Chemical insecticides are widely used to control the insects, these insecticides are not environmentally healthy as they are not biodegradeable and hence are biomagnified. These insecticides are also not host specific; they also kill the beneficial insects. So there is a need to search for a control agent which should not harm the environment and also to human beings. Several different methods have been used in recent past to control the insects which include use of pheromones for trapping or disruption of mating behavior, insect growth regulators that interfere with larval development, parasitoids, fungi, viruses and bacteria, which debilitate or cause death in the infected insect. One of the most successful biological control organisms is a naturally occurring bacterial pathogen, Bacillus thuringiensis generally known as B.t. Formulations based on B.t. have been used for decades as biological insecticides for agriculture and forestry, as well as for vector control against mosquitoes and black flies. Interest in B.t. proteins has increased during the last two decades because of their unique qualities which are unmatched by any conventional insecticide. Of the 297 genes known to encode B.t. proteins, some share a high degree of homology, while others have diverse nucleotide sequences. Because of the interest in B.t., the list of new B.t. subspecies is growing as is the group of economically important target insects. B.t. produces crystal proteins during sporulation. These crystal proteins are of two types Cry and Cyt. Both of these types of proteins are different in their mode of action. Cytolytic proteins have an additional property of having cytolytic activity against different cells and also against mammalian erythrocytes. These proteins especially Cyt proteins are active against mosquito larvae. Recent studies reported the development of resistance in mosquitoes against Cry toxins. Researchers tried different methods to overcome this resistance and they found that when Cyt proteins are used in combination with Cry proteins they greatly reduced the resistance of mosquitoes against B.t. toxins. This indicated that Cyt proteins work synergistically with Cry proteins. In the present study, soil samples collected from different areas of Lahore, Kasur and Faisalabad. A total of 50 soil samples were collected, these soils were rich in organic manure. B.t. like bacteria were isolated from these soil samples using differential medium containing sodium acetate buffer. These isolates were then subjected to biochemical characterization by performing biochemical tests. The expected B.t. like isolates were screened for the presence of cyt genes. After confirmation of presence of cyt 2B gene, mosquitocidal activity of these isolates were checked by using B.t. spores and total B.t. cell proteins against 3rd instar larvae. From the bioassays, it was found that NB B.t.4 was found to be most toxic with LC50 value of 400±1.15 μg/ml and 68±0.46 μg/ml for its spores and total cell protein, respectively. After bioassays, six most toxic B.t. isolates were then selected for further study. Ribotyping of these isolates was done to amplifying 16S rRNA gene to identify these isolates. Protein profile of these isolates was checked to confirm the presence of 29 kDa protein band. Full length cyt 2B gene was amplified, cloned in pTZ57RT cloning vector, and pET22b vector was used for expression. IPTG induction of 1 mM was found good for expression ranging incubation time of 4-6 hours. Expressed protein was then purified by anion exchange chromatography. Bioassays were performed using recombinant organism (E. coli transformed with cyt 2B gene), expressed crude protein and purified protein. It was found that the purified protein was most toxic with LC50 Value of 50±1.68 μg/ml.
Human papilloma virus (HPV) related head and neck squamous cell cancer (HNSCC) has varying etiology, genetic as well as environmental factors involved and differential clinicicopathological features. HNSCC came in the limelight recently due to increased incidence rate and insufficient diagnostic methods. This review will comprehensively focus on the characteristics of HPV associated HNSCC. It will provide an updated review of our understanding of HPV role in Oral squamous cell carcinoma (OSCC) known to date. Curruntly, three vaccines are available (Gardasil, Gardasil 9 and Cervarix). These vaccines prevent infections with HPV types 16 and 18 HPV-16 is most common type associated with HNSCC. HPV related HNSCC has better prognosis, does not mutate but inactivatestumor suppressor genes and therefore has comparatively better treatment options. However, there is still a need to improve our methods of sampling, HPV molecular assay and type of specimen to be used.
Maize germs from six promising locally grown hybrids, namely Poineer-32-F-10 (P-1), Poineer-32-B-33 (P-2), Monsanto-6142 (M-1), Monsanto-6525 (M-2), Rafhan- 2331 (R-1) and commercial were characterized for proximate composition, selected minerals, amino acid & fatty acid profiles and tocopherols. Based on the germ recovery and compositional analysis, one best hybrid germ, Pioneer 32-F-10 (P-1), was selected for further analysis and product development. Refined maize germ oil (MGO) from selected hybrid, was analyzed for various physical and chemical characteristics. The MGO was evaluated for cake preparation through blending with normal shortening at different levels. Moreover, defatted maize germ (DMG) meal was subjected to biological and safety evaluation using Sprague Dawley rats. Later, DMG flour was blended with wheat flour in different combinations; evaluated for functional properties, textural analysis (SMS Texture Analyzer) and Farinographic behavior. Flour blends were further used to develop value-added products like bread and cookies. Germ recovery from the experimental hybrids ranged from 6.31-7.68%, while the highest germ yield was observed in P-1 hybrid. Germ samples were found to be nutrient dense; crude protein 16.34-20.96%, crude fat 32.10-38.80%, crude fiber 2.63-4.79%, ash 3.08-4.94%, α-tocopherol 4.63-9.68 mg/100g and γ-tocopherol 29.52-35.51 mg/100g. The germ samples were rich in polyunsaturated fatty acids (46.74- 58.00 g/100g) and essential amino acids (33.30-39.04 g/100g). Additionally, in germ samples, minerals like P (1.06-1.79 g/100g), K (1.19-1.64 g/100g), Mg (0.43- 0.78 g/100g) and Fe (9.08-14.46 mg/100g) were in substantial amount. MGO from selected germ (P-1) was successfully incorporated in cake recipe with high sensory quality. Defatted maize germ (DMG) meal was found to be considerable source of protein (32.1%), dietary fiber (31.87%) and allied minerals. In the experimental rats, in-vivo protein quality of DMG flour was: 87.10±0.78% true digestibility, 76.70±1.25% net protein utilization, 88.06±o.67% biological value, 5.12±0.21 net protein ratio and 2.15±0.03 protein efficiency ratio were significantly higher than that of wheat-based diet and comparable with casein. Favorable impact of DMG flour on serum biochemical profile was observed; cholesterol, triglyceride, LDL and glucose levels decreased up to 6.80, 12.45, 16.19 and 6.50 %, respectively. DMG flour resulted in improvement of functional properties of DMG flour-wheat flour blends with special reference to improved water & oil absorption capacity, gelling and emulsion properties; revealed its worth in food preparations. Protein and fiber enriched cookies and breads were prepared up to 15% wheat flour substitution with DMG flour. Consumer response to purchase defatted maize germ flour fortified cookies was very positive e.g. 64% of the respondents said that they would prefer to purchase.