The main objectives for this study were to determine the molecular basis of the disease resulting in the particular clinical phenotype of thalassemia intermedia in Pakistan, to identify the factors affecting genotype – phenotype relationship, to determine the possibility for a consistent prediction of phenotype severity from the genotype factors and to asses their relative importance. Thalassemia Intermedia is clinically and genetically heterogeneous and the genotype is retrospective. However, the disease being of milder form is characterized by late commencement of transfusion, lesser degree of anemia and greater survival time. In this study one consistent factor having a fair ground for the classification was the age of the patient at presentation. Age of the patients in this study was between 2.5 – 36 years. These patients in the radiance of presentation were grouped in four categories; Severe, Moderate, Mild and Very Mild depending on the transfusion commencement. Eleven different beta chain mutations were identified, IVSI-5 = 46 %, Fr 8-9 = 11.5 %, Cap+1 = 10%,, Cd30 = 7.0%, IVSI-I 6.5%, HbE = 6%, HbS = 3%, Del 619 = 1.5 %, Cd15 = 1.0%, Fr 41 – 42 = 0.5%, Fr16 = 0.5% and δβ = 5%. However, 1.5 % of the alleles remained unknown. Out of 100 samples tested for Xmn-I polymorphism 79 were found to be positive, 36 % for +/+ genotype and 43% for -/+ genotype and 21% were negative for the genotype. The samples were also tested for δβ mutations and 5 of them were found to be homozygous. Deletions for α- chain were observed in 30% of the samples, all of them had α3.7 deletions out which 8 % had - α/ - α deletions , 21% had -α/αα deletions and 1% had - - / - α deletions (Table 3.46). Frequency of Alpha Thalassemia in different ethnic groups were determined which revealed that alpha thalassemia mutations are more prevalent in Sindhis, Punjabis and Mahajirs. Relating phenotype to genotype is complicated by complex interaction of the environment and other genetic factors such as coinheritance of other hemoglobin mutations. Alpha thalassemia and Xmn-I predominantly contributed the phenotype. Hemoglobinopathies account for only 9% of the patients. Compound heterozygosity was another factor involved particularly with the assistance of Xmn-I polymorphism and coexistence of α- Thalassemia. Xmn-I +/+ and Hemoglobin S mutation accounted for 9% of cases. To establish a comprehensive diagnosis program the problem of detection of an ability to produce fetal hemoglobin, inheritance of β+ Thalassemia genes and inheritance of α Thalassemia and other factors ameliorating the disease should be defined and incorporated. Molecular basis of thalassemia intermedia as defined in this study explains the involvement of different factors that tend to develop the disease phenotype. However, no single factor finds an authority for the discipline of mildness and thus require cooperation of the elements serving in amelioration.
The Immensely Merciful to all, The Infinitely Compassionate to everyone.
35:01 a. The Praise and Gratitude is for Allah – The One and Only God of everyone and everything, - Creator of the celestial realm and the terrestrial world without any precedent, - The Appointer of the angels as message-bearers, with two and three and four pairs of wings. b. HE increases creation as and what HE Wills. c. Indeed, Allah Manifests Sovereignty over all existence.
35:02 a. Whatever mercy and good fortune Allah may open up for a people, no one can or is able to hold it back, and b. whatever of these HE may hold back, there is not one, who can or is able to give it after HIM deciding not to give, c. for HE is The Almighty, The Wise.
35:03 a. O The People of the World! b. Remember Allah’s blessings and favors upon you - c. Is there any creator, other than Allah, who can provide for you sources of sustenance from the sky and the earth? d. No. There can never be one! e. There is no entity of worship –and can never be - apart from HIM. f. How, then, can you be so self-deceiving?
35:04 a. And if they belie and deny you, O The Prophet, know that Messengers before you were also belied and denied. b. And ultimately all matters are to be referred to Allah for resolution.
35:05 a. O The People of the World! b. Indeed, Allah’s Promise about the Hereafter is true, c. therefore, do not be seduced by the worldly life,
Watermelon is gaining importance as a functional food due to its therapeutic effect. The therapeutic effect of watermelon has been reported and has been attributed to antioxidant constitutes. The major component in watermelon rind is citrulline that has a strong antioxidant effect which protect body from free-radical damage. Objective: This study was conducted to investigate the effect of microwave powers (150 W, 300 W & 450 W) and time intervals (1, 3 & 5 minutes) on total phenolic content (TPC) and total flavonoid content (TFC) and antioxidant characteristics i.e. DPPH and ferric reducing antioxidant potential (FRAP) of microwave assisted extracts of watermelon rind powder. Methods: The extracts collected after Microwave assisted extraction (MAE) of watermelon rind wereanalyzed for their antioxidant potential through different tests including total phenolic contents (TPC), total flavonoid content (TFC), DPPH assayand FRAP. Results: Microwave assisted extraction by using ethanol as a solvent at different microwave powers and various time intervals showed that total antioxidant potential was significantly higher at low microwave power such as TPC ranges obtained at 150W for 1, 3 & 5 minutes of time intervals show ranges (159.84, 160.04 & 169.71 mg GAE/100 g). While TFC ranges at 150W for time 1, 3 & 5 minutes were (21.31, 24.15 & 42.20 mg CEQ/100g) whereas DPPH ranges at 150W for time 1, 3 & 5 minutes were (53.14, 54.87 & 68.17 % ascorbic acid inhibition) and FRAP values at 150W for time 1, 3 & 5 minutes were (201.71, 221.50 & 326.43 mg FE/100g). While high microwave power 450W can result in disruption of some antioxidants at various time intervals. Conclusions: Watermelon rind is a rich source of many antioxidants andmicrowave assisted extraction technique should be implemented in the food and nutraceutical industries and microwave assisted extracts of watermelon rind should be utilize for the development of new functional food to combat many health related problems
Pakistan is an agriculture-based country and in agriculture sector, livestock is contributing lions‘ share. The livestock has great potential of increments in dairy products if the losses due to infectious diseases, lower management, and lack of recognition of this sector as industry may be reduced to minimum level. As foot-and-mouth disease (FMD) is a severe, clinically acute, contagious vesicular disease of cloven-hoofed animals including wild and domesticated ruminants. Foot-and-mouth disease virus (FMDV) types O, A and Asia I are endemic in Pakistan. The losses due to this disease only have reached to billions of rupees every year. In first part of the study, suspected FMD samples from 374 animals (buffaloes and cattle) were collected including mouth swabs, vesicles, and serum and tongue tissue from dead animals from field outbreaks occurred in three provinces of Pakistan (Punjab, Khyber Pakhtunkhwa and Balochistan) during 2005-2009. The samples were subjected to virus isolation, Ag-ELISA, non-structural protein (NSP) ELISA, real-time RT-PCR (rRT-PCR), reverse transcriptase PCR (RT-PCR), amplification of VP1, P1 and whole genome, sequencing of VP1, P1 and whole genome. The sequences were then used in phylogenetic analysis using MEGA 4.0, to assess the movement of virus. Thirty eight viruses were isolated on cell culture. Thirty six viruses were confirmed as FMDV serotype O and two as serotype A. The viral isolates were sequenced and that include FMDV type O (32 VP1, 2 whole genome and 2 P1 sequences) and type A (1 VP1 and 1 P1 sequences). Phylogenetic analysis revealed that all the isolates fall in PanAsia II lineage of a broader group of PanAsia lineage. The type A viral isolates were closely related to Iranian sequences of 2001, 2002 and 2003. The second part of the study was the development and evaluation of an ELISA system for the early detection of specific FMD IgM antibodies in the serum of infected, carrier and vaccinated animals. Experimental and field outbreak serum samples (n=3440) were used for this purpose. The experimental samples were from cattle and sheep experiments carried out at Institute for Animal Health (IAH), United Kingdom in the past and field outbreak serum samples were from UK 2001 and 2007 outbreaks of FMD. Naïve samples (n=236 cattle & 36 sheep) were tested to find out the frequency distribution. It was found that at a cutoff value of 0.3, 100% specificity can be achieved. Furthermore, the test also had the potential to detect the infected, vaccinated and carrier animals. It was concluded from the study that FMD was endemic in Pakistan and upon phylogenetic analysis it was revealed that same viruses with minor nucleotide changes were the cause of repeated outbreaks in the areas under study. To our knowledge, this is the first report on whole length genome sequence including S, L, and complete open reading frame of FMD viral isolate from Pakistan. Overall, the results of the IgM ELISA may correlate with the infection status and possibly the carrier status of cattle. Therefore, the IgM ELISA could be a tool to identify infected cattle and in particular probable carrier cattle in a vaccinated population. However, the IgM ELISA cannot, sensu strict, detect carrier animals, which is only possible by virus isolation. Further studies with vaccinated, non-infected cattle should be carried out to clarify whether the IgM ELISA can reliably detect infected cattle in a vaccinate population.