In vitro studies using curcumin and curcumin analogues as candidate mitochondria-targeting anticancer agents affecting colon cancer cells

Curcumin is one of the major curcuminoids produced by the ginger family Zingiberaceae. These curcuminoids possess pharmacological properties that include anticancer activities. We have evaluated some synthetic curcumin analogues that have shown potential as anticancer drugs. These antineoplastic agents bearing the 1,5-diaryl-3-oxo-1,4-pentadienyl pharmacophore are electrophiles which are designed to preferentially react with sulfhydryl groups present in proteins as opposed to amino and hydroxyl groups present in DNA. In previous pilot studies, three derivatives examined in this thesis showed inhibition towards human cancer cell lines such as Molt 4/C8 and CEM T-lymphocytes. In this thesis work, I determined the cytotoxicity of these derivatives and curcumin towards human colon cancer (HCT-116) cells and also normal colon epithelial (CRL-1790) cells, and examined the possible mechanism(s) involved. I hypothesized that they act via induction of reactive oxygen species (ROS) which elicit a transient surge of mitochondrial ROS generation and a phenomenon known as ROS-induced ROS release (RIRR), along with the mitochondrial permeability transition (MPT) and mitochondrion –dependent apoptosis. I asked whether these agents react with some of the key protein thiols in the mitochondria whose oxidation/alkylation results in mitochondrion - dependent apoptosis. NC-2109 and NC-2346 were found to be potent cytotoxic agents based on their GI50 values of 0.87 ± 0.38 μM and 0.90 ± 0.22 μM, respectively, and were more potent than the anticancer drug 5-fluorouracil (GI50 = 5.47 ± 0.55 μM) and curcumin (GI50 = 3.50 ± 0.36 μM). However NC-2109 was found to have a better selectivity towards cancer cells over normal cells (a selectivity index of 18.81 versus 5-FU, curcumin and NC-2346 which had selectivity indices of 1.87, 16.75 and 4.61, respectively). In the investigations of the mechanisms involved, both curcumin and curcumin analogues were able to induce mitochondrial ROS production. Moreover, curcumin and its synthetic counterparts showed a biphasic ROS profile which is most characteristic of RIRR. Treatment with these agents also led to the disruption of the mitochondrial membrane potential, suggesting oxidation of protein thiols and the opening of the mitochondrial permeability transition pore which is an important step to initiate mitochondria-directed apoptosis. This possibility was confirmed based on GSSG/GSH ratios, since curcumin, NC-2346 and NC-2109 all produced a higher GSSG/GSH ratio than the controls. In addition to their ability to depolarize the mitochondrial membrane in HCT-116 cells, that these molecules acted via the mitochondrial pathway were further authenticated based on their ability to induce mitochondrial swelling in rat liver mitochondria. In another part of this thesis I evaluated the involvement of the critical thiol protein adenine nucleotide translocase (ANT), a bifunctional protein that plays a central role in mitochondrial apoptosis. ANT has four different isoforms; ANT1 and ANT3 are proapoptotic, while ANT2 and ANT4 are antiapoptotic and are overexpressed in cancer states. A combination approach using ANT2 siRNA however did not conclusively show whether these agents acted synergistically with ANT2 knockdown to potentiate mitochondria-mediated cell death. An alternative combination approach was the use of carboxyatractyloside (CAT) which binds to and retains ANT in its ‘c’ conformation, exposing thiols and potentially driving a cell towards programmed cell death. The presence of CAT enhanced the ability of curcumin and its synthetic analogs to collapse the mitochondrial membrane potential, an important step in mitochondrial-mediated apoptosis. In conclusion, curcumin and the curcumin analogue NC-2109 were found to be cytotoxic in vitro, towards HCT-116 cells and also showed good selectivity. In addition, these two molecules were found to be ROS inducers, and coincidentally oxidized cellular thiols and caused depolarization of the mitochondrial membrane potential. The results support a mechanism of mitochondrial-mediated cell death upon MPT pore formation (mitochondrial swelling), perhaps involving ANT2. This conclusion was further supported by the potentiation of cell death in the presence of the ANT2 inhibitor, CAT

The Relationship Between Employee Competencies And Employee Performance In A Government-Linked Company (GLC)

This study is aimed at investigating the relationship between employee competencies and employee performance in a government-linked company (GLC). This research as done by using quantitative method where the data were collected by using questionnaires as the instrument. The questionnaires were in the form of Google Forms online due to the pandemic situation and the data was collected from 460 samples in one of the government-linked company in Bintulu, Sarawak. The data were then analysed by using both descriptive and inferential statistics. Besides, Pearson Correlation test procedure was used to measure the relationships between the independent variables and the dependent variables. The finding have shown that there is a significant relationship between the employee competencies and the employee performance. It was also found that …. is the most dominant factor affecting employee performance. As such, the researcher has recommended a focus on the study that may guide the organizations and future researches

SPECTRO-ANALYTICAL, COMPUTATIONAL AND BIOLOGICAL STUDIES ON 4-PYRIDINE CARBOXALDEHYDE-3-HYDROXY-5-(HYDROXY METHYL)-2-METHYL HYDRAZONE HYDROCHLORIDE AND ITS CU (II) COMPLEXCA

Objective: The title compound 4-pyridine carboxaldehyde 3-hydroxy-5-(hydroxy methyl)-2-methyl hydrazone (PCHHMMH) hydrochloride an analogue of Pyridoxal isonicotinoyl hydrazone PIH, is an iron chelator. The PCHHMMH has potential donor sites suitable for metal ion binding, the study on structural aspects of the compound and its copper complex are explored. With a view to understand biological importance of title compounds, antimicrobial and cytotoxic studies were planned. Methods: In the present study the spectroanalytical techniques employed were pH-metry, spectrophotometry, IR, 1H & 13C-NMR, UV-Vis, ESR, Magnetic measurements, TGA and SEM. The computational method employed is HyperChem 7.5 software. The antimicrobial studies were carried out by agar disc diffusion method for antibacterial studies against Gram positive and Gram negative bacteria. The cytotoxic potential was measured by Sulforhodamine B (SRB) method against selected tumor cells. Results: The equilibrium studies by employing pH-metric method inferred the dissociation of two protons in it. Further titration in presence of Cu (II) ion, it is confirmed the release of two protons from title compound and formation of corresponding complex. The orientations of frontier orbitals for molecular and ionized forms of compound were computed to understand the electronic properties. The Cu (II) PCHHMMH complex was characterized by spectroanalytical methods and screened for, antimicrobial and cytotoxic activities. Conclusion: As the structural features are important to understand the chemical behavior of metal complexes, in the present study copper complex was synthesized and characterized by employing various spectro-analytical tools viz; IR, 1H & 13C-NMR, UV-Vis, ESR, Magnetic measurements, TGA and SEM. Further the antimicrobial and cytotoxic activities were evaluated and correlated with computed QSAR data

Identification of functional and diverse circulating cancer‐associated fibroblasts in metastatic castration‐naïve prostate cancer patients

In prostate cancer (PCa), cancer-associated fibroblasts (CAFs) promote tumor progression, drug resistance, and metastasis. Although circulating tumor cells are studied as prognostic and diagnostic markers, little is known about other circulating cells and their association with PCa metastasis. Here, we explored the presence of circulating CAFs (cCAFs) in metastatic castration-naïve prostate cancer (mCNPC) patients. cCAFs were stained with fibroblast activation protein (FAP), epithelial cell adhesion molecule (EpCAM), and receptor-type tyrosine-protein phosphatase C (CD45), then FAP+EpCAM− cCAFs were enumerated and sorted using fluorescence-activated cell sorting. FAP+EpCAM− cCAFs ranged from 60 to 776 (389 mean ± 229 SD) per 2 × 108 mononuclear cells, whereas, in healthy donors, FAP+ EpCAM− cCAFs ranged from 0 to 71 (28 mean ± 22 SD). The mCNPC-derived cCAFs showed positivity for vimentin and intracellular collagen-I. They were viable and functional after sorting, as confirmed by single-cell collagen-I secretion after 48 h of culturing. Two cCAF subpopulations, FAP+CD45− and FAP+CD45+, were identified, both expressing collagen-I and vimentin, but with distinctly different morphologies. Collectively, this study demonstrates the presence of functional and viable circulating CAFs in mCNPC patients, suggesting the role of these cells in prostate cancer

Flow-based immunomagnetic enrichment of circulating tumor cells from diagnostic leukapheresis product

The clinical utility of circulating tumor cells (CTCs) is hampered by the low number of cells detected. Diagnostic leukapheresis (DLA) offers a solution but, due to the observed non-specific binding and clumping, processing of DLA samples using the CellSearch system only allows for the processing of aliquots consisting of ~ 2% of the total DLA sample per test. Here, we introduce a flow enrichment target capture Halbach-array (FETCH)-based separation method in combination with a DNase preprocessing step to capture CTCs from larger fractions of DLA products without clumping. To evaluate the FETCH method, we processed peripheral blood samples from 19 metastatic castration-naïve prostate cancer (mCNPC) patients with CellSearch, and processed 2% aliquots of leukapheresis samples from the same patients with CellSearch as well as FETCH with or without DNase preprocessing. Using 2% aliquots from six patients, the use of FETCH with fewer immunomagnetic epithelial cellular adhesion molecule (EpCAM) conjugated ferrofluids was tested, whereas 20% aliquots from four patients were used to evaluate the processing of 10-fold larger DLA samples using FETCH. Results show that the cell clumping normally seen after immunomagnetic enrichment of DLA material was greatly reduced with the use of DNase pretreatment, while the number of CTCs detected was not affected. The number of CTCs detected in 2% aliquots of DLA using FETCH was unchanged compared to CellSearch and did not decrease when using down to 10% of the volume of immunomagnetic anti-EpCAM ferrofluids normally used in a CellSearch test, whereas the number of co-enriched white blood cells reduced a median 3.2-fold. Processing of a 20% aliquot of DLA with FETCH resulted in a 14-fold increase in CTCs compared to the processing of 2% aliquots of DLA using CellSearch and a total 42-fold median increase in CTCs compared to peripheral-blood CellSearch.</p

Covid-19 and Oral Health

The COVID-19 pandemic has undoubtedly changed many aspects of health and healthcare on a global scale. We have heard about the impact that this pandemic has had on mental health status and general well- being; however, a topic that has been overlooked is the relationship between COVID-19 and oral health. Given that transmission of SARS-CoV-2 occurs through contact with respiratory droplets, the oral cavity can play a significant role in the spread of the disease. Thus, analyzing this relationship further may provide insight into managing COVID-19 outcomes

An Algorithm for Garbage Collection in Multicomputer Systems

There is widespread interest in multicomputer parallelism. Functional languages with their inherent parallelism can form a basis for programming these machines. These languages dynamically allocate memory, objects are created when required and eventually objects may lose links with active objects and become unreachable garbage. The process of recovering these inactive objects is called garbage collection. Garbage collection in a multicomputer system has to manage objects in physically separated memories. This introduces consistency and synchronization problems with the shared data. The development of an algorithm for garbage collection in a multicomputer system is reported. A description of the algorithm is presented. The design of the simulator and the simulation experiments are presented. The application of Petri nets to the modelling of this algorithm is discussed. Verification of some properties of this algorithm using the invariants of the Petri net model are presented

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Not AvailableA study was conducted to screen eight sunflower restorer lines for tolerance to drought under field conditions based on yield characteristics. Moisture stress was imposed in stress plots from 45 DAS to harvest. Whereas, control plots were irrigated at 10 days intervals throughout the crop growth period. Results revealed that water stress showed repressing effect on yield related attributes like capitulum dry weight by 36%, total number of seeds per capitulum by 25%, seed yield per plant by 34% compared to control. Significant difference among R-lines was observed for head weight, test weight, harvest index and oil content. R-lines RGP 33-P5, RGP 50-P1, RGP 61-P1and RGP 61-P2 showed high HI under water stress conditions. Most of the R-lines have increased oil content under stress. RGP 50-P1, RGP 60-P2 and RGP 61-P1 have decreased oil content under stress. Based on Stress Tolerance Index and seed yield under stress, RGP 21-P6 and RGP 61-P1were identified as tolerant to water stress out of eight R-lines studied. These lines also showed superior morphological, physiological traits. RGP 32-P1 and RGP 33-P5 were found to be more sensitive.Not Availabl

Socio-ecological determinants of malaria transmission risk among population residing in an endemic area of southern province of Karnataka, India

Background: India accounts for 79% of global malaria burden. With progress towards malaria elimination, assessment of true disease burden and determinants of its transmission risk is critical. The present study aimed to estimate the burden of malaria and identify socioecological factors determining the disease prevalence. Methods: A community-based cross-sectional study was carried out among 839 households covering 2910 individuals residing in Udupi taluk. A representative sample from urban, rural tribal and migrant communities were recruited and a questionnaire was used to collect information on socio-demographic, housing &amp; behavioral factors. Data was collected using Epi-info mobile application version 7.2.4 and analyzed using SPSS version 16.0. Results: Among the population covered a total of 15 participants had a history of malaria in the past. Advancing age (AOR = 3.60; 95% CI: 2.14–6.04), illiteracy (AOR = 2.28; 95% CI:1.35–3.85), (AOR = 1.96; 95% CI:1.24–3.11) and rural (AOR = 1.96; 95% CI:1.39–2.74), tribal (AOR = 2.38; 95% CI:1.49–3.79), migrant (AOR = 4.28; 95% CI:2.30–7.98) localities were found to be independent predictors of disease awareness. With respect to malaria prevention practices, households from middle (AOR = 0.73; 95% CI: 0.49–1.09) &amp; low SES (AOR = 0.30; 95% CI: 0.19–0.46) and those from migrant localities (AOR = 0.66; 95% CI: 0.38–1.14) were less likely to have poor practices. Conclusion: Age, education, socioeconomic status, and locality were found to be significantly associated with knowledge and practice towards malaria prevention. Creating awareness regarding malaria and its prevention with enhanced community participation in control activities jointly with local authority is further emphasized

AN EFFICIENT CASHLESS TOLL COLLECTION SYSTEM USING RFID TAG

In order to cut down the manual work at toll plazas, a Radio Frequency Identification (RFID) based concept is portage for the entire process within a second. The system uses passive tag which will have the particular number mapping the owner information; this useful detail is stored in congruence to that particular number.&nbsp;ATCS known as "Automatic Toll Collection System" is a technology used to reduce the amount of time spent in toll queue, provide passenger leisure of plaza payment, illegal entry and to detect stolen vehicle. A reader antenna is normally assembled above the lane. It picks up the transponder's signal. The distinct number is then deliver to the globalize server through local area network, forth with plazas-location and lane information.The system then matches that distinct number and specific reduction takes place through the e-wallet reference to correlated RFID tag that reside to the owners account. In case a robbed automobile passes through the plaza collection center, it is encounter and the proclamation is sent to the Police station of that area