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PubMed1988

Staining of the nucleolus with protein and RNA stains for automatic measurement of nucleolar size in paraffin sections.

In this paper two staining methods for automatic assessment of nucleolar profile area in paraffin sections of breast carcinomas using the IBAS 2000 image analyzer are reported. In the first method, the nucleolar proteins are stained with phloxine B, in the second the nucleolar RNA is stained with methyl green-pyronin. Both methods are fully reproducible. Statistically, the profile areas observed for the same patient by the two methods were found not to differ significantly. Because of their rigidity and their size in relation to section thickness, nucleoli are particularly attractive cell structures for quantitation, for instance in studies of variables that might be useful in establishing the prognosis for carcinoma patients.

Breast Neoplasms↗
PubMed2003

Fossomatic cell-counting on ewe milk: comparison with direct microscopy and study of variation factors.

Using the Fossomatic method (FSCC) a total of 23,003 analytical SCC observations were carried out on 6400 aliquots taken from 80 individual ewe milk samples with the objective of studying the influence of 4 preservation procedures (without preservation, potassium dichromate, azidiol, and bronopol), 2 storage temperatures (ambient and refrigeration), 10 milk ages (3,6,12, and 24h, and 2,3,4,5,7, and 9d postcollection), and two analytical temperatures (40 and 60 degrees C). In addition, each sample was analyzed with direct microscopic method (DMSCC), using 3 different stainings for each sample: methylene blue (MB), May-Grünwald-Giemsa (MGG) and Pyronin Y-methyl green (PGM). This allowed DMSCC and FSCC (at 24 h of age) to be compared. The reference DMSCC from MB staining was a reliable method in ewe milk, though more specific stainings such as MGG and PMG slightly improve the residual standard deviation for repeated SCC. Between DMSCC and FSCC, the highest coefficients of correlation (0.972 to 0.996) corresponded to preserved and refrigerated milk, and the lowest (0.708 to 0.919) to unpreserved and ambient stored aliquots. Except for the unpreserved and ambient stored aliquots, SCC values were similar in all aliquots. Under FSCC, preservation, storage and analytical temperature, milk age, and most of the interactions showed a significant effect on SCC variation. In preserved samples, logSCC values ranged between 5.67 (bronopol) and 5.62 (azidiol). The higest values (5.72) were for unpreserved milk, which showed false overestimation of SCC due to bacterial proliferation. LogSCC was higher at 60 degrees C (5.68) than at 40 degrees C (5.65). The interaction between age, preservation and storage temperature showed no cell degeneration in properly handled samples over the 9 d of study.

Animals↗
PubMed2004

Evaluation of rapid somatic cell counters under different analytical conditions in ovine milk.

A total of 31 individual ovine milk samples, ranging between 30 and 2600 x 10(3) cells/mL, were divided into 8 aliquots/milk with the objective of studying the overall accuracy of 2 rapid somatic cell count (SCC) counters, one based on cytometry on disk (Fossomatic 360) and the other on flow cytometry (Fossomatic 5000), under 4 types of preservation (without preservation, bronopol, sodium azide, and potassium dichromate) and 2 analytical temperatures (40 and 60 degrees C). All analyses were carried out in duplicate. In addition, each sample was analyzed in quadruplicate by reference microscopic method using Pyronin Y-methyl green as a stain. A second experiment using 13 samples divided into 20 aliquots/ sample, enabled repeatability to be studied depending on the values obtained in the SCC and on the SCC equipment used. Comparison of the methods was based on repeatability and accuracy studies (means comparison and regression studies vs. reference method). Both counters gave adequate repeatability and accuracy values in ovine milk, though the SCC obtained by Fossomatic 5000 was closer to the reference method and was somewhat more repeatable than Fossomatic 360. In the regression study, slope and intercept values were statistically different from their theoretical values (1.00 and 0.00, respectively) in the unpreserved samples but not in the preserved ones. In all cases, correlation coefficients very close to 1.00 were obtained. The preserved milk analyzed by flow cytometry gave optimal repeatability values (s(r) = 16.3 to 19.7 and s(r)% = 1.9 to 2.4), and their logSCC means (5.62 to 5.64) were not different from the reference value (5.63). Bronopol was the optimal preservative for the Fossomatic method. Analytical temperature did not contribute significantly to SCC variation, although disk cytometry gave slightly more repeatable SCC at 40 degrees than at 60 degrees C.

Animals↗
PubMed2006

Evaluation of the overall accuracy of the DeLaval cell counter for somatic cell counts in ovine milk.

The DeLaval cell counter (DCC) is a portable device designed for on-farm somatic cell count (SCC) analysis in bovine milk. This study evaluated the performance of the DCC when analyzing ovine milk. A total of 29 composite ovine milk samples, ranging between 20 x 10(3) and 2,200 x 10(3) cells/mL, were divided into 15 aliquots/milk sample corresponding to 5 SCC methods using 3 types of preservation (unpreserved, azidiol, and bronopol). The SCC methods were the Fossomatic (FSCC), the DCC in undiluted samples, and the DCC in samples diluted 1:1 in 3 different types of diluents (PBS + Triton X-100, PBS + ethidium bromide + Triton X-100, and PBS + propidium iodide + Triton X-100). All analyses were carried out in duplicate. In addition, each sample was analyzed in quadruplicate by the direct microscopic method (DMSCC) using Pyronin Y-methyl green as a stain. Comparison of methods was based on overall accuracy studies (means comparison, repeatability, and regression studies vs. DMSCC and FSCC as reference methods). The DCC methods used to analyze milk samples diluted in staining solution (with ethidium bromide or propidium iodide) showed large coefficients of regression (b = 0.91 to 1.01) and correlation (r > 0.99) when compared with the DMSCC and FSCC methods. In these samples the DCC gave repeatability values (s(r) = 33 to 48 x 10(3) cells/mL) similar to the DMSCC (s(r) = 36 x 10(3) cells/mL), and their log SCC means (5.52 to 5.54) did not differ from the reference value (5.54). However, undiluted samples analyzed by the DCC method showed large standard deviations of repeatability and SCC values lower than those by the DMSCC or FSCC methods, probably because of the high solids content in ovine milk. The type of preservation did not affect the outcomes. Consequently, the DCC was determined to be accurate when analyzing diluted ovine milk based on comparison with the SCC reference methods.

Animals↗
PubMed1993

Prevalence of high somatic cell counts in bulk tank goat milk.

Higher SCC have been observed in goat milk than in cow milk. This study evaluated the prevalence of high SCC and determined whether increased neutrophils were a factor in high SCC. Bulk tank goat milk samples were collected from commercial dairy goat herds in California, Arkansas, Michigan, and Wisconsin. Total SCC were determined using a Fossomatic Electronic Cell Counter and microscopically using pyronin Y-methyl green stain. Differential counts were determined using Wright's-stained cytospin centrifuge preparations. Average SCC were high (6.12 log10 cells/ml; geometric mean, 1.32 x 10(6) cells/ml). The average of the Fossomatic and direct microscopic SCC indicated that 8.6% of the producers had SCC < .75 x 10(6) cells/ml and that 34.5% were < 1 x 10(6) cells/ml. The current goat milk SCC standard is 1 x 10(6) cells/ml; the cow milk SCC standard will be .75 x 10(6) cells/ml in 1993. Milk samples contained 9.9% macrophages, 2.8% lymphocytes, and 87% neutrophils. These data indicate that high SCC were prevalent and that increased neutrophils contributed to high SCC in the bulk tank goat milk.

Animals↗
PubMed1976

[Studies on the lungfluke, Paragonimus iloktsuenensis: V. Host tissue reactions in albino rats]

In order to understand the tissue responses of albino rat host against Paragonimus iloktsuenensis infection, the histopathological changes of the spleen and the lungs in 6 experimental groups of rats were observed in relation with the growth, maturation and migration of this lung fluke. Rats of the experimental groups, each group consisted of 5 rats, were infected with the metacercariae of P. iloktsuenensis which were isolated from brackish water crab, Sesarma dehaani, and were kept for 3 days, 3 weeks, 4 weeks, 7 weeks, 10 weeks and 14 weeks of infection period. Peripheral blood smear slides for the differential leukocyte count were prepared and also worm collection was completed immediately after the infection period. Paraffin sections of the spleen and the lung tissue were stained with hematoxylin-eosin and methyl-green-pyronin (MGP) stain. Those materials from the experimental groups were examined in comparison with the materials obtained from control group, with special reference to immunologic aspects of host response. The results obtained were as follows: 1. The counts of large pyroninophilic cell (LPC) in the periarterial sheath of spleen were rapidly increased in earlier period of infection, and those of peribronchial lymphatic tissue started to increase after the penetration of lungfluke into the lungs. 2. The LPC counts of both the spleen and the lungs were on the decrease in conjunction with the necrosis of the lung fluke in 14th week of infection. 3. On observing differential leukocyte count of peripheral blood smear, the fluctuation of lymphocyte count was proportional to that of LPC count, and the lymphocyte count was consistently higher than that of normal rats. On the other hand, neutrophil count of experimental group showed reciprocal relation to the LPC counts. 4. The nature and characteristics of pulmonary lesion produced by the P. iloktsuenensis were just the same as those produced by P. westermani. The lesions were represented by thick and fibrosclerotic cavern, granuloma due to eggs, pneumonic process and cellular infiltrations.

Journal Article↗
PubMed1976

[Studies On The Lungfluke, Paragonimus Iloktsuenensis: VI. Effect Of Prednisolone Injection On The Immune Responses Of Albino Rats]

In order to understand the effect of prednisolone injection on the immune responses of albino rat against Paragonimus iloktsuenensis infection, the differential leucocyte counts, appearance of immunoblast (large pyroninophilic cell, LPC) in the spleen and lungs in various experimental groups were observed in relation with the growth, maturation and migration sites of this rodent lungfluke. Rats of 3 experimental groups (A series), each group consisted of 5 rats, were infected with 20 metacercariae of P. iloktsuenensis, and they were kept for 3 days(Group I), 3 weeks(Group II), and 4 weeks (Group III) of infection period. The same number of experimental groups, each group of rats received 10 mg/kg dose of prednisolone injection every other day, were also kept and examined in comparison with the former groups. Preparation of peripheral blood smear and collection of worms were completed immediately after the end of infection period, and they were stained with Giemsa or Semichon's acetocarmine. Paraffin sections of the spleen and the lung tissues were stained with hematoxylin-eosin and methyl green pyronin (MGP). Those materials from A and B series of experimental groups were examined under the light microscope, and the results obtained were as follows: 1. On observing differential leucocyte counts of peripheral blood smear, lymphocyte counts were consistently higher than those of uninfected controls in A series of infected groups, while those of B series were consistently low. On the other hand, neutrophil counts of A series showed lower counts than those of B series. In general, fluctuation patterns of both A and B series of experimental groups were almost the same, although lymphocyte and neutrophil counts showed reciprocal relation. The eosinophil counts of both series were negligible, especially in the groups of B series. 2. The counts of LPC in the periarterial lymphatic sheath of the spleen were rapidly increased in the groups of A series, while those of B series were much less than those of A series, and the appearance of considerable LPC in the spleen was also delayed in B series. Furthermore, LPC of peribronchial lymphatic tissue in A series started to increase after the invasion of lungflukes into the lungs, while those of B series were much less due to the inhibited migration of lymphocytes into the lesions. 3. Number, size and maturity of collected worms showed no significant differences between the groups of A and B series, but migration speed of the lungflukes was somewhat accelerated in B series than in A series. In this connection, it was considered that the immune responses of albino rats did not contribute for the complete protection against P. iloktsuenensis, but inhibited the migration of this lungfluke to some extent.

Journal Article↗
PubMed1977

[Histochemical studies on Echinorhynchus gadi (Acanthocephala)]

The distribution of glycogen, polysaccharide, mucopolysaccharide, lipid and nucleic acid has been studied in Echinorhynchus gadi(Acanthocephala). The results were summarized as follows: 1. Glycogen and polysaccharide was demonstrated by Bauer PAS reaction technique and was found in fertilization membrane in ovum, central nuclear mass in acanthor and lemnisci, hypodermis in cystacanth. 2. Mucopolysaccharide was demonstrated by Mowry alcian blue staining technique and was found in outer membrane, fibrillar coat, fertilization membrane and inner membrane in acanthocephalan ova. 3. Lipid was demonstrated by Smith Nile blue stain and Lison Sudan black B staining technique and was found roughly parallel to that of polysaccharide. 4. Nucleic acid was demonstrated by Rosenbeck Feulgen reaction, Taft methylgreen-pyronin stain and Diengdoh acridine orange staining technique and found in central nuclear mass in acanthor, also, was found in lemnisci, proboscis and hypodermis in cystacanth.

Journal Article↗
PubMed1992

[Squamous metaplasia of the mucosa of the human laryngeal ventricle: a clinicopathological study with reference to the possibility of de novo carcinogenesis].

Primary carcinomas are rarely encountered in the laryngeal ventricle of Morgagni. They are classified into the subgroup of supraglottic laryngeal carcinoma in the TNM classification system of malignant tumors. However, detailed investigations have not hitherto been made on the possible occurrence of carcinomas in this region of the larynx. To find out whether the laryngeal ventricle could be the site of carcinogenesis, the author examined squamous metaplasia in the mucosal epithelium of 60 human laryngeal ventricles which were taken from 38 larynges removed operatively from patients with laryngeal carcinoma (14 cases), hypopharyngeal carcinoma (20 cases), carcinoma of the upper esophagus (two cases), tracheal carcinoma (one case) or thyroid carcinoma (one case). The patients consisted of 31 males and 7 females, with an age distribution of 44 to 80, average 62.7. The laryngeal ventricles were selected for investigation only when they did not show any carcinomatous invasion macroscopically. Macroscopic observations were performed after gross staining with pyronin Y, which permits differentiation of squamous metaplasia from respiratory epithelia. Tissues were embedded in paraffin, sectioned serially and stained with hematoxylin and eosin. The clinical records of the patients were also made. The results are summarized as follows: 1. Squamous metaplasia of the stratified ciliated epithelium was found in 38 out of 60 specimens (63%) of the laryngeal ventricles examined. 2. The metaplasia seemed to be precipitated by smoking or exposure to radiation, along with aging. 3. The incidence and extent of metaplasia of the ventricular mucosa depended on the disease entity which had necessitated total laryngectomy, and the metaplasia was most prominent in cases with the supraglottic type of laryngeal carcinoma. 4. The metaplastic epithelium was occasionally associated with hyperplasia or dysplasia of the epithelium. 5. Microinvasive carcinoma was found in one laryngeal ventricle with severe metaplasia. These findings suggested a possible de novo carcinogenic process in the laryngeal ventricle through squamous metaplasia of the respiratory epithelium. From this observation, the author proposes to call carcinoma of this region of the larynx the ventricular type of laryngeal carcinoma.

Adult↗
PubMed1999

Isolation of a highly quiescent subpopulation of primitive leukemic cells in chronic myeloid leukemia.

Chronic myeloid leukemia (CML) is characterized by an increased proliferative activity of the leukemic progenitors that produce an elevated number of mature granulocytes. Nevertheless, cell cycle-active agents, even in very high doses, are alone unable to eradicate the leukemic clone, suggesting the presence of a rare subset of quiescent leukemic stem cells. To isolate such cells, we first used Hoechst 33342 and Pyronin Y staining to obtain viable G(0) and G(1)/S/G(2)/M fractions of CD34(+) cells by fluorescence-activated cell sorting (FACS) from 6 chronic-phase CML patients' samples and confirmed the quiescent and cycling status of the 2 fractions by demonstration of expected patterns of Ki-67 and D cyclin expression. Leukemic (Ph(+)/BCR-ABL(+)) cells with in vitro progenitor activity and capable of engrafting immunodeficient mice were identified in the directly isolated G(0) cells. Single-cell reverse transcriptase-polymerase chain reaction (RT-PCR) analysis showed that many leukemic CD34(+) G(0) cells also expressed BCR-ABL mRNA. CD34(+) from 8 CML patients were also labeled with carboxyfluorescein diacetate succinimidyl diester (CFSE) before being cultured (with and without added growth factors) to allow viable cells that had remained quiescent (ie, CFSE(+)) after 4 days to be retrieved by FACS. Leukemic progenitors were again detected in all quiescent populations isolated by this second strategy, including those exposed to a combination of flt3-ligand, Steel factor, interleukin-3, interleukin-6, and granulocyte colony-stimulating factor. These findings provide the first direct and definitive evidence of a deeply but reversibly quiescent subpopulation of leukemic cells in patients with CML with both in vitro and in vivo stem cell properties.

Animals↗
PubMed1975

Studies with Brugia pahangi. III: Histological changes in the affected lymph nodes of infected cats.

Cats infected with Brugia pahangi by single or repeated inoculation of infective larvae for different periods of time, were autopsied and the affected lymph nodes examined histologically, stained with methyl green and pyronin Y, haematoxylin and eosin, Lendrum's eosinophil stain, Mallory's trichrome stain or the May-Grunwald Giemsa technique. In single infections, there was initially a cell-mediated type immunological response which was characterized by proliferation of mononuclear thymus-dependent cells in the paracortical region of the lymph node. The antibody-type response became more evident as the infections progressed. This response was characterized by proliferation of lymphocytes in rapidly enlarging germinal centres and the production of large numbers of plasma cells in the cortico-medullary junction and medullary cords of the nodes. In long term infections, the affected afferent lymphatic vessel and popliteal node were almost obliterated by fibrous tissue, but an inguinal lymph node complex usually developed to replace lymphatic filtration in the limb. Repeated infections produced more complicated changes. Both types of response occurred to varying degrees throughout the infection. No significant increase in mast cells or eosinophil cells was seen. Fibrous tissue increased in the lymph nodes only after long periods of infection; such nodes often became non-functional, and by-passed by new lymphatic vessels.

Animals↗
PubMed2000

[Estimation of selected methods of separation of hematopoietic cells from human umbilical cord blood].

Umbilical cord blood can determine alternative to bone marrow source of hematopoietic cells for transplantations. The aim of this work was estimation and comparison both of effectivity of separation of mononuclear cells from umbilical cord blood by means of Ficoll-Histopaque, Gradisol L and ammonium chloride and potential toxicity of Hoechst 33342, Rhodamin 123 and Pyronin Y on CD34+ cells. We showed superiority of density gradient centrifugation method (Ficoll-Histopaque, Gradisol L) over hypoosmotic lysis of erythrocytes and granulocytes (ammonium chloride). We ascertained similar efficiency of Ficoll and Gradisol. We showed also significant toxic influence of metabolic dyes, especially Hoechst 33342 on early hematopoietic cells.

Cell Separation↗
PubMed2000

Human hematopoietic stem cells stimulated to proliferate in vitro lose engraftment potential during their S/G(2)/M transit and do not reenter G(0).

An understanding of mechanisms regulating hematopoietic stem cell engraftment is of pivotal importance to the clinical use of cultured and genetically modified transplants. Human cord blood (CB) cells with lymphomyeloid repopulating activity in NOD/SCID mice were recently shown to undergo multiple self-renewal divisions within 6 days in serum-free cultures containing Flt3-ligand, Steel factor, interleukin 3 (IL-3), IL-6, and granulocyte colony-stimulating factor. The present study shows that, on the fifth day, the transplantable stem cell activity is restricted to the G(1) fraction, even though both colony-forming cells (CFCs) and long-term culture-initiating cells (LTC-ICs) in the same cultures are approximately equally distributed between G(0)/G(1) and S/G(2)/M. Interestingly, the G(0) cells defined by their low levels of Hoechst 33342 and Pyronin Y staining, and reduced Ki67 and cyclin D expression (representing 21% of the cultured CB population) include some mature erythroid CFCs but very few primitive CFCs, LTC-ICs, or repopulating cells. Although these findings suggest a cell cycle-associated change in in vivo stem cell homing, the cultured G(0)/G(1) and S/G(2)/M CD34(+) CB cells exhibited no differences in levels of expression of VLA-4, VLA-5, or CXCR-4. Moreover, further incubation of these cells for 1 day in the presence of a concentration of transforming growth factor beta(1) that increased the G(0)/G(1) fraction did not enhance detection of repopulating cells. The demonstration of a cell cycle-associated mechanism that selectively silences the transplantability of proliferating human hematopoietic stem cells poses both challenges and opportunities for the future improvement of ex vivo-manipulated grafts. (Blood. 2000;96:4185-4193)

Animals↗
PubMed2000

[Immunohistochemical study of interstitial keratitis in an animal model].

PURPOSE: We performed an immunohistochemical study on the development of interstitial keratitis in a rabbit model sensitized by ovalbumin (OA). METHODS: A mixture of OA (5 micrograms/ml) and Freund's complete adjuvant was initially injected subcutaneously (0.4 ml) into the rabbits' back and footpad (0.2 ml). After a week, the rabbits were sensitized by an injection of the same solution into the back (0.5 ml) again. After 4 weeks the rabbits underwent the same treatment as a booster shot. Then a week later OA solution (0.05 ml) was injected into the corneal stroma. We observed the cornea on days 1-3, 7, and 10 after the corneal treatment. Corneas on days 3 and 10 were examined by immunohistological methods using hematoxylin-eosin stain, methylgreen pyronin stain, and by immunohistochemical methods with anti-CD 4, anti-CD 8, and anti-OA antibodies. The localization of specific antibodies was identified by fluorescent-antigen method. Electron-microscopic observation was also done. RESULTS: Rabbits developed corneal opacity and edema on day 1, corneas had on immune ring with a peak intensity on day 3, and vascularization on day 7 after corneal treatment. Microscopic examination revealed infiltration of neutrophils in to the area of the immune ring on day 3 and many plasma cells at the limbus and stroma on day 10. CD 4 positive cells were found at the limbus and stroma on days 3 and 10. CD 8 positive cells were found at the limbus and stroma only on day 10. OA positive findings were observed at the immune ring and its inner area. In the fluorescent-antigen method, specific antibodies were found in plasma cells at the limbus on days 3 and 10. CONCLUSIONS: In our experimental model of interstitial keratitis with immune ring and vascularization, the Arthus reaction was dominant. Infiltration of antigen specific plasma cells in the stroma with vascularization caused a modification of pathologic reaction in keratitis.

Animals↗
PubMed1975

[Cytochemical study of the elimination chromatin in silkworm meiosis].

The elimination chromatin separating from chromosomes during the first maturation division of female sex cells of lepidoptera insects was studied cytochemically on paraffin sections of eggs of Bombyx mori L. Ocytes at the metaphase-telophase stage of the first meiotic division were stained for RNA by metyl green-pyronin and gallocyanin with negative results. These data differ from earlier positive results for Solenobia triquetrella reported by Ris and Kleinfeld, 1952.

Animals↗
PubMed2000

[The effect of dactinomycin on retinal injury induced by argon laser in mouse].

OBJECTIVE: To determine that the administration of RNA synthesis inhibitor (dactinomycin) prevents retinal photoreceptor cell injury induced by green wavelength of argon laser in SD mice. METHODS: At various time (0, 8, 20 h) after the exposure to laser, eyes of the mice were injected intraocularly with dactinomycin in experimental eyes, and saline in control ones, and the concentration of the drug were 10 g/L, 5 g/L, 2.5 g/L. The eyes were taken at 24 h after the exposure, the slices of the retina were stained with methyl green-pyronin (MG-P), and pyknotic (violet) as well as surviving cells (green) were counted in the laser burns. RESULTS: Intraocular administration of dactinomycin diminished the degeneration induced by argon laser, there were marked differences between the experimental group and the control group, dactinomycin showed to be more effective if administered immediately after exposure and 8 h later, after this time the higher concentration of the drug could not result in higher effect. CONCLUSION: Dactinomycin can inhibit the occurrence of the laser injury, but the time selected for the administration is critical for its efficacy.

Animals↗
PubMed2002

Antiapoptotic effect of (-)-deprenyl in rat kidney after ischemia-reperfusion.

BACKGROUND: Since apoptosis of renal tubular cells is the basis of the damage caused by ischaemia-reperfusion, the antiapoptotic effect on kidney tubular epithelial cells of the monoamino oxidase-B (MAO-B) inhibitor (-) deprenyl (selegiline), known neuroprotective agent, with antiapoptotic properties, was studied in a rat model. MATERIAL/METHODS: Warm renal ischaemia was caused by clamping the left renal artery of rats for 30 minutes. With the start of reperfusion 0.015 mg/kg, 0.15 mg/kg and 1.5 mg/kg of (-)-deprenyl was injected simultaneously into the carotid artery of the animals, respectively. Five rats served as control, in which renal artery clamping was performed, but the rats were only treated with the solvent (physiological saline). After 6 hours of reperfusion the rats were exsanguinated and the kidneys were histologically examined. RESULTS: Severe tubular damage characterised by apoptosis was found in the kidneys of the untreated rats. Apoptosis was verified on the basis of morphological features, methylgreen-pyronin staining and TUNEL reaction. (-)-Deprenyl diminished dose-dependently the apoptotic damage, 0.15 mg/kg being the most effective dose. The same dose of (-)-Deprenyl is used in the therapy of human Parkinson's disease. CONCLUSIONS: Our findings suggest, that (-)-deprenyl might have an impact on decreasing renal injury also in case of human cadaveric renal transplantation.

Animals↗
PubMed1999

[Effects of homocysteine on cardiovascular development in early chicken embryo].

OBJECTIVE: To study the relationship between homocysteine (HCY) and cardiovascular development in the early embryo and to determine whether folic acid and cobalamine (Vit B(12)) can reduce the developmental toxicity of HCY to cardiovascular system. METHODS: Incubation test for chicken embryo, electron microscopy, methylgreen-pyronine stain, in situ fragment end labeling, and intervention trial with folic acid and vitamin B(12) were used in the study. RESULTS: Treatment of 0 - 16.0 micromol of HCY/per embryo could disturb their heart development and differentiation of blood vessels at fetal ages of two and four days, in a dose-response pattern, with 24.1% and 25.0% of heart defect and 60.7% of inhibition of blood vessels of yolk sac in 8.0 micromol of HCY, respectively. A dose of more than four micromol/per embryo could damage the structure of myocardial cells and organelle, and inhibit the synthesis of cellular DNA and RNA. It was the first time to find that 8.0 micromol of HCY could induce excessive apoptosis of myocardial cells (2.7%), significantly higher than that in the normal control group. Folic acid could antagonize the cardiovascular toxicity caused by HCY, stronger than what Vit B(12) could do. CONCLUSION: HCY was a new risk factor which could damage cardiovascular development, and folic acid could effectively antagonize its developmental toxicity. Cellular apoptosis might relate to cardiovascular defect.

Animals↗