Abstract: More than 3000 plants of notable phyto-therapeutic
value grow in South Africa; these include Cissampelos capensis,
commonly known in Afrikaans as dawidjie or dawidjiewortel. C.
capensis is the most significant and popular medicinal plant used by
the Khoisan as well as other rural groups in the Western region of
South Africa. Its rhizomes are traditionally used to treat male fertility
problems. Yet, no studies have investigated the effects of this plant or
its extracts on human spermatozoa. Therefore, this study aimed at
investigating the effects of C. capensis rhizome extract (CRE)
fractions on ejaculated human spermatozoa in vitro. Spermatozoa
from a total of 77 semen samples were washed with human tubular
fluid medium supplemented with bovine serum albumin (HTF-BSA)
and incubated for 2 hours with 20 μg/ml progesterone (P4) followed
by incubation with different concentrations (0, 0.05, 0.5, 5, 50, 200
μg/ml) of fractionated CRE (F1=0% MeOH, F2=30% MeOH,
F3=60% MeOH and F4=100% MeOH) for 1.5 hours at 37°C. A
sample without addition of CRE fractions served as control. Samples
were analyzed for sperm motility, reactive oxygen species (ROS),
DNA-fragmentation, acrosome reaction and capacitation. Results
showed that F1 resulted in significantly higher values for ROS,
capacitation and hyper-activation compared to F2, F3, and F4 with
P4-stimulated samples generally having higher values. No significant
effect was found for the other parameters. In conclusion, alkaloids
present in F1 of CRE appear to have triggered sperm intrinsic ROS
production leading to sperm capacitation and acrosome reaction
induced by P4.
Abstract: Morphological interaction of porcine cumulus-oocyte
complexes (pCOCs) was investigated on in vitro condition using
electron microscope (SEM and TEM). The totals of 1,923 oocytes
were round in shape, surrounded by Zona pellucida with layer of
cumulus cells ranging between 59.29-202.14 μm in size. They were
classified into intact-, multi-, partial cumulus cell layer oocyte, and
completely denuded oocyte, at the percentage composition of 22.80%
32.70%, 18.60%, and 25.90 % respectively. The pCOCs classified as
intact- and multi cumulus cell layer oocytes were further culturing at
37°C with 5% CO2, 95% air atmosphere and high humidity for 44 h
in M199 with Earle’s salts supplemented with 10% HTFCS, 2.2
mg/mL NaHCO3, 1 M Hepes, 0.25 mM pyruvate, 15 μg/mL porcine
follicle-stimulating hormone, 1 μg/mL LH, 1μg/mL estradiol with
ethanol, and 50 μg/mL gentamycin sulfate. On electron microscope
study, cumulus cells were found to stick their processes to secrete
substance from the sac-shape end into Zona pellucida of the oocyte
and also communicated with the neighboring cells through their
microvilli on the beginning of incubation period. It is believed that
the cumulus cells communicate with the oocyte by inserting the
microvilli through this gap and embedded in the oocyte cytoplasm
before secreting substance, through the sac-shape end of the
microvilli, to inhibit primary oocyte development at the prophase I.
Morphological changes of the complexes were observed after
culturing for 24-44 h. One hundred percentages of the cumulus layers
were expanded and cumulus cells were peeling off from the oocyte
surface. In addition, the round-shape cumulus cells transformed
themselves into either an elongate shape or a columnar shape, and no
communication between cumulus neighboring cells. After 44 h of
incubation time, diameter of oocytes surrounded by cumulus cells
was larger than 0 h incubation. The effect of hormones in culture
medium is exerted by their receptors present in porcine oocyte. It is
likely that all morphological changes of the complexes after hormone
treatment were to allow maturation of the oocyte. This study
demonstrated that the association of hormones in M199 could
promote porcine follicle activation in 44 h in vitro condition. This
culture system should be useful for studying the regulation of early
follicular growth and development, especially because these follicles represent a large source of oocytes that could be used in vitro for cell
technology.
Abstract: Interaction of Schiff base complexes of Iron and
Manganese: Iron [N, N’ Bis (5- (triphenyl phosphonium methyl)
salicylidene) -1, 2 ethanediamine) chloride, [Fe Salen]Cl; Manganese
[N, N’ Bis (5- (triphenyl phosphonium methyl) salicylidene) -1, 2
ethanediamine) acetate, were investigated by spectroscopic and
isothermal titration calorimetry techniques (ITC).
The absorbance spectra of complexes have shown hyper and
hypochromism in the presence of DNA that is indication of
interaction of complexes with DNA. The linear dichroism (LD)
measurements confirmed the bending of DNA in the presence of
complexes.
Furthermore, Isothermal titration calorimetry experiments
approved that complexes bound to DNA on the base of both
electrostatic and hydrophobic interactions. More, ITC profile exhibits
the existence of two binding phases for the complexes. Antibacterial
activity of ligand and complexes were tested in vitro to evaluate their
activity against the gram positive and negative bacteria.
Abstract: Enterococci are important inhabitants of the animal
intestine and are widely used in probiotic products. A probiotic strain
is expected to possess several desirable properties in order to exert
beneficial effects. Therefore, the objective of this study was to
isolate, characterize and identify Enterococcus sp. from chicken cecal
and fecal samples to determine potential probiotic properties.
Enterococci were isolated from chicken ceca and feces of thirty three
clinically healthy chickens from a local farm. In vitro studies were
performed to assess antibacterial activity of the isolated LAB (using
agar well diffusion and cell free supernatant broth technique against
Salmonella enterica serotype Enteritidis), survival in acidic
conditions, resistance to bile salts, and their survival during simulated
gastric juice conditions at pH 2.5. Isolates were identified by
biochemical carbohydrate fermentation patterns using an API 50
CHL kit and API ZYM kits and by sequenced 16S rDNA. An isolate
belonging to E. faecium species exhibited inhibitory effect against S.
enteritidis. This isolate producing a clear zone as large as 10.30 mm
or greater and was able to grow in the coculture medium and at the
same time, inhibited the growth S. enteritidis. In addition, E. faecium
exhibited significant resistance under highly acidic conditions at pH
2.5 for 8 h and survived well in bile salt at 0.2% for 24 h and showing
ability to survive in the presence of simulated gastric juice at pH 2.5.
Based on these results, E. faecium isolate fulfills some of the criteria
to be considered as a probiotic strain and therefore, could be used as a
feed additive with good potential for controlling S. Enteritidis in
chickens. However, in vivo studies are needed to determine the safety
of the strain.
Abstract: The rhizome of Java grass, Cyperus rotundus was
extracted different organic polar and non-polar solvents and
performed the in vitro antiviral and immunostimulant activities
against White Spot Syndrome Virus (WSSV) and Vibrio harveyi
respectively. Based on the initial screening the ethyl acetate extract of
C. rotundus was strong activities and further it was purified through
silica column chromatography and the fractions were screened again
for antiviral and immunostimulant activity. Among the different
fractions screened against the WSSV and V. harveyi, the fractions, FIII
to FV had strong activities. In order to study the in vivo influence
of C. rotundus, the fractions (F-III to FV) were pooled and delivered
to the F. indicus through artificial feed for 30 days. After the feeding
trail the experimental and control diet fed F. indicus were challenged
with virulent WSSV and studied the survival, molecular diagnosis,
biochemical, haematological, and immunological parameters.
Surprisingly, the pooled fractions (F-IV to FVI) incorporated diets
helped to significantly (P
Abstract: Doxorubicin, also known as Adriamycin, is an
anthracycline class of drug used in cancer chemotherapy. It is used in
the treatment of non-Hodgkin’s lymphoma, multiple myeloma, acute
leukemia, breast cancer, lung cancer, endometrium cancer and ovary
cancers. It functions via intercalating DNA and ultimately killing
cancer cells. The major side effects of doxorubicin are hair loss,
myelosuppression, nausea & vomiting, oesophagitis, diarrhea, heart
damage and liver dysfunction. The minor modifications in the
structure of compound exhibit large variation in the biological
activity, has prompted us to carry out the synthesis of sulfonamide
derivatives. Sulfonamide is an important feature with broad spectrum
of biological activity such as antiviral, antifungal, diuretics, antiinflammatory,
antibacterial and anticancer activities. Structure of the
synthesized compound N-(1-methyl-2-oxo-2-N-methyl anilinoethyl)
benzene sulfonamide confirmed by proton nuclear magnetic
resonance (1H NMR),13C NMR, Mass and FTIR spectroscopic tools
to assure the position of all protons and hence stereochemistry of the
molecule. Further we have reported the binding potential of
synthesized sulfonamide analogues in comparison to doxorubicin
drug using Auto Dock 4.2 software. Computational binding energy
(B.E.) and inhibitory constant (Ki) has been evaluated for the
synthesized compound in comparison of doxorubicin against Poly
(dA-dT).Poly (dA-dT) and Poly (dG-dC).Poly (dG-dC) sequences.
The in vitro cytotoxic study against human breast cancer cell lines
confirms the better anticancer activity of the synthesized compound
over currently in use anticancer drug doxorubicin. The IC50 value of
the synthesized compound is 7.12 μM whereas for doxorubicin is 7.2
μM.
Abstract: Although, arsenic trioxide has been the subject of
toxicological research, in vitro cytotoxicity and genotoxicity studies
using relevant cell models and uniform methodology are not well
elucidated. Hence, the aim of the present study was to evaluate the
cytotoxicity and genotoxicity induced by arsenic trioxide in human
keratinocytes (HaCaT) using the MTT [3-(4, 5-dimethylthiazol-2-yl)-
2,5-diphenyltetrazolium bromide] and alkaline single cell gel
electrophoresis (Comet) assays, respectively. Human keratinocytes
were treated with different doses of arsenic trioxide for 4 h prior to
cytogenetic assessment. Data obtained from the MTT assay indicated
that arsenic trioxide significantly reduced the viability of HaCaT cells
in a dose-dependent manner, showing an IC50 value of 34.18 ± 0.6
μM. Data generated from the comet assay also indicated a significant
dose-dependent increase in DNA damage in HaCaT cells associated
with arsenic trioxide exposure. We observed a significant increase in
comet tail length and tail moment, showing an evidence of arsenic
trioxide -induced genotoxic damage in HaCaT cells. This study
confirms that the comet assay is a sensitive and effective method to
detect DNA damage caused by arsenic.
Abstract: Ocimum americanum L (Lamiaceae) is an annual herb
that is native to tropical Africa. The in vitro and in vivo antioxidant
activity of its aqueous extract was carefully investigated by assessing
the DPPH radical scavenging activity, ABTS radical scavenging
activity and hydrogen peroxide radical scavenging activity. The
reducing power, total phenol, total flavonoids and flavonols content
of the extract were also evaluated. The data obtained revealed that the
extract is rich in polyphenolic compounds and scavenged the radicals
in a concentration dependent manner. This was done in comparison
with the standard antioxidants such as BHT and Vitamin C. Also, the
induction of oxidative damage with paracetamol (2000 mg/kg)
resulted in the elevation of lipid peroxides and significant (P < 0.05)
decrease in activities of superoxide dismutase, glutathione
peroxidase, glutathione reductase and catalase in the liver and kidney
of rats. However, the pretreatment of rats with aqueous extract of O.
americanum leaves (200 and 400 mg/kg) and silymarin (100 mg/kg)
caused a significant (P < 0.05) reduction in the values of lipid
peroxides and restored the levels of antioxidant parameters in these
organs. These findings suggest that the leaves of O. americanum have
potent antioxidant properties which may be responsible for its
acclaimed folkloric uses.
Abstract: Hot aqueous and methanol extracts of the two selected
herbal medicines such are Vellarugu chooranam (V.C) and
Amukkirai Chooranam (A.C) were examined for total phenolic and
flavonoid contents and in vitro antioxidant activity using four
different methods. The total phenolic and flavonoid contents in
methanol extract of V.C were found to be higher (44.41±1.26mg
GAE/g; 174.44±9.32mg QE/g) than in the methanol extract of A.C
(20.56±0.67mg GAE/g; 7.21±0.85mg QE/g). Hot methanol and
aqueous extracts of both medicines showed low antioxidant activity
in DPPH, ABTS, and FRAP methods and Iron chelating activity not
found at highest possible concentration. V.C contains higher
concentrations of total phenolic and flavonoid contents than A.C and
can also exert greater antioxidant activity than A.C, although the
activities demonstrated were lower than the positive control Trolox.
The in vitro antioxidant activity was not related with the total
phenolic and flavonoid contents of the methanol and aqueous extracts
of both herbal medicines (A.C and V.C).
Abstract: Immunomodulators are substances that alter immune
system via dynamic regulation of messenger molecules. It can be
divided into immunostimulant and immunosuppressant. It can help to
increase immunity of people with a low immune system, and also can
help to normalize an overactive immune system. Aim of this study is
to investigate the effects of in vitro exposure to low and high doses of
several immunomodulators which include caffeine, kaloba and
quercetin on antigen-stimulated whole blood culture cytokine
production. Whole blood samples were taken from 5 healthy males
(age: 32 ± 12 years; weight: 75.7 ± 6.1 kg; BMI: 24.3 ± 1.5 kg/m2)
following an overnight fast with no vigorous activity during the
preceding 24 h. The whole blood was then stimulated with 50 μl of
100 x diluted Pediacel vaccine and low or high dose of
immunomodulators in the culture plate. After 20 h incubation (5%
CO2, 37°C), it was analysed using the Evidence Investigator to
determine the production of cytokines including IL-2, IL-4, IL-10,
IFN-γ, and IL-1α. Caffeine and quercetin showed a tendency towards
decrease cytokine production as the doses were increased. On the
other hand, an upward trend was evident with kaloba, where a high
dose of kaloba seemed to increase the cytokine production. In
conclusion, we found that caffeine and quercetin have potential as
immunosuppressant and kaloba as immunostimulant.
Abstract: Multipotent mesenchymal stromal cells (MSCs)
possess immunomodulatory properties. The effect of MSCs on the
crucial cellular immunity compartment – T-cells is of a special
interest. It is known that MSC tissue niche and expected milieu of
their interaction with T- cells are characterized by low oxygen
concentration, whereas the in vitro experiments usually are carried
out at a much higher ambient oxygen (20%). We firstly evaluated
immunomodulatory effects of MSCs on T-cells at tissue-related
oxygen (5%) after interaction implied cell-to-cell contacts and
paracrine factors only. It turned out that MSCs under reduced oxygen
can effectively suppress the activation and proliferation of PHAstimulated
T-cells and can provoke decrease in the production of
proinflammatory and increase in anti-inflammatory cytokines. In
hypoxia some effects were amplified (inhibition of proliferation, antiinflammatory
cytokine profile shift). This impact was more evident
after direct cell-to-cell interaction; lack of intercellular contacts could
revoke the potentiating effect of hypoxia.
Abstract: This work studied the ability of adipose tissue-derived
mesenchymal stromal cells (MSCs) to form stroma for expansion of
cord blood hematopoietic cells. We showed that 72-hour interaction
of MSCs with cord blood mononuclear cells (MNCs) in vitro at
atmospheric (20%) and low (5%) O2 conditions increased the
expression of ICAM-1, HCAM (at the beginning of interaction) on
MSCs. Viability of MSCs and MNCs were maintained at high level.
Adhesion of MNCs to MSCs was faster at 20% O2. MSCs promoted
the proliferation of adhered MNCs to form the suspension containing
great number of hematopoietic colony-forming units, and this effect
was more pronounced at 5% O2. Thus, adipose-derived MSCs
supplied sufficient stromal support to cord blood MNCs both at 20%
and 5% О2, providing their adhesion with further expansion of new
generation of different hematopoietic lineages.
Abstract: Amyloid aggregation of polypeptides is related to a
growing number of pathologic states known as amyloid disorders. In
recent years, blocking or reversing amyloid aggregation via the use of
small compounds are considered as two useful approaches in
hampering the development of these diseases. In this research, we
have compared the ability of several manganese-salen derivatives, as
synthetic compounds, and apigenin, as a natural flavonoid, to inhibit
of hen egg-white lysozyme (HEWL) aggregation, as an in vitro
model system.
Different spectroscopic analyses such as Thioflavin T (ThT) and
Anilinonaphthalene-8-sulfonic acid (ANS) fluorescence, Congo red
(CR) absorbance along with transmission electron microscopy were
used in this work to monitor the HEWL aggregation kinetic and
inhibition. Our results demonstrated that both type of compounds
were capable to prevent the formation of lysozyme amyloid
aggregation in vitro. In addition, our data indicated that synthetic
compounds had higher activity to inhibit of the β-sheet structures
relative to natural compound. Regarding the higher antioxidant
activities of the salen derivatives, it can be concluded that in addition
to aromatic rings of each of the compounds, the potent antioxidant
properties of salen derivatives contributes to lower lysozyme fibril
accumulation.
Abstract: In the present study, presence of endosulfan, imidacloprid, carbendazim, in the soil /vegetables/cereals and water samples was observed in agriculture fields of Uttarakhand. In view of biodegradation of these pesticides, 9 bacterial isolates were recovered from the soil samples of the fields which tolerated endosulfan, imidacloprid, carbendazim from 100 to 200 µg/ml. Three bacterial consortia used for in vitro bioremediation experiments were consisted of 3 bacterial isolates for carbendazim, imidacloprid and endosulfan, respectively. Maximum degradation (87 and 83%) of α and β endosulfan respectively was observed in soil slurry by consortium. Degradation of Imidacloprid and carbendazim under similar conditions was 88.4 and 77.5% respectively. FT-IR analysis of biodegraded samples of pesticides in liquid media showed stretching of various bonds. GC-MS of biodegraded endosulfan sample in soil slurry showed the presence of nontoxic intermediates. A pot trial with Bacterial treatments lowered down the uptake of pesticides in onion plants.
Abstract: Solid lipid nanoparticles (SLNs) have gained great attention for the topical treatment of skin associated fungal infection as they facilitate the skin penetration of loaded drugs. Our work deals with the preparation of nystatin loaded solid lipid nanoparticles (NystSLNs) using the hot homogenization and ultrasonication method. The prepared NystSLNs were characterized in terms of entrapment efficiency, particle size, zeta potential, transmission electron microscopy, differential scanning calorimetry, rheological behavior and in vitro drug release. A stability study for 6 months was performed. A microbiological study was conducted in male rats infected with Candida albicans, by counting the colonies and examining the histopathological changes induced on the skin of infected rats. The results showed that SLNs dispersions are spherical in shape with particle size ranging from 83.26±11.33 to 955.04±1.09 nm. The entrapment efficiencies are ranging from 19.73±1.21 to 72.46±0.66% with zeta potential ranging from -18.9 to -38.8 mV and shear-thinning rheological Behavior. The stability studies done for 6 months showed that nystatin (Nyst) is a good candidate for topical SLN formulations. A least number of colony forming unit/ ml (cfu/ml) was recorded for the selected NystSLN compared to the drug solution and the commercial Nystatin® cream present in the market. It can be fulfilled from this work that SLNs provide a good skin targeting effect and may represent promising carrier for topical delivery of Nyst offering the sustained release and maintaining the localized effect, resulting in an effective treatment of cutaneous fungal infection.
Abstract: Two new metal-based anticancer chemotherapeutic
agents, [(Ph2Sn)2(HGuO)2(phen)Cl2] 1 and [(Ph3Sn)(HGuO)(phen)]-
Cl.CH3OH.H2O 2, were designed, prepared and characterized by
analytical and spectral (IR, ESI-Mass, 1H, 13C and 119Sn NMR)
techniques. The proposed geometry of Sn(IV) in 1 and 2 is distorted
octahedral and distorted trigonal-bipyramidal, respectively. Both 1
and 2 exhibit potential cytotoxicity in vitro against MCF-7, HepG-2
and DU-145 cell lines. The intrinsic binding constant (Kb) values of 1
(2.33 × 105 M-1) and 2 (2.46 × 105 M-1) evaluated from UV-Visible
absorption studies suggest non-classical electrostatic mode of
interaction via phosphate backbone of DNA double helix. The Stern-
Volmer quenching constant (Ksv) of 1 (9.74 × 105 M-1) and 2 (2.9 ×
106 M-1) determined by fluorescence studies suggests the groove
binding and intercalation mode for 1 and 2, respectively. Effective
cleavage of pBR322 DNA is induced by 1.Their interaction with
DNA of cancer cells may account for potency.
Abstract: This study was aimed to investigate the effect of
various organic supplements on growth and development of
Dendrobium discolor’s protocorms and seedlings growth of
Dendrobium Judy Rutz. Protocorms of Dendrobium discolor with 2.0
cm. in diameter and seedlings of Dendrobium Judy Rutz at the same
size (0.5 cm. height) were sub-cultured on Hyponex medium
supplemented with cow milk (CM), soy milk (SM), potato extract
(PE) and peptone (P) for 2 months. The protocorms were developed
to seedlings in all treatments after cultured for 2 months. However,
the best results were found on Hyponex medium supplemented with
P was the best in which the maximum fresh and dry weight and
maximum shoot height were obtained in this treatment statistically
different (p ≤ 0.05) to other treatments. Moreover, Hyponex medium
supplemented with P also stimulated the maximum mean number of
5.7 shoots per explant which also showed statistically different (p ≤
0.05) when compared to other treatments. The results of growth of
Dendrobium Judy Rutz seedlings indicated the medium
supplemented with 100 mL/L PE enhanced the maximum fresh and
dry weigh per explants with significantly different (p ≤ 0.05) in fresh
weight from other treatments including the control medium without
any organic supplementation. However, the dry weight was not
significantly different (p ≤ 0.05) from medium supplemented with
SM and P. There was multiple shoots induction in all media with or
without organic supplementation ranging from 2.6 to 3 shoots per
explants. The maximum shoot height was also obtained in the
seedlings cultured on medium supplemented with PE while the
longest root length was found in medium supplemented with SM.
Abstract: This study aimed to investigate effect of different organic supplements on growth of Vanda and Mokara seedlings. Vanda and Mokara seedlings approximately 0.2 and 0.3 cm. in height were sub-cultured onto VW supplemented with 150 ml/L coconut water, 100 g/L potato extract, 100 g/L ‘Gros Michel’ banana (AAA group) and 100 g/L ‘Namwa’ banana (ABB group). The explants were sub-cultured onto the same medium every month for 3 months. The best medium increased stem height to 0.52 and 0.44 Cm. in Vanda and Mokara respectively was supplemented with coconut water. The maximum fresh weight of Vanda (0.59 g) was found on medium supplemented with ‘Gros Michel’ banana while Mokara cultured on medium supplemented with Potato extract had the maximum fresh weight (0.27 g) and number of roots (5.20 roots/shoot) statistically different (p≤ 0.05) to other treatments. However, Vanda cultured on medium supplemented with ‘Namwa’ banana had the maximum number of roots (3.80 roots/shoot). Our results suggested that growth of different orchid genera was responded diversely to different organic supplements.
Abstract: In this work we describe the preparation of NanoSilver/methylcellulose hydrogel containing silver nanoparticles (NPs) for topical bactericidal applications. Highly concentrated dispersion of silver NPs as high as of 5g/L of silver with diameter of 10nm was prepared by reduction of AgNO3 via strong reducing agent NaBH4. Silver NPs were stabilized by addition of sodium polyacrylate in order to prevent their aggregation at such high concentration. This way synthesized silver NPs were subsequently incorporated into methylcellulose suspension at elevated temperature resulting in formation of NanoSilver/methylcellulose hydrogel when temperature cooled down to laboratory conditions. In vitro antibacterial activity assay proved high bactericidal and fungicidal efficiency of silver NPs alone in the form of dispersion as well as in the form of hydrogel against broad spectrum of bacteria and yeasts including highly multiresistant strains such as methicillin-resistant Staphylococcus aureus. A very low concentrations of silver as low as 0.84mg/L Ag in as-prepared dispersion gave antibacterial performance. NanoSilver/methylcellulose hydrogel showed antibacterial action at the lowest used silver concentration equal to 25mg/L. Such prepared NanoSilver/methylcellulose hydrogel represent promising topical antimicrobial formulation for treatment of burns and wounds.
Abstract: Muscid flies are known to be vectors of disease agents and species that annoy humans and domesticated animals. An example of these flies is Musca domestica (house fly) whose adult and immature stages occur in a variety of filthy organic substances including household garbage and animal manures. They contribute to microbial contamination of foods. It is therefore imperative to control these flies as a result of their role in Public health. The second and third instars of Musca domestica (Linn) were infected with varying cell loads of Bacillus subtilis in vitro for a period of 48 hours to evaluate its larvicidal activities. Mortality of the larvae increased with incubation period after treatment with the varying cell loads. Investigation revealed that the second instars larvae were more susceptible to treatment than the third instars treatments. Values obtained from the third instar group were significantly different (P