Background: Essential thrombocythemia (ET) is a clonal hemopoietic stem cell myeloproliferative neoplasm characterized by persistent thrombocytosis along with megakaryocytic hyperplasia. In the last decade following the identification of an acquired JAK2 V617F mutation, there has been acceleration in our understanding of this disease. The rational of this study was to determine the mutational profile of JAK2 V617F in Pakistan patients with ET. Materials and Methods: In this retrospective cross sectional study, 21 patients with ET were enrolled from January 2011 to December 2014. Patients were diagnosed based on WHO criteria for essential thrombocythemia. Complete blood count was done on an automated hematology analyzer, while JAK2 V617F expression was evaluated by polymerase chain reaction. Results: The mean age was $56.7{\pm}19.0$ years (range 18-87) and the male to female ratio was 1:1.1. The frequency of JAK2 V617F positivity in our ET patients was found to be 61.9%. The mean hemoglobin was $11.7{\pm}2.4$ g/dl with a total leukocyte count of $13.3{\pm}8.1{\times}109/l$ and a platelet count of $1188{\pm}522{\times}109/l$. Positive correlations for JAK2 V617F mutation were established with high TLC count and raised LDH (P<0.05). No correlation of JAK2 V617F could be established with age and gender (P>0.05). Conclusions: JAK2 V617F mutation frequency in our ET patients was similar to those reported previously. Screening for the mutation in all suspected essential thrombocythemia cases could be beneficial in differentiating patients with reactive and clonal thrombocytosis.
Purpose: The aim of this study was conducted to investigate the mean nRBC count in very low births weight infants (VLBWIs) and to determine the usefulness of the nRBC as an independent prognostic factors of perinatal complications in VLBWIs. Methods: This study was conducted on 112 VLBWIs who were hospitalized in the neonatal intensive care unit (NICU) of the author's hospital within the period from March 2003 to and May 2008. Based on the infants' nucleated red blood cells (nRBC) counts at birth, on the third day after birth, on the seventh day after birth, in the second week after birth, and in the fourth week after birth in the medical records, the correlation between nRBC or absolute nRBC counts with birth weight, gestational age, and other perinatal outcomes were retrospectively investigated. Results: In VLBWIs, their mean nRBC and absolute nRBC counts were showing a gradual decrease after birth, and they were consisteantly kept at low values since one week after and inversely proportional to the birth weights. The mean nRBC counts based on the stage after birth showed a significant correlation with perinatal death, necrotizing enterocolitis, and severe intraventricular hemorrhage. Conclusion: The increase in the nRBC count showed a significant correlation with having a severe intraventricular hemorrhage, necrotizing enterocolitis, and perinatal death in VLBWIs. If an increase or no decrease in the nRBC count after birth is observed, newborninfant care precautions should be required.
Purpose: The purpose of this study is to test the efficacy of various methods of fat harvesting in animal model by viability comparison with assay including cell counting, MTT assay, and histologic evaluation. Materials and methods: New Zealand white rabbits experiments were used. Groin fat pads were subjected to different harvest method varying ingredients of solution(Experiment 1: T1 solution= lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 10mgEq/L, Triamcinolone 10mgEq/L; T2 solution=lidocaine 1000mg/L, epinephrine 1mg/L, sodium bicarbonate 0mgEq/L, Triamcinolone 0mgEq/L) and pressure exerted on harvesting with Luer-Lock syringe connected to suction cannula.(Experiment 2: P1 group=3cc intermittent pressure; P2 group=10cc sustained pressure) Fat cell viability was assessed with cell counting with a hemocytometer, MTT assay, and histologic evaluation. Results: Experiment 1 Cell count: T1=2.4/3.4/4.2, T2=9.6/8.4/7.2($\times10^5$ per mL); MTT assay: T1=0.516/0.41/0.453/0.412/0.421, T2=0.925/0.765/0.54/0.634/0.614 in 21 days(absorbance); Histology: T1 showed elongated and, different in size and shape, and ruptured adipocytes with only a few normal adipocytes whereas T2 showed central core of fat with almost intact fat cells Experiment 2 Cell count: P1=1.2/3.2/4.2, P2=1.2/2.4/3.8($\times10^5$ per mL); MTT assay:P1=0.256/0.245/0.258/0.21/0.264, P2=0.12/0.231/0.245/0.313/0.281 in 21 days(absorbance); Histology: P1 showed somewhat evenly distributed normal-looking fat cells and P2 showed relatively irregular shape of fat cells with small blood vessel amongst adiopocytes. Conclusion: Viability was higher in ‘modified tumescent solution’without sodium bicarbonate and triamcinolone and we also found no significantly different viability between using intermittent pressure and using sustained pressure. But in terms of initial viability of fat cell, we can assume that lower intermittent pressure would make better clinical results.
Kim, Ju-Seok;Bae, Sang-Ho;Hwang, Min-Ji;Heo, Min-Yeong;Doh, Chil-Hoon
Journal of the Korean Electrochemical Society
/
v.14
no.3
/
pp.171-175
/
2011
Lithium secondary batteries using lithium metal count electrode are easy to use and to analyze the specific characteristics of working electrode. Nevertheless, during the charge operation internal electrical short circuit could be caused by the dendritic growth of lithium. The cell failure by the short circuit depends on the condition of separator such as constitutive material and thickness. To prevent the cell failure caused by the dendritic growth of lithium, the electrochemical properties of the cell of lithium metal count electrode were evaluated for four different kinds of separator. Among the tested separators, GMF (glass micro-fiber filter, $300{\mu}m$) was the most promising one because it could effectively prevent the cell failure during the charge. The cell using GMF separator had relatively low impedance. Generally the cell using thicker separator than $50{\mu}m$ could effectively avoid the cell failure by internal short circuit and had the good cycleability. The highest rate capability by the signature method was acquired in the case of GMF separator.
Park, Se-Pill;Kim, Eun-Young;Lee, Keum-Si;Lee, Young-Jae;Shin, Hyun-Ah;Min, Hyun-Jung;Lee, Hoon-Taek;Chung, Kil-Saeng;Lim, Jin-Ho
Clinical and Experimental Reproductive Medicine
/
v.29
no.2
/
pp.129-138
/
2002
Objective: This study was to compare the characteristics between parthenogenetic mES (P-mES) cells and in vitro fertilization mES cells. Materials and Methods: Mouse oocytes were recovered from superovulated 4 wks hybrid F1 (C57BL/6xCBA/N) female mice. For parthenogenetic activation, oocytes were treated with 7% ethanol for 5 min and $5{\mu}g$/ml cytochalasin-B for 4 h. For IVF, oocytes were inseminated with epididymal sperm of hybrid F1 male mice ($1{times}10^6/ml$). IVF and parthenogenetic embryos were cultured in M16 medium for 4 days. Cell number count of blastocysts in those two groups was taken by differential labelling using propidium iodide (red) and bisbenzimide (blue). To establish ES cells, b1astocysts in IVF and parthenogenetic groups were treated by immunosurgery and recovered inner cell mass (ICM) cells were cultured in LIF added ES culture medium. To identify ES cells, the surface markers alkaline phosphatase, SSEA-1, 3,4 and Oct4 staining were examined in rep1ated ICM colonies. Chromosome numbers in P-mES and mES were checked. Also, in vitro differentiation potential of P-mES and mES was examined. Results: Although the cleavage rate (${\geq}$2-cell) was not different between IVF (76.3%) and parthenogenetic group (67.0%), in vitro development rate was significantly low in parthenogenetic group (24.0%) than IVF group (68.4%) (p<0.05). Cell number count of ICM and total cell in parthenogenetic b1astocysts ($9.6{\pm}3.1,\;35.1{\pm}5.2$) were signficantly lower than those of IVF blastocysts ($19.5{\pm}4.7,\;63.2{\pm}13.0$) (p<0.05). Through the serial treatment procedure such as immunosurgery, plating of ICM and colony formation, two ICM colonies in IVF group (mES, 10.0%) and three ICM colonies (P-mES, 42.9%) in parthenogenetic group were able to culture for extended duration (25 and 20 passages, respectively). Using surface markers, alkaline phosphatase, SSEA-l and Oct4 in P-mES and mES colony were positively stained. The number of chromosome was normal in ES colony from two groups. Also, in vitro neural and cardiac cell differentiation derived from mES or P-mES cells was confirmed. Conclusion: This study suggested that P-mES cells can be successfully established and that those cell lines have similar characteristics to mES cells.
Background: Nucleolar organizer regions(NORs) are chromosomal segments encoding for ribosomal RNA and associated with argyrophilic nonhistone protein. Ribosomal RNA genes ultimately direct ribosome and protein synthesis, and it has been suggested the numbers of NORs detected in the cell may reflect nuclear and cellular activity. This study was performed to evaluate the applicability of AgNORs to the diagnosis of squamous cell carcinoma of the lung. Method: The one step silver methods(AgNORs) was used to stain NORs in the routinely processed, formalin fixed, paraffin embedded sections of 36 cases of squamous cell carcinoma of the lung obtained by surgical resection of primary tumor. In each specimen, 100 tumor cells and 100 normal cells adjacent to the tumor chosen at random were examined under an oil immersion lens at a magnification of ${\times}1000$. The mean number of AgNORs per nucleus was calculated for each specimen. Results: The mean number of AgNORs per nucleus(mAgNORs) of normal bronchial epithelium and squamous cell carcinoma of the lung was $1.74{\pm}0.25$ and $4.05{\pm}0.80$, respectively. The difference of mAgNOR between normal and tumor tissue was statistically significant(p<0.001). There was no statistical difference among tumors of different stages. The difference of mAgNOR between normal and tumor tissue was statistically significant in each TNM stage(p<0.05). Conclusion: Mean AgNOR count may be used as a useful marker for the differential diagnosis of benignancy and malignancy, and proliferative activity of the cell in squamous cell carcinoma of the lung. But there was no statistical difference in mean AgNOR count among tumors of different surgical stages. Further studies for the application of mAgNORs to the diagnosis of other histologic types and cytologic specimens of the lung cancer are needed.
Recently, there is a worldwide concern that a great number of man-made chemicals have a hormone-like action both in humans and in animals. DECD is developing screening programs using validated test systems to determine whether certain substances may have an effect in humans. In the present study. the establishment oj repeated-dose toxicity test method was tried. Flutamide. an anti-androgenic agent. was administered by gavage to Sprague-Dawley rats for 28 days at dose levels of 0. 0.5. 3 and 18 mg/kg body weight (10-15 rats/sex/group) to examine the effects on general findings. especially reproductive and endocrine parameters. Clinical signs. body weights, food consumption, and sexual cycle were checked and measured. For the gross and microscopic examinations. 10 rats/sex/group were sacrificed at the end of dosing period and the remaining animals of control and high dose groups (5 each) were sacrificed after 14 days recovery. Examinations for hematology and clinical chemistry were carried out at necropsy. There were no treatment-related changes in clinical signs. body weights, food consumption. gross necropsy. hematology and clinical chemistry at all doses of both sexes. The period and regularity of sexual cycle were not adversely affected at all doses by the test agent. At 18 mg/kg. both decreased weights of prostate, seminal vesicle and epididymis in males and increased weights of spleen and thymus in females were observed. In addition, decreased number of spermatids and sperms. increased serum testosterone concentration and increased incidence (100%) of interstitial cell hyperplasia were seen in males. At 18 mg/kg of the recovery group. decreased prostate weight. reduced sperm count and increased incidence (20%) of interstitial cell hyperplasia in males and increased thymus weight in females were observed. At 3 mg/kg. reduced sperm count was found. There were no adverse effects on parameters examined at 0.5 mg/kg of both sexes. The results suggested that the potential target organs of flutamide may be accessory sexual glands including testes for males and spleen and thymus for females. Taken together. this test method was found to be a useful screening test system for endocrine disrupting chemicals.
Yadav, N.K.;Mandal, Ajoy;Sharma, D.K.;Rout, P.K.;Roy, R.
Asian-Australasian Journal of Animal Sciences
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v.19
no.11
/
pp.1524-1528
/
2006
A total of 437 animals, comprising lambs aged between 3 and 12 months and adults of either sex of Muzaffarnagari sheep maintained at the Central Institute for Research on Goats, Makhdoom, Farah, Mathura, India were screened to assess the prevalence of Haemonchus contortus infection following natural infection and to identify the various factors affecting faecal egg count (FEC) and packed cell volume (PCV) of ewes and their genetic control. The relationships between FEC, PCV and body weight were also estimated. The prevalence rate for H. contortus infection in the flock under study was 15.7% indicating much lower occurrence of worm infection in lambs up to one year of age. On the other hand, a large proportion i.e., 67.7% of sheep was refractive to natural H. contortus infection. The random effect of sire significantly contributed (p<0.01) variation in log-transformed FEC (LFEC) of ewes. The season of birth had a significant (p<0.01) effect on LFEC of ewes. The lactating ewes had significantly (p<0.01) higher faecal egg counts compared to dry and pregnant ewes. The linear regression effects of the age of ewes on LFEC of animals were significant (p<0.01) in the present study. The heritabilities of LFEC, PCV and body weights of ewes during the course of infection were moderate to high in magnitude and ranged from 0.24 to 0.47. The LFEC of ewes was significantly (p<0.05) and negatively correlated with PCV at both genetic and phenotypic level. The genetic and phenotypic relationships between LFEC and body weights of ewes were -0.26 and -0.06 for this breed. The genetic correlation of PCV and body weight of ewes was positive and high (0.58) and statistically significant (p<0.05) but it was negatively correlated (-0.01) with body weight at the phenotypic level.
Park, Sun-Hee;Min, Byung-Soon;Choi, Ho-Young;Park, Sang-Jin;Choi, Gi-Woon
Restorative Dentistry and Endodontics
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v.16
no.2
/
pp.99-117
/
1991
The purpose of this study is to evaluate the effects of dentin bonding agents on the fibroblasts cultivated from human pulp tissue. The fibroblasts were cultured in DMEM/10%FBS medium. Whatman filter paper discs (6mm diameter) soaked with $2{\mu}l$ of dentin bonding agents were placed on a millipore filter (pore size $0.22{\mu}m$) contained in a 50mm Petri dish, and then, exposed for 10 minutes, 30 minutes, 1 hour, 6 hours, 12 hours, 24 hours, 4 days and 7 days in $37.^{\circ}C$, 5% $CO_2$ incubator. The results of the experiments were analyzed by counting the cells and measuring the protein contents at 1 day, 4 days and 7 days. The results of this study were as follows: l. CLEARFIL NEW BOND, LITE-FIL BOND, GLUMA 3 Primer and GLUMA 4 Sealer showed cytotoxicity compared to the control group in the cell counts and the protein contents. 2. GLUMA 4 Sealer showed the least cytotoxicity among the three dentin bonding agents. 3. The results of the cell count were simialr to the results of protein content measurement. 4. LITE-FIL BOND exhibited marked cytotoxicity during 1 day, but, the cytotoxicity was slightly reduced after 4 and 7 days. 5. In GLUMA 3 Primer group, it was not possible to count the cell numbers and measure the protein contents, but the degeneration of cells was observed under the inverted phase-contrast microscope.
Cytomegalovirus is the most common cause of life-threatening viral infection in HIV-infected patients. This study was done prospectively to investigate the incidence of CMV infection according to the decrease of CD4+ T cell count (CD4+) in Korean AIDS patients. Thirty-nine HIV-infected patients diagnosed before 1994 were followed for regular immunological monitoring. We have used urine shell vial method for the CMV detection from 1994 and have also checked clinical findings. Positive urine culture rate definitely depended on the CD4+ as follows; 45%, 22%, 17%, 11% and 0%, CD4+ <50, 50-100, 100-200, 200-500 and >500, respectively. Except culture positive 2 patients with CD4+ of $200{\sim}300/{\mu}l$, all eight culture positive patients with CD4+ less than $200/{\mu}l$ showed CMV related diseases on or before urine culture. But, we could not get a positive culture for a late AIDS patient with vision loss. With ganciclovir therapy, all culture results were at least negative just after or on late of first 14 days-ganciclovir infusion-course. These data suggest that the incidence of CMV disease in Korean AIDS patients is very high, and early diagnosis and treatment for CMV diseases is required for the prevention of life threatening results.
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