Abstract: Yeast cells are generally used as a model system of eukaryotes due to their complex genetic structure, rapid growth ability in optimum conditions, easy replication and well-defined genetic system properties. Thus, yeast cells increased the knowledge of the principal pathways in humans. During fermentation, carbohydrates (hexoses and pentoses) degrade into some toxic by-products such as 5-hydroxymethylfurfural (5-HMF or HMF) and furfural. HMF influences the ethanol yield, and ethanol productivity; it interferes with microbial growth and is considered as a potent inhibitor of bioethanol production. In this study, yeast single cell behavior under HMF application was monitored by using a continuous flow single phase microfluidic platform. Microfluidic device in operation is fabricated by hot embossing and thermo-compression techniques from cyclo-olefin polymer (COP). COP is biocompatible, transparent and rigid material and it is suitable for observing fluorescence of cells considering its low auto-fluorescence characteristic. The response of yeast cells was recorded through Red Fluorescent Protein (RFP) tagged Nop56 gene product, which is an essential evolutionary-conserved nucleolar protein, and also a member of the box C/D snoRNP complexes. With the application of HMF, yeast cell proliferation continued but HMF slowed down the cell growth, and after HMF treatment the cell proliferation stopped. By the addition of fresh nutrient medium, the yeast cells recovered after 6 hours of HMF exposure. Thus, HMF application suppresses normal functioning of cell cycle but it does not cause cells to die. The monitoring of Nop56 expression phases of the individual cells shed light on the protein and ribosome synthesis cycles along with their link to growth. Further computational study revealed that the mechanisms underlying the inhibitory or inductive effects of HMF on growth are enriched in functional categories of protein degradation, protein processing, DNA repair and multidrug resistance. The present microfluidic device can successfully be used for studying the effects of inhibitory agents on growth by single cell tracking, thus capturing cell to cell variations. By metabolic engineering techniques, engineered strains can be developed, and the metabolic network of the microorganism can thus be manipulated such that chemical overproduction of target metabolite is achieved along with the maximum growth/biomass yield.
Abstract: 5-FU is a chemotherapeutic agent that has been used in colorectal cancer (CRC) treatment. However, it is usually associated with the acquired resistance, which decreases the therapeutic effects of 5-FU. miR-200c is involved in chemotherapeutic drug resistance, but its mechanism is not fully understood. In this study, the effect of inhibition of miR-200c in sensitivity of HCT-116 CRC cells to 5-FU was evaluated. HCT-116 cells were transfected with LNA-anti- miR-200c for 48 h. mRNA expression of miR-200c was evaluated using quantitative real- time PCR. The protein expression of phosphatase and tensin homolog (PTEN) and E-cadherin were analyzed by western blotting. Annexin V and propidium iodide staining assay were applied for apoptosis detection. The caspase-3 activation was evaluated by an enzymatic assay. The results showed LNA-anti-miR-200c inhibited the expression of PTEN and E-cadherin protein, apoptosis and activation of caspase 3 compared with control cells. In conclusion, these results suggest that miR-200c as a prognostic marker can overcome to 5-FU chemoresistance in CRC.
Abstract: The viewpoint towards the use of drugs or vaccines against avian parasitic diseases is one of the most striking challenges in avian medical parasitology. This includes many difficulties associated with drug resistance and in developing prophylactic vaccines. In many instances, the potential success of a vaccination in controlling parasitic diseases in poultry is well-documented. However, some medical, technical and financial limitations are still paramount. On the other hand, chemotherapy is not very well-recommended due to a number of medical limitations. But in the absence of an effective vaccine, drugs are used against parasitic diseases. This paper sheds light on some the advantages and disadvantages of using vaccination and drugs in controlling parasitic diseases in poultry species. The usage of chemotherapeutic drugs is discussed with some examples. Then, more light will be shed on using vaccines as a potentially effective and promising control tool.
Abstract: Substandard and counterfeit antimalarials is a major problem in malaria endemic areas. The availability of counterfeit/ substandard medicines is not only decreasing the efficacy in patients, but it is also one of the contributing factors for developing antimalarial drug resistance. Owing to this, a pilot study was conducted to survey quality of drugs collected from different malaria endemic areas of India. Artesunate+Sulphadoxine-Pyrimethamine (AS+SP), Artemether-Lumefantrine (AL), Chloroquine (CQ) tablets were randomly picked from public health facilities in selected states of India. The quality of antimalarial drugs from these areas was assessed by using Global Pharma Health Fund Minilab test kit. This includes physical/visual inspection and disintegration test. Thin-layer chromatography (TLC) was carried out for semi-quantitative assessment of active pharmaceutical ingredients. A total of 45 brands, out of which 21 were for CQ, 14 for AL and 10 for AS+SP were tested from Uttar Pradesh (U.P.), Mizoram, Meghalaya and Gujrat states. One out of 45 samples showed variable disintegration and retension factor. The variable disintegration and retention factor which would have been due to substandard quality or other factors including storage. However, HPLC analysis confirms standard active pharmaceutical ingredient, but may be due to humid temperature and moisture in storage may account for the observed result.
Abstract: Urinary Tract Infections are considered as one of the
most common bacterial infections with an estimated annual global
incidence of 150 million. Antimicrobial drug resistance is one of the
major threats due to wide spread usage of uncontrolled antibiotics. In
this study, a total number of 9149 urine samples were collected from
R.H Patiala and processed in the Department of Microbiology G. M.
C Patiala (January 2013 to December 2013). Urine samples were
inoculated on MacConkey’s and blood agar plates and incubated at
370C for 24 hrs. The organisms were identified by colony characters,
Gram’s staining, and biochemical reactions. Antimicrobial
susceptibility of the isolates was determined against various
antimicrobial agents (Hi – Media Mumbai India) by Kirby Bauer
DISK diffusion method on Muller Hinton agar plates. Maximum patients were in the age group of 21-30 yrs followed by
31-40 yrs. Males (34%) are less prone to urinary tract infections than
females (66%). Culture was positive in 25% of the samples.
Escherichia coli was the most common isolate 60.3% followed by
Klebsiella pneumoniae 13.5%, Proteus spp. 9% and Staphylococcus
aureus 7.6%. Most of the urinary isolates were sensitive to,
carbepenems, Aztreonam, Amikacin, and Piperacillin + Tazobactum.
All the isolates showed a good sensitivity towards Nitrofurantoin
(82%). ESBL production was found to be 70.6% in Escherichia coli
and 29.4% in Klebsiella pneumonia. Susceptibility of ESBL
producers to Imipenem, Nitrofurantoin and Amikacin were found to
be 100%, 76%, and 75% respectively. Uropathogens are increasingly
showing resistance to many antibiotics making empiric management
of outpatient UTIs challenging. Ampicillin, Cotrimoxazole and
Ciprofloxacin should not be used in empiric treatment. Nitrofurantoin
could be used in lower urinary tract infection. Knowledge of
uropathogens and their antimicrobial susceptibility pattern in a
geographical region will help in appropriate and judicious antibiotic
usage in a health care setup.
Abstract: With drug resistance becoming widespread in
Plasmodium falciparum infections, the development of the alternative
drugs is the desired strategy for prevention and cure of malaria. Three
drug targets were selected to screen promising drug molecules from
the GSK library of 13469 molecules. Using an in silico structure-based
drug designing approach, the differences in binding energies of
the substrate and inhibitor were exploited between target sites of
parasite and human to design a drug molecule against Plasmodium.
The docking studies have shown several promising molecules from
GSK library with more effective binding as compared to the already
known inhibitors for the drug targets. Though stronger interaction has
been shown by several molecules as compared to the reference, few
molecules have shown the potential as drug candidates though in
vitro studies are required to validate the results. In case of
thymidylate synthase-dihydrofolatereductase (TS-DHFR), three
compounds have shown promise for future studies as potential drugs.
Abstract: Second line antiretroviral therapy (ART) regimen is
used when patients fail their first line regimen. There are many
factors such as non-adherence, drug resistance as well as virological
and immunological failure that lead to second line highly active
antiretroviral therapy (HAART) regimen treatment failure. This study
was aimed at determining predictor factors to treatment failure with
second line HAART and analyzing median survival time.
An observational, retrospective study was conducted in Sungai
Buloh Hospital (HSB) to assess current status of HIV patients treated
with second line HAART regimen. Convenience sampling was used
and 104 patients were included based on the study’s inclusion and
exclusion criteria. Data was collected for six months i.e. from July
until December 2013. Data was then analysed using SPSS version 18.
Kaplan-Meier and Cox regression analyses were used to measure
median survival times and predictor factors for treatment failure.
The study population consisted mainly of male subjects, aged 30-
45 years, who were heterosexual, and had HIV infection for less than
6 years. The most common second line HAART regimen given was
lopinavir/ritonavir (LPV/r)-based combination. Kaplan-Meier
analysis showed that patients on LPV/r demonstrated longer median
survival times than patients on indinavir/ritonavir (IDV/r) based
combination (p
Abstract: Glutathione S-transferase was purified from human
erythrocytes and effects of some polyphenols were investigated on
the enzyme activity. The purification procedure was performed on
Glutathione-Agarose affinity chromatography after preparation of
erythrocytes hemolysate with a yield of 81%. The purified enzyme
showed a single band on the SDS-PAGE. The effects of some
poliphenolic compounds such as catechin, dopa, dopamine, progallol
and catechol were examined on the in vitro GST activity. Catechin
was determined to be inhibitor for the enzyme, but others were not
effective on the enzyme as inhibitors or activators. IC50 value -the
concentration of inhibitor which reduces enzyme activity by 50%-
was estimated to be 10 mM. Ki constants were also calculated as 6.38
± 0,70 mM with GSH substrate, and 3.86 ± 0,78 mM with CDNB
substrate using the equations of graphs for the inhibitor, and its
inhibition type was determined as non-competitive.
Abstract: Multidrug resistant organisms have been taunting the
medical world for the last few decades. Even with new antibiotics
developed, resistant strains have emerged soon after. With the
advancement of nanotechnology, we investigated colloidal silver
nanoparticles for its antimicrobial activity against Pseudomonas
aeruginosa. This organism is a multidrug resistant which contributes
to the high morbidity and mortality in immunocompromised patients.
Five multidrug resistant strains were used in this study. The
antimicrobial effect was studied using the disc diffusion and broth
dilution techniques. An inhibition zone of 11 mm was observed with
10 μg dose of the nanoparticles. The nanoparticles exhibited MIC of
50 μg/ml when added at the lag phase and the subinhibitory
concentration was measured as 100 μg/ml. The MIC50 value showed
to be 15 μg/ml. This study suggests that silver nanoparticles can be
further developed as an antimicrobial agent, hence decreasing the
burden of the multidrug resistance phenomena.
Abstract: An alarming emergence of multidrug-resistant strains
of the tuberculosis pathogen Mycobacterium tuberculosis and
continuing high worldwide incidence of tuberculosis has invigorated
the search for novel drug targets. The enzyme glutamate racemase
(MurI) in bacteria catalyzes the stereoconversion of L-glutamate to
D-glutamate which is a component of the peptidoglycan cell wall of
the bacterium. The inhibitors targeted against MurI from several
bacterial species have been patented and are advocated as promising
antibacterial agents. However there are none available against MurI
from Mycobacterium tuberculosis, due to the lack of its threedimensional
structure. This work accomplished two major objectives.
First, the tertiary structure of MtMurI was deduced computationally
through homology modeling using the templates from bacterial
homologues. It is speculated that like in other Gram-positive bacteria,
MtMurI exists as a dimer and many of the protein interactions at the
dimer interface are also conserved. Second, potent candidate
inhibitors against MtMurI were identified through docking against
already known inhibitors in other organisms.
Abstract: Today, cancer remains one of the major diseases that
lead to death. The main obstacle in chemotherapy as a main cancer
treatment is the toxicity to normal cells due to Multidrug Resistance
(MDR) after the use of anticancer drugs. Proposed solution to
overcome this problem is the use of MDR efflux inhibitor of cinchona
alkaloids which is delivered together with anticancer drugs
encapsulated in the form of polymeric nanoparticles. The particles
were prepared by the hydration method. The characterization of
nanoparticles was particle size, zeta potential, entrapment efficiency
and in vitro drug release. Combination nanoparticle size ranged 29-45
nm with a neutral surface charge. Entrapment efficiency was above
87% for the use quinine, quinidine or cinchonidine in combination
with etoposide. The release test results exhibited that the cinchona
alkaloids release released faster than that of etoposide. Collectively,
cinchona alkaloids can be packaged along with etoposide in
nanomicelles for better cancer therapy.