Jang, Gook Chan;Yang, Eun Seok;Moon, Kyung Rye;Park, Young Bong;Rho, Young Il
Clinical and Experimental Pediatrics
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v.50
no.8
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pp.761-766
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2007
Purpose : Post-lumbar puncture headache is common complaint. A study of post-diagnostic lumbar puncture headache in children is rare. Various factors that might influence the occurrence of postdiagnostic lumbar puncture headache in children exist. The purpose of this prospective study was to assess the frequency and risk factors for post-diagnostic lumbar puncture headache in children. Methods : From March 2005 to February 2006, 44 patients with suspected meningitis were enrolled. Patients were received diagnostic lumbar puncture at the Chosun University Hospital, Gwangju, Korea. We evaluated age, sex, previous headache history, number of puncture attempts, volume of cerebrospinal fluid (CSF), pressure of CSF, cell count in CSF, final diagnosis, and the frequency and duration of headaches. Results : Of the 44 patients (mean age $7.36{\pm}2.04$, range 4-13 years), 16 patients (36.4%, male 13/33, 39.4%, female 3/11, 27.2%) had headache. The frequency of headaches was significantly higher in patients with previous headache history compare to those without previous headache history (P= 0.037). The mean of cell count of CSF was significantly higher in patients with post-lumbar puncture headache (P=0.012). The other factors did not influence the post-diagnostic lumbar puncture headache. Conclusion : Post-diagnostic lumbar puncture headache in children was more common than other studies. The factors that influence post-diagnostic lumbar puncture headache in children are previous headache history and cell count in CSF.
This study was designed to investigate the effects of superovulation on the growing and mature follicles following gonadotrophin treatments in mature rat by immunohistochemical methods. Eighteen mature rats (Sprague-Duwely, 190~230gm) were randomly alloted into 3 groups. One group was control group, another FSH-treated group was injected intramuscularly with 0.5 units of follicular stimulating hormone(FSH) / rat, and third PMS and HCG-treated group was injected intramuscularly with 20~25IU of pregnant mare serum(PMS) / rat and then at the 48 hrs later, with 20~25IU of human chorionic gonadotropin(HCG) / rat. Half the number of rats were administrated intraperitoneally with bromodeoxyuridine(Brdur, 0.2mg/gm BW once) at 2 hours before exanguination and the remainder of rats were sacrified without Brdur administration. The investigation by immunohistochemical methods using paraffin sections of ovaries was performed by using anti-Brdur antibody and PCNA(proliferating cell nuclear antigen) antibody for labeling proliferating cells in follicles. In immunohistochemical findings, follicles squeezed by peripheral corpus luteum or follicles large follicles with loosly and irregularly distributed granulosa cells and although with compacted granulosa cells, middle follicles with dilated round or oval follicular antrum were confirmed as atretic follicles. The proportions of atretic follicles in control group were 29.8%, 21.7% and 14.2% respectivley at large, middle and small follicles and mean proportions of these all 3 grade follicles were 26.7%. The proportions of atretic follicles in FSH-treated group were 35.4%, 24.9% and 10.4% respectively at large, middle and small follicles and mean proportions of these all 3 grade follicles were 28.1%. The proportions of atretic follicles in PMS and HCG-treated group were 44.7%, 24.0% and 12.7% respectively at large, middle and small follicles, and mean proportions of these all 3 grade follicles were 29.7%. The above findings reveal that the group with higher proportion of atretic follicles were ordered as large, middle and small follicles in size, and these proportions were increased in hormone treated two groups with more number of more growing and mature follicles when compared with control group.
Objective: The present study investigates pre- and post-implantation developmental competence of nuclear-transferred porcine embryos derived from male and female fetal fibroblasts. Methods: Male and female fetal fibroblasts were transferred to in vitro-matured enucleated oocytes and in vitro and in vivo developmental competence of reconstructed embryos was investigated. And, a total of 6,789 female fibroblast nuclear-transferred embryos were surgically transferred into 41 surrogate gilts and 4,746 male fibroblast nuclear-transferred embryos were surgically transferred into 25 surrogate gilts. Results: The competence to develop into blastocysts was not significantly different between the sexes. The mean cell number of female and male cloned blastocysts obtained by in vivo culture ($143.8{\pm}10.5$ to $159.2{\pm}14.8$) was higher than that of in vitro culture of somatic cell nuclear transfer (SCNT) groups ($31.4{\pm}8.3$ to $33.4{\pm}11.1$). After embryo transfer, 5 pregnant gilts from each treatment delivered 15 female and 22 male piglets. The average birth weight of the cloned piglets, gestation length, and the postnatal survival rates were not significantly different (p<0.05) between sexes. Conclusion: The present study found that the sex difference of the nuclear donor does not affect the developmental rate of porcine SCNT embryos. Furthermore, postnatal survivability of the cloned piglets was not affected by the sex of the donor cell.
The obseuations on the seasonal fluctuations of phytoplankton community in Haechang Bay of the Korean southern sea were carried out during four seasons from 1997 to 1998. A total of 77 species of phytoplankton including 10 freshwater species, belonging to 51 Benera was identified. Seasonal succession of dominant species was evident in Haechang Bay; Chaetoceros cunisetus, Skeletonema costatum, Eurampia zodiacus, Dictyocha fibula and Ceratium furca in summer, C. curisetus in autunm, C. cunisetus Rhizosoienia setigera and E. zodiacus in winter and S. costatum in spring. The phytoplankton community in Haechang Bay showed various species composition and was occupied with centric diatoms all the year round, Densities of the phytoplankton cell number by the samples of Haechang Bay ranged from $8.4{\times}10^3\;cells/l\;to\;2.0{\times}10^5\;cells/l$ with the mean value of $9.2{\times}10^4\;cells/l$ in summer, from $3.2{\times}10^3\;cells/l\;to\;4.6{\times}10^6\;cells/l$ with mean of $6,2{\times}10^4\;cells/l$ in autumn, from $8.4{\times}10^3\;cells/l\;to\;4.3{\times}10^4\;cells/l$ with mean $2.2{\times}10^4\;cells/l$ in winter and from $1.0{\times}\;10^3\;cells/l to\;4.6{\times}10^4\;cells/l$, with mean of $1.1{\times}10^4\;cells/l$ in spring. Phytoplankton standing crops fluctuated with an annual mean of $4.7{\times}10^4 cells/l$ between the lowest value of $1.0{\times}10^3\;cells/l$ in spring and the highest value of $4.6{\times}10^5 cells/l$ in autumn, That is, phytoplankton standing crops was high in summer and autumn, while it was very low in winter and spring.
Kim, Bo-Hyun;Kim, Ji-Su;Ryu, Jae-Sung;Lee, So-Hyun;Lee, Ju-Taek;Kang, Jae-Yul;Chang, Kyu-Tae;Choo, Young-Kug
Animal cells and systems
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v.15
no.4
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pp.287-294
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2011
The accelerated cooling rate associated with vitrification reduces injuries attributed to cryopreservation and improves the post-freezing developmental competence of vitrified embryos. In this study, embryos were vitrified and warmed and morphologically evaluated for their development to blastocysts. Survival rates between the fresh ($96.7%{\pm}3.8%$) and vitrified embryos ($90.7%{\pm}5.1%$) did not differ significantly (P>0.05). The mitochondrial membrane potential of fresh control cells measured by 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolyl carbocyanide iodide staining was similar to that of cryoprotected and vitrified embryos. Mitochondrial staining with rhodamine 123 did not differ among the fresh, cryoprotected, and vitrified embryos. Moreover, the distribution of $H_2O_2$, assessed by 2',7'-dichlorodihydrofluorescein diacetate staining, did not differ among the groups. The results showed that the developmental rate did not differ significantly among the fresh ($87.8%{\pm}11.3%$), cryoprotected ($83.2%{\pm}7.6%$), and vitrified 2-cell embryos ($75.8%{\pm}14.2%$). The mean number of the inner cell mass (ICM), trophectoderm (TE), and apoptotic cells was counted and statistically compared, and although the number of ICM and TE was decreased in the cryoprotected and vitrified embryos, there were no significant differences among the groups (P>0.05). During the cultivation period, randomly selected blastocysts from each group were stained using either 4',6-diamidino-2-phenylindole and bisbenzimide or the terminal deoxynucleotidyl transferase mediated dUTP nick-end labeling technique. The incidence of apoptosis appeared to be almost identical in all the groups. Droplet vitrification could subsequently lead to high survival and developmental rates of cryopreserved mouse embryos.
Mesna has been used with ifosfamide to prevent urotoxicity in the treatment of testicular cancers. This drug also protected the toxicities of adriamycin without compromising cytostatic activity. With an idea of radioprotective role of sulfhydryl group of radioprotectors and of mesna decreasing the toxic effect of adriamycin which produces free radicals, mesna and radiation were administered to mice to study the protective effect of this drug and to identify the difference in regenerative capacity of the germ cells in the testis between radiation-treated and both mesna-and radiation-treated groups. The shape and numbers of spermatogenic cells in the seminiferous tubules were examined every week after irradiation. In both groups, initial reduction and later recovery in germ cell numbers and shape was observed. The lowest germ cell number was found around three weeks after irradiation. Mean germ cell number of the mesna-treated group was significantly higher than radiation-treated group at all observed periods (p<0.05). More competent regeneration was present in mesna-treated group. These results suggest that mesna protect the testis from radiation injury. Further study will be necessary to identify whether mesna protects other tissues from radiation and it does not hamper tumor control.
Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.
The objective of this study was to determine the effect of fructose that was supplemented to a chemically defined in vitro maturation (IVM) medium on oocyte maturation and embryonic development after parthenogenesis in pigs. The base medium for in vitro maturation (IVM) was porcine zygote medium (PZM) that was supplemented with 0.05% (w/v) polyvinyl alcohol (PVA) or 10% (v/v) porcine follicular fluid (pFF). In the first experiment, when immature pig oocytes were matured in a chemically defined medium that was supplemented with 5.5 mM glucose or with 1.5, 3.0 and 5.5 mM fructose, 3.0 mM fructose resulted in a higher nuclear maturation (91.5%) than 1.5 and 5.5 mM fructose (81.9 and 81.9%, respectively) but showed a similar result with 5.5 mM glucose (94.2%). However, there was no significant differences among groups in the embryo cleavage (89.4-92.4%), blastocyst formation (37.5-41.1%), and mean cell number of blastocyst (30.8-34.2 cells). Fructose at the concentration of 3.0 mM (1.08 pixels/oocyte) resulted in a higher intra-oocyte glutathione (GSH) content than 1.5 and 5.5 mM fructose (1.00 and 0.87 pixels/oocytes, respectively) while the cumulus cell expansion was not influenced. In the second experiment, effect of individual and combined supplementation of a chemically defined maturation medium with 5.5 mM glucose or 3.0 mM fructose was examined. No significant effect was found in the nuclear maturation (86.3-92.6%). Embryo cleavage was significantly increased by the combined supplementation with glucose and fructose (95.2%) compared to that with 3.0 mM fructose only (85.7%) while blastocyst formation (37.3-42.8%) and embryonic cell number (33.3-34.1 cells) were not altered. Effect of supplementation of pFF-containing medium with glucose and fructose + glucose was examined in the third experiment. No significant effect by the supplementation with glucose and fructose or glucose alone was observed in the nuclear maturation of oocytes (90.7-94.1%) and blastocyst formation (51.0-56.5%). Our results demonstrate that 3.0 mM fructose was comparable to 5.5 mM glucose in supporting in vitro oocyte maturation and embryonic development after parthenogenesis and could be used as an alternative energy source to glucose for in vitro maturation of pig oocytes.
The beneficial effect of glucose and phosphate ions in culture medium on the development of human embryos in vitro has not been fully elucidated. The purpose of this study was to evaluate the influence of fertilization and culture of embryos in glucose/phosphate-free m-TALP medium on pregnancy rates in IVF-ET program. The patients in 244 IVF-ET cycles received GnRH agonist + HMG regimens. A does of 10,000 IU HCG was administered when two or more dominent follicles reached 18mm in diameter. Thirty-six hours after HCG, oocytes were recovered transvaginally using ultrasound guidance. Aspirated oocytes were matured for 4 to 6 h in TCM-199 supplemented with 10% follicular fluid (FF). Insemination was carried out with 50,000 motile spermatozoa in TCM-199 + 10% FF or m-TALP + 5% FF + 5% fetal cord serum (FCS) according to experimental design. After 6 h, oocytes were washed 3 to 4 times and cultured in each fresh medium. After 20 h, oocytes were freed from cumulus/corona cells and examined for the presence of pronuclei. Fertilized oocytes were transferred into each co-culture drops and cultured for further incubation. On day 3, embryo transfer was performed with grade 1 and 2 embryos. Monolayers for co-culture of embryos were prepared by plating $1{\times}10^5$ cumulus cells/ml in 10ul drop of TCM-199 + 10% FF or m-TALP + 5% FF + 5% FCS media 24 h prior to the onset of co-culture. Development to 4 to 16 cell stage was observed at 70x magnification following two days of incubation. Pregnancy was confirmed by detecting increasing serum ${\beta}$-hCG concentrations for 11 days following embryo transfer. Data were analyzed by ${\chi}^2$-test. Oocytes from 244 IVF-ET cycles were randomized. The number of cycles and mean age of patients were 97 and 147, 31.3 yrs and 31.2 yrs for TCM-199 (control) and m-TALP groups, respectively. The mean number of retrieved oocytes/cycle, fertilization rates, number of embryos transferred/ET and pregnancy rates were 11.1 and 10.3, 65.1% and 67.3%, 4.1 and 4.7, 28.9% and 43.8% for TCM-199 and m-TALP groups, respectively. Differences in the pregnancy rates were found between control and m-TALP groups (p<0.05). The pregnancy rate of patients divided according to maternal age groups of ${\leq}30$, 31-35, $36{\leq}$ were 44.4% and 49.0%, 26.1% and 41.3%, 29.2% and 41.2% for control and m-TALP groups, respectively. These data indicate that culture of human embryos in glucose/phosphate-free m-TALP medium improves pregnancy rates.
The clarify the ecological properties, and to predict change of understory vegetation of mt. Nam Park, population dynamics and interspecific competition of D. smilacinum and D. viridescens, which grow in understory of deciduous broad-leaved forest and pseudo-annuals, were studied from May 20 to May 30 1998. The depth of litter layer, soil moisture content, soil organic matter and soil texture were surveyed in 18 populations (15 D. smilacinum populations and 3 D. viridescens populations). Mean litter layer of d. smilacimum population was thinner than that of D. viridescens populations). Mean litter layer of D. smilacnum population was thinner than that of D. viridescens population. The contents of soil moisture and organic matter of D. smilacinum population were lower than that of D. viridescens population. The D. smilacinum growed in broad range of soil texture but D. viridescens in loamy soil. Because D. smilacinum could tolerate more broad range of soil moisture and soil texture than D. viridescens, the former covered the herb layer in earlier stage and the latter introduced in later stage when rhizome could grow easily. The numbers of individual in two marginal parts were smaller than that in center in same D. smilacinum patch. And the total numbers of individuals grown in (10 ${\times}$ 10)cm were from 0 to 12. The rhizome (subterranean runner) weight, rhizome length, root weight, shoot weight, lea weight and leaf number per subquadrat (cell) increased along the number of individual, that is, increased from marginal part to center. But rhizome weight and rhizome length per individual were vice versa. Therefore, the individuals in marginal part reproduced longer and stronger asexual propagules than that in center. The distribution pattern of D. smilacinum was contageous and that of D. viridescens was random or regular. Therefore, population growth of former was independent on density and that of latter was dependent on density. The distributions of size-class showed normal curves in two population, but the curves based on data of total dry weight showed positive skewness and those of leaf number showed negative skewness The correlation coefficient (CC) values between the properties of each organ were high in two population and significant at 0.1% level. The CC values of D. viridescens were higher of the two. Therefore, the former allocated the energy to each organ stable. The rhizome depth of d. viridescens was 2 times deeper than that of D. smilacinum. And rhizome length and weight of D. viridescens were longer (2 times) or heavier (4 times) than those of D. smilacinum. The patch size of D. viridescens increased 60 cm per year and that of D. smilacinum 30 cm. On this results, the intrinsic increase velocity of d. viridescens patch was 2 times faster than that of d. smilacinum, therefore, on the competition, the former had an advantage over D. smilacinum. The reason why d. viridescens defeated D. smilacinum resulted from that the leaf area of former was 4 times broader than that of latter. in Mt. Nam Park, it was thought that two disporum Population would change with the 3 thpes of environmental change as followings. First, no human impact and increase of soil moisture content resulted in increase of D. viridescens population. Second, mild human impact and similar condition of soil moisture content resulted in slow increase or no changes of D. smilacinum and d. viridescens population. Third, severe human impact and dry condition resulted in decrease or vanishment of two disporum populations.
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