Captured wild specimens of the big white-toothed shrew, Crocidura lasiura were classified into three age classes by tooth wear and height of molars, and seasonal variations of breeding and reproductive organs were examined. Juveniles had not tooth wear in molars and height of the third molars were lower than the first and second molars, and had only non-breeding condition. Young adults had little tooth wear and the third molars reached to the first and second molars, and old adults had heavy tooth wear in molars, young adults and old adults had breeding or non-breeding condition according to the season. On the basis of histological examination, seasonal variations of breeding were confirmed that breeding condition of young and old adult males were continued from early February to early October although the breeding activity was the highest in April, that of females were continued from the end of March to October, males reached sexual maturity earlier than females. Whereas the breeding condition seems to cease for non-breeding season because of the deficiency of food resources, soil invertebrates. Young and old adult males of the breeding season had large testes with enlarged seminiferous tubules that were filled with numerous germ cells, and expanded caudal epididymides with a vast number of spermatozoa, and were more than 10.0 g in the body weight and 0.03 g in the testis and epididymis weight. The females of the breeding season were pregnant condition with 4~6 litters or had the Graafian follicles and the corpus lutea in the ovary, and were more than 9.6 g in the body weight.
There is growing concern that dietary soy intake is associated with protection of breast cancer. However, questions persist on the potential adverse effects of the main soy constituent genistein(GS) on female reproductive physiology. In this study, we examined whether prepubertal exposure to GS affected on the onset of puberty and the associated reproductive parameters such as hormone receptor expressions in female rats. GS(100mg/kg/day) was administrated daily from postnatal day 25(PND 25) to the day when the first vaginal opening(VO) was observed, and the animals were sacrificed on the day after VO occurred. Gross anatomy and tissue weight were compared to test the GS's effect on the cell proliferation. Furthermore, histological studies were performed to assess the structural alterations in tissues. Specific radioimmunoassay(RIA) were carried out to measure serum LH levels. To determine the transcriptional changes in progesterone receptors(PR), total RNAs were extracted from ovary and uterus and were applied to semi-quantitative reverse transcription polymerase chain reaction(RT-PCR). As a results, advanced VO was shown in the GS group(PND $31.2{\pm}0.6$) compared to the vehicle group (PND $35.3{\pm}0.7$). GS treatment significantly increased wet weight of ovaries and uteri compared to the vehicle group. Increased serum LH levels were also shown in the GS group. Graafian follicles and corpora lutea(CL) were observed only in the ovaries from GS treated animals. Similarly, hypertrophy of luminal and glandular uterine epithelium were found only in the GS group. Collectively, these effects were probably due to the estrogenic effects of GS. In the semi-quantitative RT-PCR studies, the transcriptional activities of PR in both ovary and uterus from GS-treated group were significantly higher than those from the vehicle group. The present studies demonstrated that acute exposure to GS, at levels comparable to the ranges of human exposure, during the critical period of prepubertal stage activates the reproductive system resulting precocious puberty in immature female rats.
This study was designed to examine the factors affecting in fertilization and development of embryos in vitro, and to examine whether zone drilling by laser irradiation can improve the hatching rate of IVF embryos from DNA marker-proved Hanwoo. DNA markers related to marbling score were identified using DNA fingerprinting with Ml3 probe and restriction enzyme Hae III. Oocytes were aspirated from immature ovarian follicles using a combined method of rectal ovarian-palpation and transvaginal ultrasound-guidance(6.5MHz) under local anesthesia. The aspirated oocytes were washed twice with fresh D-PBS containing 5% FBS and were rewashed 4 to 5 times with TCM-199 containing 5% FBS. A morphological grade of I to IV was assigned to each oocyte. Data were analyzed using the GLM procedure of SAS. Sperm separation methods did not have any significant effect on cleavage or developmental abilities of IVF embryos. Significantly(P<0.05) higher cleavage rate was observed in embryos from GI(60.0%, 3/5), GII(69.2%, 18/26) and GIII(62.1%, 59/95) compared to embryos from GIV oocytes(36.2%, 25/69). And the developmental rate to blastocyst stage was higher(P<0.05) in embryos from GI(33.3%, 1/3) and GII oocytes(38.9%, 7/18) than those from GIII(16.9%,10/59) and GIV oocytes(4.0%, 1/25). There was no significant difference in development of IVF embryos to blastocyst by media for in vitro culture. Proportion of hatched blastocyst was significantly(P<0.05) higher in embryos received zona drilling by laser than those of non-drilled.
Purpose: Lymphonodular hyperplasia of the colon (LNHC) is a rare finding in children and its significance as a pathologic finding is unclear. The aim of this study was to investigate the clinical significance of LNHC by analyzing clinical and histopathologic findings in children with LNHC. Methods: We analyzed data from 38 patients who were confirmed to have LNHC by colonoscopy. We checked age, birth history, past history, family history, and clinical symptoms. A hematologic exam, stool exam, and image studies were performed and biopsy specimens were examined by a pathologist. All patients were asked to have short- and long-term follow-up. Results: The mean age of the patients was 12.5${\pm}$14.4 months. All patients presented with complaints of bloody stool. They appeared healthy and the hematologic findings were within a normal range, with the exception of one case. There was no other identified source of bleeding. On histologic exam, 36 patients (94.7%) had lymphoid follicles and 34 patients (84.5%) fulfilled the criteria of allergic colitis. Regardless of diet modification and presence of residual symptom, there was no recurrence of bloody stool through long-term follow-up in all patients. Conclusion: LNHC is more common in infants who are affected by allergic colitis, but it can appear even after infancy. LNHC should be regarded as the etiology when there are any other causes of rectal bleeding, especially in healthy children. We suggest that LNHC has a benign course regardless of diet modification and it might not require excessive concerns.
The purpose of this study is to investigate the effects of the low power Helium Neon-Infra Red(HeNe-IR) laser and the special electromagnetic therapeutic apparatus stimulation, which is usually designated as TDP by using the initial of Tending Diancibo Pu which is the Chinese inscribed with English, on wound healing in rat. The seventy-five Sprague-Dawley adult female and male rats were assigned to the experimental and control groups. Each rat was anesthetized with pentobarbital sodium, and three full-thickness incisions with 12mm length wert made on the back of the half cf the rats and three deep second degree burns were made on the back of the remainder of the rats. From 34 hours after being injured, the rats of the experimental laser group were irradiated with the 157mW electric power HeNe-IR laser for 2 minutes every day and the rats of the experimental TDP group were stimulated with TDP irradiation with the 4km spot distance for 20 minutes every day during the 17 days. The rats were sacrificed and the wound parts of its were incised on the first day, 4th day, 7th day, 10th day and 17th day respectively after the beginning of wound treatment with laser and TDP irradiation. The incised wound parts were processed appropriately for the light microscopic and electron microscopic examination. The length of incised wound was measured with microcaliper before the wound part was incised. There was a significant decrease in the length of the incised wound of the experimental laser and TDP group, compared with that of control group at 4th day, 7th day and 10th day(p<0.01) after surgery. Through the histological examination of the wound site, the more rapid epithelialization and collagen formation in experimental groups were showed, compared with control group. The histological results were analyzed and summarized as the follows; The epidermis begins to be regenerated and the granulation tissue begins to be changed to the mature pattern in the H-E stained incised skin of the laser and TDP treatment group on the 4th day. The epidermis shows the complete regeneration and the granulation tissue in the dermis in mostly to be matured in the laser and TDP treatment group on the 7th day, compared with control group. The chronic inflammatory cells are oberved and the necrosis of the collagen fibers are partially observed in control group on the 10th day. The dermis of the laser and TOP treatment group reveals relatively compactly arranged collgen bundles with the mature collagen fibers on the 10th day. The epidermis and dennis of the laser and TDP group are repaired normally and the hair follicles are well regenerated on the 17th day. The mild edema and the granulation tissue is observed in the dermis of the control groups and the delayed treatment process is observed on the 17th day. The Most of proliferated collagen fibrils are found to be compact and regular in electron micrograph of burn skin of the laser treatment group on the 10th day hut the interstitial eadema and some inflammatory cells are found in the control group. The above results suggest that through the visual and histological examination the epithelized epithelium and the proliferation of the collagen liters in the dermis occur very effectively with the low power laser treatment and the TDP treatment in the incised wound healing and the burn wound healing.
Antigenic components reacting with IgE and IgG antibodies were localized in muscular layer of adult and of larva, sparganum. But the antigenic components inducing IgG were localized at tegument and parenchyma in addition to muscular layer in adult and sparganum. Also in sparganum, the surface of calcareous corpuscles of parenchyma showed immunoreactivity to IgG antibody. However antigenic components inducing IgE antibody were not localized in tegument and parenchyma, but in adult worm, we observed the immunopositive reaction at the lining of vitelline follicles in mature proglottis and on surface of egg shell within uterus of graved proglottis. By the method of immunogold-labelling, we observed the location of antigenic particles in tegument of sparganum. The density of antigenic particles inducing IgG was higher than that of antigen particles inducing IgE in syncytial tegument, tegument cells. A total of 43 and 36 protein bands were resolved from crude extracts of adult and sparganum, respectively, by SDS-PAGE. 34 bands from crude extracts of adult and larva were migrated to same positions. By EITB, 21 bands of 44 bands in adult were recognized with IgG antibody, and also 21 bands of 36 bands in sparganum. 13 bands of them were common antigenic components both in the adult worm and sparganum. Because 19 bands of 44 bands in adult worm were reacted with IgE antibody, they were IgE antigenic component. In sparganum, 13 bands were IgE antigenic components. 9 bands of them were common antigenic component inducing IgE antibody in both a-dult and sparganum. 3 bands of antigenic component recognized by IgE and IgG antibody were nonspecific antigen in both adult and sparganum of Spirometra erinacei.
Extravasation of toxic chemotherapeutic agents cause severe skin ulceration and necrosis which often need secondary surgical intervention. Still, there were not established antidote agent in case of extravasation with mitomycin-c. Dimethyl sulfoxide is known as an effective chemical scavenger of toxic hydroxyl free radical and sodium thiosulfate also was demonstrated significant protector from mitomycin-c induced ulceration by a few experimental studies. Author investigated necrotic area of mitomycin-c injected site and compare to the effectiveness of topical treatment with dimethyl sulfoxide and intradermal injection of sodium thiosulfate according to starting times, forty five mice were divided into 3 groups. Control group(n=5) had no treatment after subcutaneous injection of mitomycin-c. Experimental group I and II were 20 mice treated dimethyl sulfoxide and sodium thiosulfate, respectively. Depending on the starting time of treatment, group I and II were subdivided into 1, 2, 3 and 4 as immediate, 6 hours, 12 hours and 24 hours after mitomycin-c injection. Histologic studies of the necrotic area and survival area after treatment were performed using hematoxylin-eosin staining. The mean necrotic area of group I was significantly decreased depending on the starting time of treatment compared with control group(p<0.01). The results means there was no necrosis area which was treated with topical sodium thiosulfate within 6 hours, and it showed also significant decrease of necrosis area within 24 hours. There was also no necrosis area in group II-1 and significant decrease of necrosis area II-2 and III-3. But, effctiveness of intradermal injection of sodium thiosulfate was not found in group II-4 which was started after 24 hours. Hisotolgic findings showed a bland coagulative necrosis without inflammatory changes and no granulation tissue. The significant difference that cytoplasmic loss of subcutaneous fat and decrease number of hair follicles between two groups resulted from the methods of treatment by topical application and intradermal injection. In conclusion, immediate treatments with topical dimethyl sulfoxide or intradermal injection of sodium thiosulfate signifcantly prevents necrosis by extravasation of mitomycin-c.
This experiment was designed to evaluate the effect of transforming growth factor-$\beta$ (TGF-$\beta$) and insulin-like growth factor-I (IGF-I) in bovine oocyte maturation in the presence or absence of serum on subsequent fertilization and embryo development. In addition, various concent rations of these growth factors were evaluated for the ability to promote development of eight-cell stage embryos to the blastocyst stage. Cumulus-oocyte complexes were recovered from 2 to 6 mm follicles obtained from slaughterhouse ovaries and cultured at 38.5$^{\circ}C$ for 24 hours in TCM-199 (HEPES Modification) with or with out 20 % fetal bovine serum (FBS) to which the following growth factors were added TGF$\beta$ IGF-l or TGF $\beta$ + IGF-I, all at 10 ng/ml each. The matured oocytes were fertilized in IVF-TL medium with frozen-thawed semen at a concentration of 1 ${\times}$ 10$^6$ cells/ml of fertilization medium following Percoll separation. After 24 hours of sperm-egg incubation, the embryos were transferred to CZB medium without glucose for 48 hours and then cultured in TCM-199 with 20% fetal bovine serum (FBS) for 96 hours. The addition of growth factors to IVM medium in the presence of serum had no effect on cleavage and subsequent embryo devlopment to blastocyst. In the absence of serum, TGF- improved cleavage and development to blastocyst compared to control's(p<0.05) and no synergistic effeet of IGF-I + TGF-$\beta$ was observed. In the second experiment, eight-cell embryos obtained by in vitro maturation (IVM) in TCM-199 + 20% FBS without growth facrors and in vitro fertil-ization (IVF) were cultured in the in vitro cuiture (IVC) medium supplemented with 5, 10 ng/ml TGF-$\beta$ or 5, 10, 50, 100 ng/ml IGF-I. Cleavage rate and development to the blastocyst stage was observed during seven days of incubation. The supplementation of 10 ng/ml TGF-$\beta$ to lVC medium for eight-cell embryos improved development to blastocyst (p<0.05) compared to control. In conclusion, these data indicate that the supplementation of growth factors to IVM medium in the presence of serum does not influence cleavage and subsequent embryo development. However, significantly more oocytes matured in serum-free TCM-199 and eight-cell embryos cultured in lVC medium developed to blastocyst with supplementation of 10ng/ml TGF-$\beta$.
This study was carried out to investigate cytotoxicity of paraquat on NIH 3T3 fibroblasts, toxicity of paraquat and compensatory effects of 3-methylcholanthrene (3-MC) on the rat lung. In order to conduct MIT [3-(4,5-Dimethylthiazol-2-yl) -2,5-diphenyl -2H-tetrazolium-bromide] and NR (Neutral red) assay, the $5.0{\times}10^4cell/ml$ of NIH 3T3 fibroblast in each well of 24 multi-dish were cultured. After 24 hours, the cells were treated with solution of paraquat (1, 25, 50 and $100{\mu}M$ respectively). After the NIH 3T3 fibroblast of all groups were cultured in same condition for 48 hours. MIT and NR assay were performed to evaluate the cytotoxicity of cell organelles. $MTT_{50}\;and\;NR_{50}$ of paraquat were $1668.97{\mu}M\;and\;1030.85{\mu}M$, respectively. These $IC_{50}$ of Paraquat were decided as a low cytotoxicity by Borenfreund and Puemer (1984). In order to observe the toxicity and compensatory effects of paraquat on the rat lung, Spraque Dawley male rats were used as experimental animals and were divided into paraquat only treated group and simultaneous application group of paraquat and 3-MC, at 30 min and 1, 3, 6, 12, 24, 48 and 96 hrs interval after each treatment. The animals were sacrificed by decapitation and their or the lungs were immediately removed, immersed in fixatives, and were processed with routine method for light microscopic study. Paraffin sections were stained with H&E and iron hematoxylin of Verhoeff. Under the light microscopy, erythrocytes were full in alveolar capillaries at 3 hrs and congested at 24 hrs after paraquat administration. The great alveolar cells (Type II cell) were increased and mitosis of great alveolar were observed in interalveolar septa. Many lymphocytes, macrophages and polymorphonuclear (PMN) cells were observed in connective tissue surrounding lung tissue and germinal center in lymph follicles of terminal bronchiole. Alveolar macrophages were increased in interalveolar septa and alveoli at 48 hrs. And observed many alveolar macrophages at 96 hrs. In iron hematoxylin stain of Verhoeff, Collagen fiber were increased in respiratory bronchiole, interalveolar septa and alveoli and breath of alveoli, and alveolar pore were broaden. But, in paraquat plus 3-MC treated group, morphological changes were mild in lung tissue. These results indicate that 3-MC has a compensatory effects against toxicity of paraquat by conjugation with oxygen.
Kim, Kyu Hyon;Jung, Bum Sik;Park, Soo Bong;Park, Hang Kyun
Current Research on Agriculture and Life Sciences
/
v.8
/
pp.45-50
/
1990
This study was carried out ot investigate the effects of fetal calf serum (FCS) and gonadotropins supplemented to the medium on maturation and fertilization in vitro of porcine follcular oocytes. Ovaries were obtained from gilts at local slaughter-house. Oocyte-cumulus complexes were recovered by puncturing the ovarian follicles(3~5 mm in diameter). The complexes from individual ovaries were pooled in a $0.4m{\ell}$ droplet of medium covered with paraffin oil, then washed twice in fresh droplet and cultured for 36hrs in culture media according to experimental conditions. Boar epididymal spermatozoa were capacitated by preincubation for 4hrs in m-KRB medium and the preincubated spermatozoa were insemenated in the fertilization medium containing the cultured oocytes. The results obtained in this study are summarized as follows: 1. The maturation rates of oocytes cultured in m-KRB and m-KRB supplemented to 10% FCS were 82 and 37%, respectively. When PMSG, hCG. and PMSGt hcG($10Iu/m{\ell}$) were added to the media supplemented to 10% FCS, the maturation rates were 66, 58 and 68%, respectively. 2. Expansion of cumulus cells was not occured in m-KRB and m-KRB supplemented to 10% FCS. However, when PMSG, hCG and PMSG+hCG($10Iu/m{\ell}$) were added to m-KRB supplemented to 10% FCS, the expansion rates of cumulus cell layers were 92, 13 and 91%, respectively. 3. When oocytes were mltured in m-KRB, the rates of penetration and formation of male pronucle: were 93 and 7%, respectively. By adding FCS and gonadotropin to m-KRB, the penetration and formation of male pronuclei were 100 80%, respectively.
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