Spondylocarpotarsal synostosis syndrome due to a novel loss of function FLNB variant : a case report

BackgroundLoss of function or gain of function variants of Filamin B (FLNB) cause recessive or dominant skeletal disorders respectively. Spondylocarpotarsal synostosis syndrome (SCT) is a rare autosomal recessive disorder characterized by short stature, fused vertebrae and fusion of carpal and tarsal bones. We present a novel FLNB homozygous pathogenic variant and present a carrier of the variant with short height.Case presentationWe describe a family with five patients affected with skeletal malformations, short stature and vertebral deformities. Exome sequencing revealed a novel homozygous frameshift variant c.2911dupG p.(Ala971GlyfsTer122) in FLNB, segregating with the phenotype in the family. The variant was absent in public databases and 100 ethnically matched control chromosomes. One of the heterozygous carriers of the variant had short stature.ConclusionOur report expands the genetic spectrum of FLNB pathogenic variants. It also indicates a need to assess the heights of other carriers of FLNB recessive variants to explore a possible role in idiopathic short stature.Peer reviewe

A Frameshift Mutation in GRXCR 2 Causes Recessively Inherited Hearing Loss

More than 360 million humans are affected with some degree of hearing loss, either early or later in life. A genetic cause for the disorder is present in a majority of the cases. We mapped a locus ( DFNB 101) for hearing loss in humans to chromosome 5q in a consanguineous P akistani family. Exome sequencing revealed an insertion mutation in GRXCR 2 as the cause of moderate‐to‐severe and likely progressive hearing loss in the affected individuals of the family. The frameshift mutation is predicted to affect a conserved, cysteine‐rich region of GRXCR 2, and to result in an abnormal extension of the C ‐terminus. Functional studies by cell transfections demonstrated that the mutant protein is unstable and mislocalized relative to wild‐type GRXCR 2, consistent with a loss‐of‐function mutation. Targeted disruption of G rxcr2 is concurrently reported to cause hearing loss in mice. The structural abnormalities in this animal model suggest a role for GRXCR 2 in the development of stereocilia bundles, specialized structures on the apical surface of sensory cells in the cochlea that are critical for sound detection. Our results indicate that GRXCR 2 should be considered in differential genetic diagnosis for individuals with early onset, moderate‐to‐severe and progressive hearing loss. We mapped a new deafness locus DFNB101 and discovered an insertion mutation in GRXCR2 as the cause of moderate‐to‐severe hearing loss in humans. The frameshift mutation was predicted to affect the conserved, cysteine‐rich domain of GRXCR2, and to result in an abnormal extension of the C‐terminus. Cell transfection experiments demonstrated that the mutant protein is unstable and mislocalized relative to the wild type GRXCR2, consistent with a loss of function mutation.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/106992/1/humu22545.pd

Autosomal recessive chondrodysplasia with severe short stature caused by a biallelic COL10A1 variant

Background Heterozygous mutations in COL10A1 underlie metaphyseal chondrodysplasia, Schmid type (MCDS), an autosomal dominant skeletal dysplasia. Objective To identify the causative variant in a large consanguineous Pakistani family with severe skeletal dysplasia and marked lower limb deformity. Methods Whole exome sequencing was completed followed by Sanger sequencing to verify segregation of the identified variants. In silico variant pathogenicity predictions and amino acid conservation analyses were performed. Results A homozygous c. 133 C>T (p.Pro45Ser) variant was identified in COL10A1 in all six severely affected individuals (adult heights 119-130 cm, mean similar to -6.33 SD). The individuals heterozygous for the variant had mild phenotype of short stature (adult heights 140-162 cm, mean similar to -2.15 SD) but no apparent skeletal deformities. The variant was predicted to be pathogenic by in silico prediction tools and was absent from public databases and hundred control chromosomes. Pro45 is conserved in orthologues and is located in the non-collagenous 2 domain of COL10A1, variants of which have never been associated with skeletal dysplasia. Conclusions This first report of individuals with a homozygous variant in COL10A1 defines a new type of autosomal recessive skeletal dysplasia. The observations in COL10A1 variant carriers suggest a phenotypic overlap between the mildest forms of MCDS and idiopathic short stature.Peer reviewe

PHYTOCHEMICAL ANALYSIS AND IN VITRO BIOLOGICAL CHARACTERIZATION OF AQUEOUS AND METHANOLIC EXTRACT OF BACOPA MONNIERI

Objective: The present study was designed to identify the phytocompounds, to compare the antibacterial, antioxidant, and anti-inflammatory effects of aqueous and methanolic extract of Bacopa monnieri.Methods: Antioxidant activity was determined by 1, 1-diphenyl-2-picrylhydrazyl (DPPH), Ferric Reducing Antioxidant Power (FRAP), Super oxide dismutase (SOD), Reduced glutathione (GSH), Catalase assays. Anti-inflammatory activity was measured with inhibition of albumin denaturation and trypsin inhibitory assay. Finally, extracts were tested against various pathogenic bacterial and fungal strains by broth dilution assay and disc diffusion assay respectively.Results: Results showed the presence of alkaloids, flavonoids, phenols, quinines and glycosides etc while steroids and carboxylic acid were absent. The extracts demonstrated free radical-scavenging activity quite comparable with standard ascorbic acid. Methanolic extract exerted comparative higher antioxidant and anti-inflammatory activity than aqueous extract. Both extracts were most effective against Bacillus subtilis and lowest inhibition against Staphylococcus aureus.Conclusion: The results obtained clearly indicated a promising potential of B. monnieri for serving as a strong ROS scavenger, might be used as anti-arthritic and strong natural antibiotic agent for effective treatment of various oxidative stressed disorders (cancer, cardiovascular diseases), inflammatory disorders (rheumatoid arthritis) and various bacterial infections

Van Der Knaap Disease in a 3-year-old Male Child: A Case Report

Van der Knaap disease or megalencephalic leukoencephalopathy with subcortical cysts is a leukodystrophy with autosomal-recessive inheritance caused by mutation in the gene MLC1 which is localized on chromosome 22q. It is characterized by macrocephaly, motor developmental delay, seizures, spasticity, ataxia, and mild mental deterioration. On neuroimaging, involvement of cerebral white matter along with subcortical cysts in frontal and temporal lobes are hallmarks of the disease. There is no definite treatment of this disease. We report a case of Van Der Knaap disease in a 3-year-old male child who presented with seizures and delayed developmental milestones

Enhanced production of subtilisin of Pyrococcus furiosus expressed in Escherichia coli using autoinducing medium

A subtilisin gene identified in the reported genome sequence of Pyrococcus furiosus was amplified and inserted in pET-22b(+) vector to produce the recombinant plasmid pET-SB. Escherichia coli BL-21 (DE3)CodonPlus was transformed with this plasmid and the enzyme was expressed up to 30% of the total cell protein on induction with IPTG. The expressed protein appeared at a position corresponding to ~20 kDaon SDS-PAGE as compared to theoretical molecular mass of 17.6 kDa. This aberrant electrophoresis mobility could be due to specific amino acid composition of the protein. Auto-induction with lactose also produced a similar level of expression but the total amount of the enzyme produced was 2.4 foldgreater than that when produced with IPTG induction. This was due to a higher cell density obtainable in the auto-inducing medium. The enzyme expressed in the insoluble state could be partially refolded after denaturation with urea at high pH. This study reports for the first time high-level expression ofsubtilisin of P. furiosus in E. coli using an auto-inducing medium

Natural and Anthropogenic Sources of Groundwater Salinization in Parts of Karachi, Pakistan

This study is aimed to evaluate the groundwater quality of Gulshan-e-Iqbal and Liaquatabad towns inKarachi. Thirty (n=30) groundwater samples were randomly collected from different locations by electrically pumpedwells at various depths (14-91m). All the water samples were analyzed to determine their suitability for drinkingpurpose based on various physicochemical parameters. Data reveal that high concentration of TDS and hardness havedeteriorated the groundwater quality of study area. The main phenomenon responsible for groundwater pollution is theseawater intrusion due to the proximity of study area to the Arabian sea. Large scale unplanned urbanization, poorwaste management and other anthropogenic activities have also triggered the deterioration of groundwater quality.Study showed that local geology plays vital role in the distribution of major cations and anions. Data suggested thatground water of this study area is highly contaminated by seawater intrusion and considered not fit for drinkingpurpose

ASSESSING THE THERAPEUTIC ROLE OF JOSHANDA: PHYTOCHEMICAL, ANTIOXIDANT, ANTI-INFLAMMATORY AND ANTIMICROBIAL ACTIVITIES

Objective: Joshanda, a polyherbal Unani formulation is extensively used as a common home remedy for the treatment of a cough and cold accompanied by pharyngeal inflammation and fever. This study aimed to analyze phytochemicals, antioxidant, anti-inflammatory, and antibacterial activity.Methods: The study investigated the presence of phyto-compounds in joshanda and antioxidant, antibacterial, anti-fungal, and anti-inflammatory activities by various in vitro standard methods using ascorbic acid, ampicillin, and aspirin respectively as standard drugs.Results: Joshanda aqueous extract revealed the presence of tannins, phenols, flavonoids glycosides, terpenoids, and alkaloids and absence of sterols, saponins, xanthoprotiens, and carboxylic acid. Joshanda showed the highest inhibition against B. subtilis (% MGI 99.000±0.577) and least inhibition against P. aeruginosa (%MGI of 84.102±0.491). Joshanda extract, ascorbic acid demonstrated highest % DPPH radical scavenging of 98.379±0.313%, 98.843±0.443% and a minimum of 36.210±1.174%, 83.192±0.422%. Results showed H2O2 scavenging activity of 0.047±0.001 μg/ml per minute degradation of H2O2. FRAP value was observed in joshanda and ascorbic acid with a maximum of 0.945±0.024, 0.687±0.047 mmol and minimum of 0.171±0.036, 0.059±0.005 mmol respectively. Joshanda extract showed the highest albumin denaturation inhibition of 14.069±0.350% and the lowest of 1.880±0.194% at extract volume of 1000 µl and 100 µl respectively. The extract demonstrated the highest proteinase inhibition of 24.003±0.291 % and the lowest of 4.959±0.254% comparable to aspirin. Joshanda had no potent anticandidal activity up to 1 mg/ml.Conclusion: Results clearly suggested that joshanda is a potent phytodrug and can also be used as a strong reactive oxygen species scavenger, might be used as anti-arthritic and strong natural antibiotic agent for effective treatment of various oxidative stressed disorders

Myeloperoxidase and elastase are only expressed by neutrophils in normal and in inflammed liver

Myeloperoxidase (MPO) is involved in acute and chronic inflammatory diseases. The source of MPO in acute liver diseases is still a matter of debate. Therefore, we analysed MPO-gene expression on sections from normal and acutely damaged [carbon tetrachloride-(CCl4) or whole liver γ-Irradiation] rat liver by immunohistochemistry, real time PCR and Western blot analysis of total RNA and protein. Also total RNA and protein from isolated Kupffer cells, hepatic stellate cells, Hepatocytes, endothelial cells and neutrophil granulocytes (NG) was analysed by real time PCR and Western blot, respectively. Sections of acutely injured human liver were prepared for MPO and CD68 immunofluorescence double staining. In normal rat liver MPO was detected immunohistochemically and by immunofluorescence double staining only in single NG. No MPO was detected in isolated parenchymal and non-parenchymal cell populations of the normal rat liver. In acutely damaged rat liver mRNA of MPO increased 2.8-fold at 24 h after administration of CCl4 and 3.3-fold at 3 h after γ-Irradiation and MPO was detected by immunofluorescence double staining only in elastase (NE) positive NGs but not in macrophages (ED1 or CD68 positive cells). Our results demonstrate that, increased expression of MPO in damaged rat and human liver is due to recruited elastase positive NGs