Serum circulating cell free DNA as potential diagnostic and prognostic biomarker in non small cell lung cancer

BackgroundNon-small cell lung cancer (NSCLC) is leading cause of cancer related death and the survival rate for patients with NSCLC remain poor so early diagnosis of NSCLC represents the best opportunity for cure. Cell-free DNA (cf-DNA) is extracellular nucleic acids found in cell-free plasma/serum of humans, given the recent approval of a liquid biopsy in lung cancer, the use of circulating tumor DNA as a novel non-invasive diagnostic and prognostic biomarker is promising.ObjectivesStudying whether the concentrations of circulating Cell Free DNA in serum can be used as a diagnostic and prognostic biomarker for NSCLC patients.MethodThis study was carried out on 140 subjects included 60 patients with non small cell lung cancer,40 patients with Chronic Obstructive Pulmonary Disease (COPD) and 40 healthy controls. Quantitative analysis of serum circulating cf-DNA was done b y AlU-based quantitative real time PCR. Serum level of CEA was measured by ELISA.ResultsNSCLC patients demonstrated significantly higher values of each of ALU 215, ALU 247, and DNA integrity than both COPD patients and controls. On ROC curve analysis, the total accuracy of ALU 247, ALU 115, DNA integrity (92.1%, 83.6%, 56.4%) at cutoff points (325, 565 & 0.48) respectively. On combining both DNA integrity and CEA, improved sensitivity to 93.3% was noted. For NSCLC patients, ALU 115 & ALU 247 increased significantly with more advanced stage and highest level was noticed in metastatic patients. Regarding survival there was better overall survival among patients with low DNA integrity.ConclusionSerum cf-DNA concentrations and integrity index may be valuable tool in early diagnosis of NSCLC and prediction of prognosis of those patients.     

Floristic inventory of wild plants of Peshawar university campus

Floristic inventory of wild plants comprised of total 129 plant species belonging to 42 families and 101 genera. Location wise UAP contributed greater amount of plant species (53%), followed by PFI (29%), UoP (15%) and Islamia College (3%). Majority of the plants were annual herbs (58%), followed by perennial herbs (29%), trees (10%) and shrubs (3%), respectively. The leading families included Poaceae, contributed 18% of all the plant species. Habitat wise, 44% of them grew in grassy plains, abandoned lands and mismanaged lawns, followed by roadsides (34%), undulating grassy plains (15%) and stony grounds with uneven topography along with stream banks and drainage ditches (7%). Still some of plant species were used as ornamentals, while some shrubs were use in fencing around fields; others were utilized as green manure, for fishing, sheltering and religious purposes. Exploring and identifying the wild flora of the campus and compiling as a handbook would help to monitor new plants introductions particularly of noxious invasive weeds in the campuses in future.     

Inhibition of gut proteases and development of dengue vector, Aedes aegypti by Allium sativum protease inhibitor

The paper describes the bio efficacy of a protease inhibitor; isolated from Allium sativum ‘garlic’ (ASPI); against Aedes aegypti mosquito, a well-known transmitter of dengue and Chikungunya. The purification of protease inhibitor from Allium sativum ‘garlic’ (ASPI) was carried out by ammonium sulfate precipitation followed by Fast Protein Liquid Chromatography using akta DEAE-Cellulose column. The protein fraction demonstrating trypsin inhibitory activity was further evaluated for its insecticidal activity using gut protease inhibition assay and larvicidal assay. ASPI is an inhibitor of porcine trypsin (IC₅₀ of 650.726?μg/mL) and has molecular weight of ~15?kDa determined by SDS PAGE similar to other inhibitors of the Kunitz-type family (14–26?kDa). ASPI demonstrated 50% reduced activity of Ae. aegypti midgut proteases and showed a dose-dependent acute toxicity on Ae. aegypti 3rd instars exhibiting LC₅₀ value of ~50.827?μg/mL. After ten days of larval exposure ASPI resulted in a 24-h delay of larval development and ~72% mortality at 61.5?μg/mL. These results suggest that ASPI may serve as potent insecticidal agent and hence opens a new gateway in the field of phyto-remediation.     

Earliness index deters <Emphasis Type="Italic">Pectinophora gossypiella</Emphasis> incidence on advanced cultivars of <Emphasis Type="Italic">Bt</Emphasis> cotton

Pink bollworm (Pectinophora gossypiella) is recognized as an important pest of cotton and can damage flowers and bolls of both Bt and non-Bt cultivars. Cry-1Ac in Bt cultivars is considered very effective in controlling lepidopterous larvae; therefore, the present study was carried out to investigate the impact of Cry1-Ac and the earliness index on the natural incidence of P. gossypiella at the Cotton Research Institute, Faisalabad. During 2015–2016, ten cultivars were used to determine the incidence of pink bollworm infestation. The experiment was repeated for 2 years. During the next year, Cry1-Ac and earliness traits of selected cultivars were also observed to determine their impact on pink bollworm. Correlation coefficient results regarding days to first flower (r value = 0.66) as well as the earliness index (r value = ? 0.62) exhibited a strong association with pink bollworm, but Cry1-Ac had a weak association (r value = ? 0.058) with pink bollworm. The coefficient of determination (R ²) explained that variability of pink bollworm due to Cry1-Ac, the earliness index, and days to first flower was 18.0, 38.5, and 43.5%, respectively. Principal component analysis results showed that the first two PCs expressed 87% of the total variability. Clusters made on the basis of the studied parameters revealed that clusters 2 and 3 comprised the cotton cultivars possessing earliness traits compared with cluster 1. Therefore, it can be concluded that the earliness index in cotton is an important component for the sustainable management of pink bollworm infestation, the need for which is endless to evade the pink bollworm problem in the era of climate change.     

Systematic relationships within the Microtus arvalis (Rodentia: Cricetidae) group in Iran,inferred from cytogenetic analyses

The distribution of C-heterochromatin and nucleolar organizer regions (NORs) was studied in three species of voles of the Microtus arvalis group in Iran: M. mystacinus, M. kermanensis, and M. transcaspicus. The C-banding pattern and NORs distribution were similar in M. mystacinus and M. kermanensis suggesting taxonomic proximity of these two species. At the same time, the karyotypes of M. mystacinus from Iran were different in C-banding pattern from the complements of conspecific 54-chromosome voles from Europe and other regions of Asia. The most distinct difference was in size of the distal C-positive block of heterochromatin on the X chromosome. In this respect M. mystacinus from Iran and M. kermanensis resembled M. transcaspicus. Small size of the distal C-positive heterochromatic block may be ancestral whereas larger size is derived. The X chromosome of M. transcaspicus can be derived from that of M. mystacinus and M. kermanensis by a large inversion or centromeric shift.     

Current status and future possibilities of molecular genetics techniques in <Emphasis Type="Italic">Brassica napus</Emphasis>

As PCR methods have improved over the last 15 years, there has been an upsurge in the number of new DNA marker tools, which has allowed the generation of high-density molecular maps for all the key Brassica crop types. Biotechnology and molecular plant breeding have emerged as a significant tool for molecular understanding that led to a significant crop improvement in the Brassica napus species. Brassica napus possess a very complicated polyploidy-based genomics. The quantitative trait locus (QTL) is not sufficient to develop effective markers for trait introgression. In the coming years, the molecular marker techniques will be more effective to determine the whole genome impairing desired traits. Available genetic markers using the single-nucleotide sequence (SNP) technique and high-throughput sequencing are effective in determining the maps and genome polymorphisms amongst candidate genes and allele interactions. High-throughput sequencing and gene mapping techniques are involved in discovering new alleles and gene pairs, serving as a bridge between the gene map and genome evaluation. The decreasing cost for DNA sequencing will help in discovering full genome sequences with less resources and time. This review describes (1) the current use of integrated approaches, such as molecular marker technologies, to determine genome arrangements and interspecific outcomes combined with cost-effective genomes to increase the efficiency in prognostic breeding efforts. (2) It also focused on functional genomics, proteomics and field-based breeding practices to achieve insight into the genetics underlying both simple and complex traits in canola.     

Effects of Modafinil on Clonic Seizure Threshold Induced by Pentylenetetrazole in Mice: Involvement of Glutamate,Nitric oxide,GABA, and Serotonin Pathways

Epilepsy is the third most common chronic brain disorder. Modafinil is an awakening agent approved for narcolepsy. In addition to its clinical uses some reports revealed that modafinil was associated with some alterations in seizure threshold. The purpose of this study was to clarify the effect of acute administration of modafinil in clonic seizure threshold (CST) induced by pentylenetetrazole in mice and the involvement of glutamate, nitric oxide, gamma amino butyric acid (GABA), and serotonin systems in this feature. Modafinil at 80 and 150 mg/kg showed anti- and pro-convulsant effects respectively and expressed maximum anti- and pro-convulsant activities at 30 min after injection. Both modulatory effects were blunted by pretreatment of l-NAME [nonspecific nitric oxide synthase (NOS) inhibitor; 10 mg/kg, i.p.], 7-nitroindazole (a neuronal NOS inhibitor; 40 mg/kg, i.p.), and aminoguanidine (an inducible NOS inhibitor; 50 mg/kg, i.p.). Injection of the NOS precursor l-arginine (60 mg/kg, i.p.) before modafinil did not change the anti-convulsant effect, while thoroughly reversed the pro-convulsant one. Our experiments displayed that administration of diazepam (a GABA_A receptor agonist; 0.02 mg/kg, i.p.) and MK-801 (a NMDA receptor antagonist; 0.05 mg/kg, i.p.) before different doses of modafinil significantly increased CST. Finally, pretreatment of citalopram (a selective serotonin reuptake inhibitor; 0.1 mg/kg, i.p.) did not modify the convulsant activities of modafinil. Therefore, nitric oxide system may mediate anti-convulsant activity, while glutamate, nitric oxide, and GABA pathways may involve in pro-convulsant property. Serotonin receptors have no role on convulsant effects of modafinil.     

Deletion of RB exons 24 and 25 causes low-penetrance retinoblastoma.

A deletion in the tumor-suppressor gene, RB, discovered by quantitative multiplex PCR, shows low penetrance (LP), since only 39% of eyes at risk in this family develop retinoblastoma. The 4-kb deletion spanning exons 24 and 25 (delta24-25) is the largest ever observed in an LP retinoblastoma family. Unlike the usual RB mutations, which cause retinoblastoma in 95% of at-risk eyes and yield no detectable protein, the delta24-25 allele transcribed a message splicing exon 23 to exon 26, resulting in a detectable protein (pRBdelta24-25) that lacks 58 amino acids from the C-terminal domain, proving that this domain is essential for suppression of retinoblastoma. Two functions were partially impaired by delta24-25-nuclear localization and repression of E2F-consistent with the idea that LP mutations generate "weak alleles" by reducing but not eliminating essential activities. However, delta24-25 ablated interaction of pRB with MDM2. Since a homozygous LP allele is considered nontumorigenic, the pRB/MDM2 interaction may be semi- or nonessential for suppressing retinoblastoma. Alternatively, some homozygous LP alleles may not cause tumorigenesis because an additional event is required (the "three-hit hypothesis"), or the resulting imbalance in pRB function may cause apoptosis (the "death allele hypothesis"). pRBdelta24-25 was also completely defective in suppressing growth of Saos-2 osteosarcoma cells. Targeting pRBdelta24-25 to the nucleus did not improve Saos-2 growth suppression, suggesting that C-terminal domain functions other than nuclear localization are essential for blocking proliferation in these cells. Since delta24-25 behaves like a null allele in these cells but like an LP allele in the retina, pRB may use different mechanisms to control growth in different cell types.