Shagufta Naz

In vitro conservation and production of vigorous and desiccate tolerant synthetic seeds in Stevia rebaudiana

cultures for encapsulation. The explants were taken from green houses and tissue culture laboratory of AgriBiotech Research Farms, Lahore, Pakistan. Different concentrations of sodium alginate and calcium chloride affect the shape and germination of artificial seeds. A gelling matrix of 3% sodium alginate and 100 mM calcium chloride was found most suitable for the formation of firm, clear and isodiametric ideal beads. No effect of the duration of sodium alginate treatment on bead formation was observed. However, timing of CaCl

Initiation, proliferation and development of micro-propagation system for mass scale production of banana through meristem culture

Explants were taken from field grown plants in 2010. The shoot apical meristem of different sizes was cultured on Murashige and skoog’s (MS) medium supplemented with different concentrations and combinations of 6-benzylamino-purine (BAP), kinetin (Kin) and α- naphthaleneacetic acid (NAA) either alone or in combination with each other under different temperature conditions ranging from 23 to 27°C. Shoot formation response from shoot apical meristem showed that MS medium containing 1.0 mg/l BAP showed best response for shoot formation. For shoot multiplication, MS medium containing 1.0 mg/l BAP + 0.25 mg/l kin provided the best multiplication response which was 8 shoot per culture vial within 21.6 days after inoculation into shoot multiplication medium. Shoot formation and multiplication response was also affected by temperature variations. The best results were obtained at 27°C ± 1°C. By increase or decrease in temperature, the rate of in vitro response was also decreased. For rooting of well developed in vitro shoots MS medium supplemented with 1.0 mg/l Indole-3- butyric acid (IBA)+ 0.5 mg/l NAA showed 3.6 roots per plant after 6.8 days of inoculation into rooting medium with an average root length of 2.4 cm. 100% hardening response was obtained in Peat moss after 21 days of transplantation in glass house. The experiments were designed in completely randomized pattern. Key words: Murashige and skoog’s medium, proliferation, banana, micro propagation system.

Enhanced production of xylanase from locally isolated fungal strain using agro-industrial residues under solid-state fermentation

This study is related to the isolation of fungal strain for xylanase production using agro-industrial residues. Forty fungal strains with xylanolytic potential were isolated by using xylan agar plates and quantitatively screened in solid-state fermentation. Of all the tested isolates, the strain showing highest ability to produce xylanase was assigned the code Aspergillus niger LCBT-14. For the enhanced production of the enzyme, five different fermentation media were evaluated. Out of all media, M4 containing wheat bran gave maximum enzyme production. Effect of different variables including incubation time, temperature, pH, carbon and nitrogen sources has been investigated. The optimum enzyme production was obtained after 72 h at 30°C and pH 4. Glucose as a carbon source while ammonium sulphate and yeast extract as nitrogen sources gave maximum xylanase production (946 U/mL/min). This study was successful in producing xylanase by A. niger LCBT-14 economically by utilising cheap indigenous substrate.

ABO blood group frequency in Ischemic heart disease patients in Pakistani population.

Objectives: To determine if there is any significant association between ABO blood groups and ischemic heart disease (IHD). Methods: The study was performed at Punjab Institute of Cardiology (PIC), Lahore. Study duration was from January 2012 to September 2012. This study included 200 IHD patients and 230 control individuals. Self design questionnaire was used to collect information regarding risk factors. Standard agglutination test was performed to determine the blood groups. Data was analyzed on SPSS 16. Results: The prevalence of blood groups in IHD group was 34% in blood group A, 29% in blood group B, 14% in blood group AB and 23% in blood group O. In control group the distribution of B, A, AB and O blood groups were 34.4%, 20.9%, 12.6%, 32.2% respectively. Rh+ve factor was prevalent in 90.5% among IHD group and 92.6% in control subjects. The prevalence of IHD was more in males (63.5%) as compared to females (36.5%). Mean age was 56.4±0.86 (yrs) and BMI was 26.4±0.33 (kg/m2). The prevalence of hypertension was 58.5%, diabetes was 53%, family history of cardiac disease was 45%, 35.5% of patients were doing exercise regularly, 58.5% used ghee, and 58% were smokers. Conclusion: Subjects with blood group A had significantly (p< 0.05) higher risk of developing IHD as compare to other blood groups.

Bioconversion potential of Trichoderma viride HN1 cellulase for a lignocellulosic biomass Saccharum spontaneum

The industrialisation of lignocellulose conversion is impeded by expensive cellulase enzymes required for saccharification in bioethanol production. Current research undertakes cellulase production from pretreated Saccharum spontaneum through Trichoderma viride HN1 under submerged fermentation conditions. Pretreatment of substrate with 2% NaOH resulted in 88% delignification. Maximum cellulase production (2603 ± 16.39 U/mL/min carboxymethyl cellulase and 1393 ± 25.55 U/mL/min FPase) was achieved at 6% substrate at pH 5.0, with 5% inoculum, incubated at 35°C for 120 h of fermentation period. Addition of surfactant, Tween 80 and metal ion Mn+2, significantly enhanced cellulase yield. This study accounts proficient cellulase yield through process optimisation by exploiting cheaper substrate to escalate their commercial endeavour.

Homozygous p.G61E mutation in a consanguineous Pakistani family with co-existence of juvenile-onset open angle glaucoma and primary congenital glaucoma.

Glaucoma is one of the primary causes of visual impairment and blindness in the world. It is characterized by the damage to the optic nerve head and visual field loss. Variants in CYP1B1 are the most common cause of glaucoma in different world populations. We studied a consanguineous Pakistani family in which three affected individuals had a severe form of glaucoma with members in one generation diagnosed with juvenile-onset open angle glaucoma at 27 years of age, while the members of the next generation were affected with primary congenital glaucoma with onset at birth. Sequencing of CYP1B1 revealed a homozygous transition variant, c.182G>A, p.G61E which co-segregated with the disease phenotype. This variant has been previously reported to cause both recessively and dominantly inherited PCG and JOAG in different populations. However, this reported for the first time in Pakistani PCG and JOAG patients in a homozygous state. This is also the first ever report of a CYP1B1 variant segregating in a consanguineous family with co-existence of JOAG and PCG in two subsequent generations. This observation of different phenotypes due to an identical mutation suggests that primary congenital glaucoma and juvenile-onset open angle glaucoma can both be caused by homozygosity for the same mutation. It also indicates the reduced penetrance of the variant in those affected due to p.G61E mutation and further implies that modifiers have a role in controlling the time of onset of the disorder.

Evaluation of antifungal and antioxidant potential of two medicinal plants: Aconitum heterophyllum and Polygonum bistorta

ABSTRACT Objective To focus on the evaluation of antimicrobial and antioxidant activity of two endangered medicinal plants Aconitum heterophyllum (A. heterophyllum) and Polygonum bistorta (P. bistorta). Materials Plant extracts were obtained by using microwave assisted extraction method. The in vitro antifungal activity of A. heterophyllum and P. bistorta extracts were determined by measuring diameters of inhibitory zones of these extracts against Aspergillus niger and Alternalia solani. Results Methanolic extract of A. heterophyllum showed significant (P≤0.05) antifungal activity against both the tested organisms. It was also observed that ethanolic extracts of P. bistorta also had good antifungal activity against the tested fungal strains as compared to the methanolic extracts. It showed significant antifungal activity (P≤0.05) against both the tested strains. Antioxidant activity of methanolic and ethanolic extracts of A. heterophyllum and P. bistorta were also measured using a radical scavenging method. Ascorbic acid was used as a standard. Conclusions It was observed that A. heterophyllum and P. bistorta have significant antioxidant activity. Higher antioxidant activity was recorded in methanolic extract of A. heterophyllum as compared to its ethanolic extract. However, in case of P. bistorta ethanolic extract of the plant exhibited higher antioxidant potential than methanolic extracts. Hence both of these plants have significant antimicrobial as well as antioxidant potential.

In Vivo Crystallization of Three-Domain Cry Toxins

Bacillus thuringiensis (Bt) is the most successful, environmentally-friendly, and intensively studied microbial insecticide. The major characteristic of Bt is the production of proteinaceous crystals containing toxins with specific activity against many pests including dipteran, lepidopteran, and coleopteran insects, as well as nematodes, protozoa, flukes, and mites. These crystals allow large quantities of the protein toxins to remain stable in the environment until ingested by a susceptible host. It has been previously established that 135 kDa Cry proteins have a crystallization domain at their C-terminal end. In the absence of this domain, Cry proteins often need helper proteins or other factors for crystallization. In this review, we classify the Cry proteins based on their requirements for crystallization.

Molecular characterisation of Bacillus chitinase for bioconversion of chitin waste

In this work chitin was extracted chemically from shrimp shells. Seventeen Bacillus isolates were screened for chitinolytic activity. The chitinolytic strains of Bt. were screened at different temperatures and pHs for their hydrolytic potentials. By using a pair of specific primers, endochitinase gene was amplified from SBS Bt-5 strain through PCR, and then cloned into pTZ57 TA cloning vector and transferred in Escherichia coli DH5α strain. The sequenced gene (GenBank Accession No: HE995800) consists of 2031 nucleotides capable of encoding 676 residues. The protein consisted of three functional domains with a calculated molecular mass of 74.53 kDa and a pI value of 5.83. The amino acid sequence of chi gene showed 99% similarity to the genes of Bt MR11 endochitinase, Bt serovar kurstaki chitinase (kchi), Bt strain MR21 endochitinase and Bacillus cereus B4264.

Process optimisation for the biosynthesis of cellulase by Bacillus PC-BC6 and its mutant derivative Bacillus N3 using submerged fermentation

This study deals with optimisation of cultural conditions for enhanced production of cellulase by Bacillus PC-BC6 and its mutant derivative Bacillus N3. Influence of different variables including incubation time, temperature, inoculum size, pH, nitrogen sources and metal ions has been studied. The optimum conditions for cellulase production were incubation period of 72 h, inoculum size 4% incubation temperature 37°C, pH 7, 0.25% ammonium sulphate, 0.2% peptone as inorganic and organic nitrogen source in case of Bacillus PC-BC6. In case of mutant Bacillus N3, optimal conditions were incubation period of 48 h, incubation temperature 37°C, inoculum size 3%, pH 7, 0.2% ammonium chloride and 0.15% yeast extract. Presence of MnSO4 and CaCl2 enhances the enzyme production by Bacillus PC-BC6 and mutant Bacillus N3, respectively. This study was innovative and successful in producing cellulase economically by using cheap indigenous substrate Saccharum spontaneum.