Serum lipid (SL) is a commercially available cholesterol-rich, proteinaceous compound extracted from bovine serum. Here we investigated the adipogenic transdifferentiation potential of SL on bovine myogenic satellite cells. Exposure of satellite cells to SL could generate lipid droplets on day 2, and further exposure to SL increased cytoplasmic lipid accumulation giving adipocyte morphology. The expression analysis of PPAR gamma and GPDH adipocyte markers along with Oil-red-O staining results confirmed the transdifferentiation potential of SL. When cells were treated at different concentrations (5, 10, 20, $40{\mu}l$/ml) of SL, the results indicated that even levels as low as $5{\mu}l$ SL /ml could induce transdifferentiation, and maximum induction was obtained at $20{\mu}l$ SL/ml. After treatment with SL at different concentrations the expression levels of PPAR gamma varied significantly (p<0.05), whereas the expression of other adipogenic transcription factors showed no difference, indicating that SL acts through PPAR gamma. The combined effect of SL and troglitazone proved to be the best combination for induction of transdifferentiation compared to the individual effect of SL or troglitazone. Thus, overall results clearly show that SL induces transdifferentiation of bovine myogenic satellite cells to adipocytes.
We investigated the antiplatelet effect of a newly synthesized guanidine derivative KR-32558, a sodium-hydrogen exchanger-1 (NHE-1) inhibitor, together with the elucidation of the possible mechanisms of action. KR-32558 concentration -dependently inhibited the aggregation of washed rabbit platelets induced by collagen (10 ${\mu}g/ml$) with an $IC_{50}$ value of 85.9 ${\mu}M$, but with much weaker potency against aggregation induced by thapsigargin (0.5 ${\mu}M$) or A23187 (5 ${\mu}M$). And had no effect on platelet aggregation induced by arachidonic acid (100 ${\mu}M$), thrombin (0.05 U/ml) and U46619 (1 ${\mu}M$) up to 100 ${\mu}M$. KR-32558 completely inhibited the $[Ca^{2+}]_i$ mobilization induced by collagen at concentration of 100${\mu}iM$. Taken together, these observation suggest that KR-32558 selectively inhibited collagen-mediated platelet aggregation by blocking the cytoplasmic calcium mobilization in addition to NHE-1 inhibition.
Ahmed, Mukhtar;Ahamed, R. Nazeer;Aladakatti, R.H.;Ghodesawar, M.G.
Advances in Traditional Medicine
/
v.9
no.4
/
pp.339-349
/
2009
An attempt has been made to assess whether the dose dependent effect of benzene extract of Ocimum sanctum leaves on the morphological changes in the cauda epididymal spermatozoa and sperm parameters in male albino rats. Scanning Electron Microscope observations illustrate the disturbance in plasma membrane as well as acrosomal membrane. Most of the sperms appear morphologically abnormal in the mid region of the tail; there is formation of balloon like cytoplasmic droplet. Sperm parametric study exhibits decrease in the total sperm count, sperm motility, forward velocity and increase in the percentage of abnormal sperms in dose dependent manner on treatment benzene extract of Ocimum sanctum leaves. The results suggest that the effects may have resulted from a general disturbance in the proteins and alteration in cauda epididymal milieu probably due to androgen deficiency consequent upon antiandrogenic property of Ocimum sanctum leaves.
To investigate an effect of the topical toluene application to .at skin on the liver injury, toluene (35 mg/$cm^2$) was sequentially applied for 3 or 5 days to rat skin and then the animals were sacrificed. 5 day toluene-treated rats showed the slight increase of live. weight per body weight(%) compared with control. Serum levels of xanthine oxidase and alanine aminotransferase activity were significantly increased both in 3 days and 5 days toluene-treated animals compared with control. In the histopathological findings, cytoplasmic degeneration of hepatocytes around the central vein was noted in the liver of rats applied with toluene to the skin. These results indicate toluene application to rat skin feds to somewhat slight liver injury. On the other hand, the hepatic benzylalcohol or aldehyde dehydrogenase activities were significantly decreased by toluene application to rat skin. In conclusion, the liver min was induced by toluene application to rat skin, and it can be hypothesized that accumulation of benzaldehyde in liver cell may be responsible for liver injury.
Alpha lipoic acid (ALA) is a biological membranes compound. As the antioxidant, it decreases the oxidized forms of other antioxidant substances such as vitamin C, vitamin E, and glutathione (GSH). To examine the effect of ALA on the in vitro maturation (IVM) of porcine oocytes, we investigated intracellular GSH and reactive oxygen species (ROS) levels, and subsequent embryonic development after parthenogenetic activation (PA). Intracellular GSH levels in oocytes treated with 50uM ALA increased significantly (P < 0.05) and exhibited a significant (P < 0.05) decrease in intracellular ROS levels compared with the control group. Oocytes matured with 50 uM of ALA during IVM displayed significantly higher cleavage rates (67.8% vs. 83.4%, respectively), and higher blastocyst formation rates and total cell number of blastocysts after PA (31.6%, 58.49 vs. 46.8%, 68.58, respectively) than the control group. In conclusion, these results suggest that treatment with ALA during IVM improves the cytoplasmic maturation of porcine oocytes by increasing the intracellular GSH levels, thereby decreasing the intracellular ROS levels and subsequent embryonic developmental potential of PA.
In the present study, mouse follicular oocytes and 2-cell embryos(late -zygote stage embryos included) were cultured on the electric pad for electric stimulation in the culture incubator. In addition, follicular oocytes and embryos were tested for maturation and development under higher temperature condition($39^{\circ}C$).Mouse follicular oocyte maturation were not affected by anion electric stimulation and there is no significant difference in GBVD and MI between the control and experiment group after 4hr culture. In the embryo culture, it was found that more morula and blastocyst were found in the electric stimulation group rather than the control(96hr). This may seem to be caused with cytoplasmic $Ca^{2+}$ transient rise by electric stimulation(anion pad). On the other hand higher temperature incubation ($39^{\circ}C$) on the anion pad caused all the embryos degenerated within $12h{\sim}24hr$ culture. This was quite different from large animal embryos(bovine, pig, sheep), in which beneficial effect of high temperature incubation for oocyte maturation and embryo development were found.
Transovarial transmission of cytoplamic polyhedrosis virus in the silkworm was studied by observing the phenomena of induction, interference and virulence enhancement in the larvae from moths inoculated with hexagonal Polyhedra of cytoplasmic polythedrosis virus. The experimental results obtained are as followings. 1. The effect of inoculation with tetragonal polyhedra of cytoplasmic polyhedrosis virus on the rate of infective induction with hexagonal polyhedron virus and with hexagonal plus tetragonal polyhedron viruses in the larvae from moths infected with hexagonal polyhedron virus was studied. Infection rate was higher by 40 to 60 percent in the larvae from infected female group than in tile larvae from noninfected female group. 2. In the studies of the effect of formalin-feeding on the induction of infection with hexagonal polyhedron virus, infection rate was higher by 40 percent in the larvae from infected female group than in the larvae from noninfected female group. However, there was no significant difference in the infection rates between the two formalin-concentration groups. 3. The effect of cold treatment on the induction of infection with hexagonal polyhedron virus was studied. Infection rate was higher by 50 percent in the larvae from infected female group than ill the larvae from noninfected female group. No difference was found in the infection rates of the two treatment groups of 12 and 48 hours. 4. The phenomena of induction and interference were studied by observing rate of infection with hexagonal polyhedron virus induced by the inoculation with tetragonal polyhedron virus. The degree of interference of primary hexagonal polyhedron virus by secondary tetragonal Ployhedron virus was increased as the dosage of secondary virus was increased. At the concentration of 1${\times}$10$\^$8/m1 of the secondary virus, the degree of interference was similar to. that of control group. On the other hand, infection with tetragonal polyhedron virus at low concentration was interfered by the primary virus. At the concentration of 1N10f/m1 of tetragonal polyhedron virus, however, the rate of infection with tetragonal polyhedron virus was increased sharply, which is still lower by 30 percent than that of control group. 5. In the studies of induction and virulence enhancement, based on the 1ate of mixed infection with hexagonal and tetragonal polyhedron viruses, the highest difference of infection rate between experimental group and control group exceeded 40 percent when the concentration of tetragonal polyhedron virus was 1${\times}$10$\^$7/m1. However, the infection rate of control group was not affected by concentrations of tetragonal polyhedron virus.
The poxvirus group is considered to be a typical cytoplasmic inclusion forming virus. Every poxvirus has been reported to produce only one kind of inclusion in the infected tissues. A vague concept that inclusions of poxviruses are eosinophilic or acidophilic has prevailed. Although many papers and theories about the nature of the inclusion have been presented, most of them are not quite convincing on the point of the relations with virus multiplication, and an analysis of papers published showed that there seem to be many discrepancies in the descriptions of the nature of the poxvirus inclusions. Comparative studies on host-virus interaction with cowpox, orf, swinepox and fowlpox viruses which selected from each Group (I-IV) of poxviruses were performed from the morphological and virological standpoints. At first, in cowpox virus-FL cell system, as a comparative model, cytoplasmic inclusion, nucleic acid metabolism by autoradiography and detection of viral antigen by immunofluorescence were studied and obtained the results as follows: 1. The focus-like cytopathic effect (CPE) at early stage developed to entire culture at terminal stage of infection, and also the developing status of CPE was correlated to viral doses for inoculation. Two kinds of cytoplasmic inclusions which named A and B type were easily observed by Giemsa, hematoxylin-eosin (H & E) and May-Greenwald Giemsa (MGG) stainings in the infected cells. The B type inclusions were formed at early stage of infection and the A type inclusions were produced subsequently the B type formation. The B type which common type inclusion in poxviruses was a small compact or aggregate at early stage and developed to a large diffuse body at terminal stage of infection. On the other hand, the A type inclusion which depend upon the kind of virus was appeared as round and discrete shape, and its size and number was increased gradually during the culture period. It was characteristic to form distinct halos around the both types of inclusions in acid fixed, H & E stained preparations of infected cultures. The B type inclusion was always positive in Feulgen reaction and showed as DNA containing body but the A type inclusion was not. 2. In the relationship between inclusion and DNA metabolism of infected cells by the qualitative autoradiography using 3H-thymidine, the appearance of silver grains was coincided with B type inclusion but not with A type inclusion. This showed that the DNA synthesis was proceeded in all B type inclusions except those in the terminal stage with a diffuse form. This suggested that the B type inclusions are only sites of DNA synthesis and this was proceeded after the cell infection independently. The activity of DNA synthesis of the inclusions was nearly the same as that of the nucleic of normal cells and non-inclusion bearing cells. and non-inclusion bearing cells. Regardless of the size of the degree of DNA synthesis of the B type inclusion, inclusion bearing cells all showed remarkable suppression of nuclear DNA synthesis. 3. By the direct fluorescent antibody technique viral antigen in infected cells was detected. The B type inclusions have been proved to contain a great deal of viral antigen, whereas the basic substance of A type inclusion did not show antigenicity except the round edge. It was suggested that the round edge fluorescence might be caused by the glare of cytoplasmic viral antigen which pushed out and concentrated by the A type inclusion development. 4. Hemorrhagic red pock formations on chorioallantoic membrane of embryonated chicken egg had proved the characteristic of used viral strain. 5. By the above studies on the nature of two types of inclusions and the role they play in virus multiplication, it was concluded that the B type inclusion must be the site of the synthesis of viral DNA and protein as well as the site of the virus.
The aim of the present study was to investigate whether ginsenoside-Rb2 (Rb2) can affect the secretion of catecholamines (CA) in the perfused model of the rat adrenal medulla. Rb2 ($3{\sim}30{\mu}M$), perfused into an adrenal vein for 90 min, inhibited ACh (5.32 mM)-evoked CA secretory response in a dose- and time-dependent fashion. Rb2 ($10{\mu}M$) also time-dependently inhibited the CA secretion evoked by DMPP ($100{\mu}M$, a selective neuronal nicotinic receptor agonist) and high $K^+$ (56 mM, a direct membrane depolarizer). Rb2 itself did not affect basal CA secretion (data not shown). Also, in the presence of Rb2 ($50{\mu}g/mL$), the secretory responses of CA evoked by veratridine (a selective $Na^+$ channel activator ($50{\mu}M$), Bay-K-8644 (an L-type dihydropyridine $Ca^{2+}$ channel activator, $10{\mu}M$), and cyclopiazonic acid (a cytoplasmic $Ca^{2+}$-ATPase inhibitor, $10{\mu}M$) were significantly reduced, respectively. Interestingly, in the simultaneous presence of Rb2 ($10{\mu}M$) and L-NAME (an inhibitor of NO synthase, $30{\mu}M$), the inhibitory responses of Rb2 on ACh-evoked CA secretory response was considerably recovered to the extent of the corresponding control secretion compared with the inhibitory effect of Rb2-treatment alone. Practically, the level of NO released from adrenal medulla after the treatment of Rb2 ($10{\mu}M$) was greatly elevated compared to the corresponding basal released level. Collectively, these results demonstrate that Rb2 inhibits the CA secretory responses evoked by nicotinic stimulation as well as by direct membrane-depolarization from the isolated perfused rat adrenal medulla. It seems that this inhibitory effect of Rb2 is mediated by inhibiting both the influx of $Ca^{2+}$ and $Na^+$ into the adrenomedullary chromaffin cells and also by suppressing the release of $Ca^{2+}$ from the cytoplasmic calcium store, at least partly through the increased NO production due to the activation of nitric oxide synthase, which is relevant to neuronal nicotinic receptor blockade.
To find out the effect of oxytocin on somatic cell count and milk production, 12 primiparous and multiparous Murrah buffaloes were selected, immediately after the parturition, from the Institute's buffalo herd. These were divided into two groups of 6 each. Buffaloes of group I did not receive oxytocin injection (control); whereas, buffaloes of group II were administered oxytocin during early lactation (av. 42.50 days). The oxytocin injection was given in doses of 2.5 IU i.m. before the start of milking, to let down the milk, for a period of 5 days. Samples of milk from individual buffaloes were collected for 5 days before (Period I), during (Period II) and after (Period III) from both the group of buffaloes. Milk samples of A. M. and P. M. milking were composited in proposition to milk yields for analysis of milk constituents. Normal values of somatic cell counts in group I of buffaloes varied from 0.54 to $0.75{\times}10^{5}cells/ml$. Mean cytoplasmic particles and epithelial cells varied from 3.68 to $7.19{\times}10^{5}cells/ml$ and 0.13 to $0.54{\times}10^{5}cells/ml$. On percentage basis the epithelial and the total leucocyte count were 60 and 40. Total leucocyte count, in the study varied from 0.17 to $0.69{\times}10^{5}cells/ml$. The differential cell count of milk indicated presence of lymphocytes (16.50 to $61.16{\times}1000$), neutrophil (0.00 to $2.00{\times}1000$) and monocyte (0.00 to $18.16{\times}1000$). Somatic cell count (p<0.01) and epithelial cells (p<0.05) varied between buffaloes and between periods of study. Total leucocyte counts of milk were also significantly varied between periods (p<0.05). The change in fat, lactose, chloride, EC and NEFA concentrations during different periods of study, were highly significant, indicated diurnal variations in different buffaloes during different days of experiment. Administration of oxytocin resulted in increase in somatic cell counts of milk (p<0.01) due to the increases in total leucocyte count (p<0.01) during the treatment period. The differential cell count indicated that oxytocin administration increased lymphocyte number significantly (p<0.01). However, secretion of neutrophil, monocyte and cytoplasmic particles were not affected by oxytocin. Eosinophil and basophil cell, though present in few samples, remain unaffected by oxytocin administration. There was no effect of oxytocin on milk production, composition, pH, EC and NEFA concentration.
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