Apparent Missense Variant in COL7A1 Causes a Severe Form of Recessive Dystrophic Epidermolysis Bullosa via Effects on Splicing.

Dystrophic epidermolysis bullosa is an inherited skin disorder characterized by fragile skin that is prone to blistering. We report here a consanguineous Pakistani family with two siblings, in whom a severe recessive dystrophic epidermolysis bullosa was suspected. Using whole-exome sequencing for one sibling, the homozygous base substitution c.7249C>G in COL7A1 was identified, and could be confirmed in the other sibling by Sanger sequencing. In our exome data, this mutation was annotated as a missense substitution (p.Gln2417Glu), but in silico tools indicated a possible effect on splicing. Using the ExonTrap vector it was verified that the mutation leads to activation of a cryptic donor splice site, which leads to loss of 26 nucleotides, and a frame-shift event predicted to result in a truncated protein (p.Q2417Sfs*57). The present report de-scribes an apparent COL7A1 missense substitution with an unexpected consequence on splicing that leads to a severe recessive dystrophic epidermolysis bullosa phenotype.

In Vitro Anti-Inflammatory, Anticancer (MCF-7, 3T3, and HeLa Cell Lines), and Brine Shrimp Lethality Assay and FTIR Analysis of the Extract and Fractions of the Whole Plant of Heliotropium europaeum.

In this study, anti-inflammatory, anticancer, brine shrimp lethality, and FTIR studies were evaluated. The oxidative burst assay using the chemiluminescence technique, MTT assay, brine shrimp lethality assay, and FTIR analysis were the methods used for the evaluation of anti-inflammatory, anticancer, brine shrimp lethality, and FTIR studies, respectively. The whole-plant butanol fraction of Heliotropium europaeum (WBFHE) showed anti-inflammatory activity on ROS having IC5014.7 ± 2.5 while the extract and other fractions of the whole plant of Heliotropium europaeum exhibited no anti-inflammatory activity. None of the extract and fractions of the whole plant of Heliotropium europaeum exhibited anticancer (MCF-7, 3T3, and HeLa cell lines) activities. The whole-plant aqueous fraction of Heliotropium europaeum (WAFHE) and whole-plant butanol fraction of Heliotropium europaeum (WBFHE) showed lethality at high concentration while at low concentration, no toxicity was shown. The whole-plant methanolic extract of Heliotropium europaeum (WMEHE) and whole-plant n-hexane fraction of Heliotropium europaeum (WHFHE) exhibited no toxicity. FTIR interpretation showed the functional groups for the aromatic compounds, phenols, carboxylic acids, esters, alkanes, alkenes, alcohols, alkyl halides, sulfate esters, phosphines, silanes, nitriles, thiols, amines, phosphoric acids, and nitro compounds.

Comparative analysis of antioxidant activity, toxicity, and mineral composition of kernel and pomace of apricot (Prunus armeniaca L.) grown in Balochistan, Pakistan.

The present study aims to investigate some physical attributes, total phenolics content, total flavonoids content, mineral composition, bioluminescence toxicity assay and antioxidant activity in terms of DPPH, HPS, TAC and FRAP assays in the kernel and pomace samples of six apricot cultivars grown in Balochistan, Pakistan. TFC and TPC determined by the AlCl3 and Folin-Ciocalteu assays in apricot kernel extracts of six cultivars varied from 1797.5 (Chagali) to 4778.9 (Badoghur) mg QUE/100 g DW and from 1750.0 (Chagali) to 5005.8 (Badoghur) mg GAE/100 g DW. Apricot kernels exhibited higher antioxidant activity than pomace; antioxidant activity in terms of IC50 in kernels ranged from 24.88 to 98.61 µg/ml for DPPH, 334.84 to 516.63 µg/ml for HPS, from 22.02 to 110.80 µg/ml for TAC and from 96.27 to 163.35 µg/ml for FRAP. The apricot kernels showed higher TPC, TFC, bioluminescence toxicity to V. logei and antioxidant activity than the pomace. The correlation analysis demonstrated substantial contributions of polyphenols and flavonoids to antioxidant assays. The sample type was the leading factor affecting the amounts of K, Na, Ca, Fe, and Mn in the tested samples; mineral contents were higher in pomace than kernels. The highest inhibition to V. logei was found in the kernels of Badoghur (IC50 = 1.61 mg/ml). The PCA analysis showed significant contributions of phenolic and flavonoid contents towards antioxidant bioluminescence toxicity assays. Our results suggest Badoghur, Shakarpara and Sardai kernels are rich sources of secondary metabolites and possess remarkable antioxidant and antiluminescence activity and can make a significant contribution to the treatment and prevention of chronic health problems.