Mapping of genes involved in rheumatoid arthritis in Pakistani patients

Abstracts / Current Opinion in Biotechnology 22S (2011) S15–S152

ples in a total of 200 animals and 200 animal product samples. Meat (n = 40) and milk mixtures (n = 40) from identified animals were also targeted by DNA analyses to discriminate the individuals in these mixtures. Individuals were successfully (100%) detected and traced-back from individual animal products. It has been concluded that established EID/DNA integrated system could provide reliable information, and further studies should conducted in order to expand future approaches, for concurrent traceability of animal products. doi:10.1016/j.copbio.2011.05.098

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duction capacity in addition to showing antimicrobial activity. We made an extensive screen of biosurfactant producing bacteria from different environments. Drop collapse oil test and atomized oil assay were utilized to identify strains showing increased biosurfactant production. Analysis of hemolytic and antimicrobial activity of these strains enabled selection of a strain that produces large amounts of a biosurfactant that has antimicrobial activity against four different pathogen microorganisms.The bacterial strain in our study belongs to a species that is not known to produce biosurfactants, implying that biosurfactant from that strain could be novel. Our preliminary LC–MS studies reinforce this idea. We are currently trying to identify the aforementioned biosurfactant and performing experiments to scale-up the production of this biosurfactant.

Kinetic study of the lactic acid fermentation of cabbage juice with Bifidobacterium sp

doi:10.1016/j.copbio.2011.05.100

Buruleanu Claudia Lavinia, Nicolescu Carmen Leane, Manea Iuliana, Bratu Magda Gabriela, Avram Daniela

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Valahia University of Targoviste, Faculty of Environmental Engineering and Biotechnology, Targoviste, Romania E-mail address: [email protected] (B.C. Lavinia) The study was focused on the suitability of the cabbage juice for Bifidobacterium sp. growth in terms of obtaining functional foods and also of control the factors involved in the total quality of the final products. In the same conditions were subjected to fermentation: cabbage juice, cabbage juice heat-treated, respectively, cabbage juice filtered and heat-treated. Some chemical and microbiological parameters were determined in dynamics: pH, lactic acid, acetic acid, reducing sugars and viable cells count through methods standardized or quoted in literature. The experimental data were analyzed using SPSS Statistics 19. The maximum rate of acidification Vmax ranged between 2.66 and 5.58, while the time to reach pH 5.0 (hours) and the time to complete the fermentation (tpH 4, 2 , hours) were 8 and 23, respectively, in the case of the heat-treated juice. No significant differences concerning the cell viability after 24 hours were registered, this one varying between 9.08 and 9.5 log CFU/ml, while the maximum volumetric productivity was 13.1 × 1010 CFU/l hour for the heat-treated juice. Although the kinetic parameters were average, the cabbage juice untreated by heating, without nutrients addition, can be a favourable environment for obtaining probiotic products, with a balanced sensorial and chemical composition.

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Structure based discovery of small molecules to regulate the activity of insulin degrading enzyme Metin Turkay, Bilal Cakir, Onur Dagliyan, I Halil Kavakli, Seda Kizilel Koc University, Istanbul, Turkey E-mail address: [email protected] (M. Turkay) Insulin-degrading enzyme (IDE) is an allosteric Zn +2 metalloprotease involved in the degradation of many peptides including amyloid beta (A␤), and insulin that play key roles in Alzheimer’s disease (AD) and type 2 diabetes mellitus (T2DM), respectively. Crystal structure of IDE revealed that N-terminal of IDE has an exosite which is ∼30 A˚ away from the catalytic region and serves as a regulation site by orientation of the substrates of IDE to the catalytic site. In this study, we applied structure based drug design methodology to discover novel small molecule organic compounds that enhance the activity of human IDE. The activity of novel compounds were tested using in vitro essays by enhanced IDE mediated proteolysis of substrate V, insulin and FA␤B degradation, respectively. These compounds demonstrated submicromolar activation. In addition, amino acid mutations at the exosite of IDE verifies that the designed molecules bind to the targeted area. This study describes the first examples of a computer-aided discovery of IDE regulators, showing that in vitro activation of this important enzyme with small molecules is possible.

doi:10.1016/j.copbio.2011.05.099

doi:10.1016/j.copbio.2011.05.101

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Identification and characterization of a biosurfactant from a novel bacterial strain

Mapping of genes involved in rheumatoid arthritis in Pakistani patients

Turgay Tekinay, Ozgun Candan Onarman, Diren Han

Attya Bhatti 1 , Peter John 1 , Hajra Sadia 1 , Iltaf Ahmed 1 , Ishtiaq Qadri 1 , Fazal Jalil 1 , Zeeshan Gauhar 2 , Moghees Ikram 2 , Javaid Malik 3 , Mushtaq Ahmed 4

Bilkent University, UNAM Institute of Materials Science Nanotechnology, Ankara, Turkey

1

E-mail address: [email protected] (T. Tekinay) Biosurfactants are surface active compounds produced by microorganisms. Their advantages such as biodegradability and lower toxicity make them superior to their chemical counterparts. Biosurfactants could be used in different industries including biomedicine, bioremediation, food, pharmaceutical and cosmetics applications. However, large scale production of many type of biosurfactants is limited due to their high production cost. We aimed to isolate a new bacterial strain that has a high biosurfactant pro-

National University of Sciences and Technology (NUST), Islamabad, Pakistan 2 KRL Hospital, Islamabad, Pakistan 3 Fauji Foundation Hospital, Rawalpindi, Pakistan 4 MH Hospital, Rawalpindi, Pakistan E-mail address: [email protected] (A. Bhatti) Rheumatoid arthritis (RA) is a chronic auto-immune disease with extensive articular cartilage destruction. RA results from interactions between genetic and environmental factors. The study

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Abstracts / Current Opinion in Biotechnology 22S (2011) S15–S152

involved clinical and genetic evaluation of 350 sporadic RA patients from rheumatology units of Rawalpindi and Islamabad hospitals and a linkage analysis of two Pakistani families (A and B) with autosomal recessive RA from different regions of Pakistan. Descriptive Statistical analysis showed that RA preferentially affects women with female to male ratio of 2.8:1 but equal rates are seen in females and males at 60–75 age groups. Similarly, RA is more prevalent between 45 and 59 years age group, in female 40 yrs and in male 50 yrs is the peak age (p = 0.004). The linkage analysis of two families A and B was performed by genotyping with highly polymorphic microsatellite markers corresponding to known loci. Genetic linkage revealed that affected individuals were heterozygous for different combinations of parental alleles, thus indicating exclusion of families to known loci of RA suggesting involvement of a novel locus in the onset of disease and genome wide search will be carried out. Identification of RA genes will provide new insights into disease pathogenesis and finally will generate potential targets for therapy. doi:10.1016/j.copbio.2011.05.102

OP32 The incidence of Y-chromosome microdeletions in Pakistani infertile men Nosheen Mujtaba 1 , Mamoona Naz 2 , Saima Perveen 1 , Mamoona Yasmin 1 , Fida Haider 1 , Rubina T Siddiqui 2 1

National Institute for Biotechnology and Genetic Engineering, Faisalabad, Pakistan 2 Institute of Biochemistry and Biotechnology, University of the Punjab, Lahore, Pakistan E-mail address: [email protected] (N. Mujtaba)

The aim of this study is to investigate the contribution of microdeletions in Y-chromosome in azoospermic and oligozoospermic infertile Pakistani men. A total of 163 males (113 infertile 50 fertile) were included in the study. Five multiplex PCRs targeting AZFa, AZFb, and AZFc regions on Y-chromosome were performed detecting 26 sequence tagged sites (STS) including sY81, sY130, sY141, sY142, sY4, sY155, sY157, sY160, sY182, sY231, sY202, sYPR3, sY126, sY133, sY132, sY153, sY152, sY124, sY86, sY127, sY134, sY254, sY84, sY255, sY72 and CDY2. Individuals with severe oligozoospermia showed a 2.86% deletion frequency in AZFc region and 20% in the AZFb region, as compared to azoospermic males (5.5% in AZFc and 9.25% in AZFb region), whereas moderate oligospermic individuals showed AZFb microdeletion in 12.5% men. None of the individuals from control group showed any microdeletion in Y-chromosome. Though microdeletions in Y chromosome do contribute towards male infertility, other factors such as infections might be playing an important part. Because men with severe infertility suffer a high risk of Y chromosome deletion, screening for these men is recommended prior to treatment with assisted reproduction. doi:10.1016/j.copbio.2011.05.103

OP33 Polymorphism in MECP2 and PAK3 genes in intellectually disabled children Fida Haider 1 , Rubina Tabassum Siddiqui 1 , Asma Arshad Bajwa 2 , Samreen Afzal 2 , Nosheen Mujtaba 1 , Mamoona Yasmin 1 , Imtiaz Ahmad Dogar 2 1 National Institute for Biotechnology and Genetic Engineering, P.O. Box#577, Jhang Road, Faisalabad, Pakistan 2 Punjab Medical College, Faisalabad, Pakistan

E-mail address: [email protected] (F. Haider) To provide insight into the distribution and frequency of mutations in MECP2 and PAK3 genes involved in neuronal development and synaptic plasticity. Exons of MECP2 and PAK3 genes were PCR amplified and mutations were detected by PCR-RFLP, SSCP and DNA sequencing in 100 intellectually disabled children. Cytogenetic analysis was carried out to exclude mental retardation cases associated with chromosomal disorders. Polymorphism in three exons of MECP2 gene was found to be 10%. The A140V mutation commonly associated with Rett syndrome and X-linked mental retardation was absent in the group under investigation. Mutations in exon1 were missense and silent mutations, while exon3E had only missense mutations. Only one nucleotide (T) insertion was found in Exon3C, which resulted in expression of truncated MECP2 protein. The PAK3 exon10 did not show any polymorphism. Intellectual disability is the most prevalent birth-related problem in Pakistan. The genetic basis of intellectual disability is quite complex and needs to be explored in different populations. Our data confirms the involvement of MECP2 gene in brain function but still leaves a room for further research to know the cause of mental retardation in 90% of the cases under study. doi:10.1016/j.copbio.2011.05.104

OP34 The increasing importance of Medical Genetics in Turkey Seher Polat, Sevda Yesim Karabulut, Oguzhan Bahadir, Munis Dundar Medical Genetics Department, Erciyes University, Kayseri, Turkey E-mail address: sp [email protected] (S. Polat) Turkey located at the crossroads between Europe and Asia with a population of about 74 million. It possessed great ethnical diversity. The frequency of consanguineous marriages ranged from 11,5 % to 46% overall in the eastern part and the ratio decreased to 11.7% in the western parts of the country. To draw the attention of the people on increasing importance of genetic diseases, we collected patients diagnosed with mental retardation, growthdevelopmental delay, syndromic, epilepsy in pediatric clinics, infertile woman, habitual aborts, thrombophilia in gynecology clinics, and infertile man in urology clinics between the years of 2005–2010 at Erciyes University Medical Faculty Hospital. 13, 2 % (15673), 1 %, 1 % (1337), 0, 01% (15) and 0,005% (6) diagnosed with growth-developmental delay, epilepsy, syndromic, and mental retardation out of 118, 349 patients, 7, 7% (8831) diagnosed with infertile man out of 115, 028 patients, 7, 5% (13071), 1, 4% (2451) and 0, 3% (557) infertile woman, habitual aborts and thrombophilia out of 173.287 patients, respectively. With recognizing increasing importance of congenital and genetically determined disorders in the world, taking steps to establishing programme and implementing activities to prevent and control these genetic diseases and to