six different botanicals obtained from Sargassum wightii on GC-MS has revealed the presence of various chemicals. All these identifications were further authenticated by the retention indices and relative retention times. These extracts and fractions contained volatile, non-polar and moderately polar constituents among which 81 were identified. Out of these, 33 were natural constituents while 48 were anthropogenic contaminants. Identified phytochemicals belong to different classes of natural products including 23 fatty acids, 2 fatty aldehydes, 2 fatty alcohols, 2 phytosterols and 4 terpenoids. 15 phytochemicals including 5 fatty acids, methyl iso-pentadecanoate (37), methyl pentadecylate (40), isopropyl myristate (41), methyl elaidate (56) and 9Z,12EOctadecadienoic acid ME (60); 2 fatty aldehydes, nonanal (16) and 2E-decenal (20); 2 fatty alcohols, lignocerol (68) and pentacosyl alcohol (73); 4 isoprenoids, phytane (36), iso-phytol (42), hexahydrofarnesyl acetone (43) and E-phytol (57) and 2 phytosteroids, -Sitosterol (79) and fucosterol (81) were identified for the first time from S. wightii. 48 environmental contaminants included 28 petrochemicals (17 aliphatic, 6 aromatic and 5 oxygenated hydrocarbons), 14 cyclosiloxanes and 6 phthalates. Few of these hydrocarbons are reported from different seaweeds including S. wightii but current study is reporting these as environmental contaminant. Similarly identified hydroperoxides 1-Methyl-pentyl hydroperoxide (9) and 1-ethyl-butyl hydoperoxide (10) could be the oxidative products of 2- hexanone (3) and 3-hexanone (4), respectively and could be the biotransformed products of the assimilated ketones 3 and 4. Overall current study is confirming identification of 15 constituents for the first time from S. wightii. Antioxidant activities of these extracts and fractions were performed using DPPH, ABTS, FRAP, TAC protocols. Botanicals obtained using n-hexane and methanol has shown remarkable results comparable with commercial antioxidants, BHT and BHA. [ABSTRACT FROM AUTHOR]

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Up to now, the direct ligation of two DNA fragments with opposite directions to obtain 3'-3' or 5'-5' phosphate ester bonds is still challenging. The only way to obtain DNA oligonucleotides containing a 3'-3' or 5'-5' inversion of polarity sites is based on professionalDNAchemical synthesis. Herein, we demonstrate a convenient template-directed chemical ligation that enables 3'-3' and 5'-5' linkages of two DNA oligonucleotides. This method is based on the assembly of two oligonucleotides on a template in opposite directions through forming antiparallel and parallel duplexes simultaneously, followed by coupling with N-Cyanoimidazole under mild condition. Moreover, on the basis of DNA oligonucleotides with 5'-5' linkage obtained through our template-directed chemical ligation, we developed a new cDNA display technique for in vitro selection of functional polypeptides. [ABSTRACT FROM AUTHOR]

Purpose: The purpose of this paper is to map the locus for a variant form of Oguchi disease in a Pakistani family and to identify the causative mutation., Methods: Family 61029 was ascertained in the Punjab province of Pakistan. It includes three 13- to 19-year-old patients with night blindness and 12 unaffected family members. A complete ophthalmological examination including fundus photography and electroretinography (ERG) was performed on each family member. A genome-wide scan was performed using microsatellite markers at about 10 cM intervals, and two-point lod scores were calculated. Polymerase chain reaction (PCR) cycle dideoxynucleotide sequencing was used to screen candidate genes inside the linked region for mutations and to delineate the deletion. Multiplex PCR and long template PCR were used to detect deletions and to define the size of deletions. Evaluation of fundus changes and ERG, lod score estimation, and identification of a mutation in the GRK1 gene were carried out., Results: All patients had night blindness since early childhood. Irregular coarse pigmentation was observed in the peripheral retina of each patient. The fundus appearance before and after 4 h of dark adaptation was similar except that the peripheral retinal pigmentary changes were slightly less evident after extended dark adaptation. Minimal or no rod function with normal cone function on ERG recordings were detected in all three affected members. The rod showed slow recovery to nearly normal amplitude after 4 h in the dark ERG in one individual but not in two other patients. A genome-wide scan showed linkage only to D13S285. Fine mapping defined a region from D13S1315 to 13qter, with a lod score of 2.89 at theta=0 shown by D13S285 and 2.90 at theta=0 by the D13S261-D13S285-D13S1295-D13S293 haplotype. Analysis of the GRK1 gene, which is included in this interval, identified a c.827+623_883del mutation. This intragenic deletion cosegregates with the disease in the family and is only homozygous in affected individuals. This mutation was not detected in 96 controls., Conclusions: The retinal disease in the family reported here has several features differing from typical Oguchi disease, including an atypical Mizuo-Nakamura phenomenon and a non-recordable rod ERG even after 4 h of dark adaptation. Normal visual acuity, normal caliber of retinal blood vessels, and normal cone response on ERG recording suggest retinal dysfunction rather than degeneration (i.e., a variant form of Oguchi disease but unlikely to be retinitis pigmentosa). The disease in the Pakistani family localizes to 13q34 and is caused by a novel deletion including Exon 3 of the GRK1 gene.

Purpose: To identify the disease locus for autosomal recessive congenital cataracts in consanguineous Pakistani families., Methods: Two Pakistani families were ascertained, patients were examined, blood samples were collected, and DNA was isolated. A genome-wide scan was performed using >382 polymorphic microsatellite markers on genomic DNA from affected and unaffected family members. Two-point lod scores were calculated, haplotypes were formed by inspection, and candidate genes were sequenced. Real-time quantitative PCR techniques were used to determine the mRNA levels, and molecular modeling was performed to gain a better understanding of the significance of the disease-causing mutation., Results: In the genome-wide scan, maximum lod scores of 2.67 and 2.77 for family 60004 and 2.02 and 2.04 for family 60006 were obtained for markers D22S539 and D22S315, respectively. The linked region, 22.7 cM (10 Mb) flanked by markers D22S420 and D22S1163, contains the beta-crystallin gene cluster including the genes CRYBA4, CRYBB1, CRYBB2, and CRYBB3. Sequencing of these genes showed a G-->C transition in exon 6 of CRYBB3 resulting in a p.G165R change in the betaB3-crystallin protein that cosegregates with the disease in both families. Real-time PCR analysis suggested that betaB3-crystallin mRNA levels approximate those of other betagamma-crystallins. Molecular modeling predicted changes in electrostatic potential that would be expected to reduce the stability of the fourth Greek-key motif, and hence the entire protein, dramatically., Conclusions: For the first time, a mutation in CRYBB3 is reported in two consanguineous Pakistani families with autosomal recessive congenital cataracts.