Activation of OCT4 enhances ex vivo expansion of human cord blood hematopoietic stem and progenitor cells by regulating HOXB4 expression.

Although hematopoietic stem cells (HSC) are the best characterized and the most clinically used adult stem cells, efforts are still needed to understand how to best ex vivo expand these cells. Here we present our unexpected finding that OCT4 is involved in the enhancement of cytokine-induced expansion capabilities of human cord blood (CB) HSC. Activation of OCT4 by Oct4-activating compound 1 (OAC1) in CB CD34(+) cells enhanced ex vivo expansion of HSC, as determined by a rigorously defined set of markers for human HSC, and in vivo short-term and long-term repopulating ability in NSG mice. Limiting dilution analysis revealed that OAC1 treatment resulted in 3.5-fold increase in the number of SCID repopulating cells (SRCs) compared with that in day 0 uncultured CD34(+) cells and 6.3-fold increase compared with that in cells treated with control vehicle. Hematopoietic progenitor cells, as assessed by in vitro colony formation, were also enhanced. Furthermore, we showed that OAC1 treatment led to OCT4-mediated upregulation of HOXB4. Consistently, siRNA-mediated knockdown of HOXB4 expression suppressed effects of OAC1 on ex vivo expansion of HSC. Our study has identified the OCT4-HOXB4 axis in ex vivo expansion of human CB HSC.

Placenta-derived mesenchymal stem cells improve memory dysfunction in an Aß1-42-infused mouse model of Alzheimer's disease.

Mesenchymal stem cells (MSCs) promote functional recoveries in pathological experimental models of central nervous system (CNS) and are currently being tested in clinical trials for neurological disorders, but preventive mechanisms of placenta-derived MSCs (PD-MSCs) for Alzheimer's disease are poorly understood. Herein, we investigated the inhibitory effect of PD-MSCs on neuronal cell death and memory impairment in Aß1-42-infused mice. After intracerebroventrical (ICV) infusion of Aß1-42 for 14 days, the cognitive function was assessed by the Morris water maze test and passive avoidance test. Our results showed that the transplantation of PD-MSCs into Aß1-42-infused mice significantly improved cognitive impairment, and behavioral changes attenuated the expression of APP, BACE1, and Aß, as well as the activity of ß-secretase and γ-secretase. In addition, the activation of glia cells and the expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were inhibited by the transplantation of PD-MSCs. Furthermore, we also found that PD-MSCs downregulated the release of inflammatory cytokines as well as prevented neuronal cell death and promoted neuronal cell differentiation from neuronal progenitor cells in Aß1-42-infused mice. These data indicate that PD-MSC mediates neuroprotection by regulating neuronal death, neurogenesis, glia cell activation in hippocampus, and altering cytokine expression, suggesting a close link between the therapeutic effects of MSCs and the damaged CNS in Alzheimer's disease.

Anti-thymocyte globulin for conditioning in matched unrelated donor hematopoietic cell transplantation provides comparable outcomes to matched related donor recipients.

Rabbit anti-thymocyte globulin (ATG) is used as prophylaxis against GVHD following allogeneic hematopoietic cell transplantation (HCT). At our institution, ATG is exclusively used in the conditioning of matched unrelated donor (URD) transplant recipients. A total of 50 URD HCT recipients who received ATG (ATG group) were retrospectively compared with 48 matched related donor (MRD) HCT recipients who did not receive ATG (no ATG group). There were no significant differences between the groups in rates of day 100 mortality, acute GVHD or relapse. Chronic GVHD incidence was significantly lower in the ATG group (P = 0.007). At a median follow-up of 36 months in the entire cohort, 50% patients are alive in the ATG group and 63% of the patients are alive in the no ATG group (P = 0.13). We conclude that the administration of ATG to patients undergoing URD HCT preserves the anti-leukemia benefit of the transplant, while reducing the risk of developing GVHD, resulting in OS rates that are comparable to MRD HCT recipients.

Palliative care in BMT.

The field of high-dose therapy and SCT has made many advances in the past several years and the success rates have been steadily increasing as newer therapies emerge and improvements in supportive care continue to improve patient survival and cure rates. There still remains a mortality risk for high-dose therapy and the need for palliative care becomes more apparent as the focus also incorporates quality of life in all facets of cancer treatment and care. This paper reports on the lack of literature available on palliative care into the BMT and explores areas of future research in the integration of these two fields of medicine.

Yam bean seed poisoning mimicking cyanide intoxication.

BACKGROUND: Yam bean is a common food in southern Taiwan. However, its seeds are rarely consumed. We describe five patients of yam bean seed poisoning in Taiwan, one of them life-threatening. CLINICAL PRESENTATION: The five patients presented with perioral numbness, nausea and vomiting after eating a same soup made from yam bean seeds. One of them, a 54-year-old woman, had difficulty breathing and lost consciousness. Physical examination showed dilated pupils and coma with no focal neurological signs. The initial blood pressure was normal. Laboratory data showed a severe anion gap metabolic acidosis, with a serum lactate level of 185 mg/dL. An initial diagnosis of cyanide intoxication was considered and she was given sodium nitrite and sodium thiosulfate i.v. Hypotension ensued shortly afterwards and pulmonary artery catheterization showed a decreased cardiac index. Aggressive fluid and inotropic therapy were given and the patient eventually recovered. The other four patients suffered only minor gastrointestinal and neurological symptoms and received supportive treatment. Cyanide levels were negative in all five patients. CONCLUSION: Yam bean seed poisoning can cause acute metabolic acidosis and altered mental status, which could be confused with acute cyanide intoxication from a cyanogenic glycoside-containing plant. To our knowledge, this is the first outbreak of yam bean seed poisoning reported in the English published work.

A preliminary report of brain edema in patients with uremia at first hemodialysis: evaluation by diffusion-weighted MR imaging.

BACKGROUND AND PURPOSE: The dynamics of brain-water content associated with hemodialysis in patients with severe azotemia remains obscure. To investigate whether either interstitial or cytotoxic edema is responsible for dialysis disequilibrium syndrome (DDS), we used diffusion-weighted MR imaging (DWI) to measure the apparent diffusion coefficient (ADC), which is sensitive for detecting tissue water dynamics. METHODS: Eight consecutive patients with end stage renal disease (ESRD) and blood urea nitrogen level of more than 100 mg/dL (160.9 +/- 53.1 mg/dL) were recruited. Conventional MR images, DWI, and clinical manifestations were obtained before and after the 1st hemodialysis. The ADC values were determined for regions of normal-appearing gray and white matter and for regions of hyperintensity of white matter on T2-weighted MR imaging. RESULTS: Foci of bright areas of white matter were found in all patients on T2-weighted images. The ADC values of the patients with ESRD, in white matter and gray matter before and after hemodialysis, were greater than those of the healthy controls (P < .005). Regarding the impact of hemodialysis, the ADC of frontal lobe white matter increased significantly after hemodialysis (1.09 +/- 0.11 versus 1.03 +/- 0.11, P = .036). We did not find the specific area of brain edema reported in posterior leukoencephalopathy and the osmotic demyelination syndrome. CONCLUSIONS: These results suggest that severe azotemia in end stage renal disease leads to interstitial brain edema reflected as increased ADC, and the further increased ADC reflects that edema associated with 1st hemodialysis is interstitial rather than cytotoxic in nature.

Physiological changes during hemodialysis in patients with intradialysis hypertension.

Intradialysis hypertension is a frustrating complication among hemodialysis (HD) patients. This study was conducted to investigate the physiological changes during intradialytic hypertension. The beat-to-beat continuous heart rate, hematocrit (Hct) changes during HD, serum levels of nitric oxide, plasma levels of catecholamine, renin, endothelin (ET-1), cardiac output (CO), and peripheral vascular resistance (PVR) were measured before and after HD in patients prone to develop intradialysis hypertension (n = 30) and from age, sex-matched control HD subjects (n = 30). It was found that the baseline values of Hct, serum levels of nitric oxide, plasma levels of catecholamine, renin, and ET-1, CO, PVR, and power index (low frequency/high frequency ratios) of heart rate variability were not significantly different between the patients and control subjects. In the hypertension-prone group, the plasma levels of catecholamine, renin, and the serial measurements of power index, did not show significant changes. However, the patients showed a significant elevation of systemic vascular resistance (56.8 +/- 9.2% vs 17.7 +/- 9.5; P < 0.05), ET-1 (510.9 +/- 43.3 vs 276.7 +/- 30.1 pg/ml; P < 0.05) and a significant decrease of nitric oxide (NO)/ET-1 balance (0.018 +/- 0.003 vs 0.034 +/- 0.005; P < 0.05) at the end of HD compared with the control patients. It was found that the physiological changes in intradialysis hypertension patients were characterized by inappropriately increased PVR through mechanisms that did not involve sympathetic stimulation or renin activation but might be related with altered NO/ET-1 balance.

Tuberculosis in patients with end-stage renal disease.

OBJECTIVE: To elucidate the clinical manifestations and risk factors of the mortality rate in uraemic patients with tuberculosis (TB) infection. DESIGN: We retrospectively analysed 62 patients with uraemia and active tuberculosis who were admitted to our hospital from 1990 through 2000. The patients were followed up for 2 years after discharge or until death. RESULTS: There were 43 men and 19 women, with a mean age of 63 +/- 13 years. Extra-pulmonary TB was noted in 51.6%. The peritoneum and pleura were the two most common organs involved. Fever of unknown origin was the most common manifestation (77.4%). The corrected serum Ca2+ level of the patients was >10.5 mg/dl in 46.8%. C-reactive protein >6 mg/dl and leukocytosis (white blood cell count >10,000/mm3) at presentation were noted in more than half of the patients. A reversed serum albumin/globulin ratio and leukocytosis were found to be associated with mortality rate. CONCLUSION: More than half of the TB infections in patients with end-stage renal disease presented with extra-pulmonary involvement. Fever of unknown origin, reversed serum albumin/globulin ratio, and unexplained hypercalcaemia in maintenance dialysis patients suggested the possibility of tuberculosis.

[The detection of cyanobacterial cells by a non-fluorescent in situ hybridization method]. / Primenenie metoda gibridizatsii in situ dlia vyiavleniia tsianobakterial'nykh kletok.

Seven cyanobacterial strains (Anabaena macrospora NIER10016, Oscillatoria sp. NIER10042, Microcystis aeruginosa NIER10015, M. ichtyoblabe NIER10025 and NIER10040, M. novacekii NIER10029, and M. wesenbergii NIER10068) were tested by a non-fluorescent in situ hybridization method using two specific horseradish peroxidase--labeled oligonucleotide probes and two chromogenic substrates. This approach was shown to be appropriate for analysis of natural samples.

Acute oxalate nephropathy induced by star fruit in rats.

In this study, we intend to establish a connection between star fruit and acute oxalate nephropathy and also investigate predisposing factors for its development. Male Sprague-Dawley rats of 180 to 200 g were assigned to four groups; namely, control, experimental, fasting, and water-deprivation groups. The former two groups were subjected to both fasting and water deprivation, whereas the latter two groups were subjected to either fasting or water deprivation, respectively. Except for tap water for controls, the remaining groups were administered 4 mL/100 g of body weight of sour star fruit juice with an oxalate concentration of 2.46 g/dL. After these procedures, serial measurement of serum creatinine levels and kidney pathological examination were performed. Peak serum creatinine levels in the control, experimental, fasting, and water-deprivation groups were 0.50 +/- 0.04, 1.46 +/- 0.26, 0.68 +/- 0.20, and 0.52 +/- 0.08 mg/dL, respectively. The experimental group had a greater peak serum creatinine level (P < 0.05). Mean serum creatinine levels of the experimental group days 0, 1, 2, 3, 4, and 5 were 0.43 +/- 0.03, 1.11 +/- 0.18, 1.31 +/- 0.27, 1.16 +/- 0.28, 0.8 +/- 0.26, and 0.82 +/- 0.28 mg/dL, respectively. Mean serum creatinine levels days 1 to 3 were greater than that day 0 (P < 0.05). Pearson's correlation analysis of peak serum creatinine level and kidney weight for the experimental group showed a significant correlation (R = 0.75; P < 0.05; n = 9). In addition to typical changes of oxalate nephropathy, kidney pathological examination showed many refractile oxalate crystals with all rainbow colors under polarized light microscopy in the experimental group. In conclusion, sour star fruit with abundant oxalate contents could cause acute oxalate nephropathy in rats under the conditions of fasting and water deprivation.

Rhabdomyolysis: an unusual feature with mushroom poisoning.

Rhabdomyolysis resulting from mushroom poisoning previously has been unreported in the literature. We present an outbreak of Russula subnigricans poisoning with rhabdomyolysis. The most severely ill patient presented with rhabdomyolysis, severe electrolyte disturbance (hyperkalemia, hypocalcemia), respiratory failure, acute renal failure, pulmonary edema, ventricular tachycardia, and circulatory shock. Mycotoxin may be the cause of rhabdomyolysis. In areas where mushroom gathering is common, mushroom poisoning should be included in the differential diagnosis of rhabdomyolysis.

The long-term effects of single peritonitis episodes on peritoneal equilibration test results in continuous ambulatory peritoneal dialysis patients.

The present study evaluates the long-term effects of single peritonitis episodes on peritoneal equilibration test (PET) results in continuous ambulatory peritoneal dialysis (CAPD) patients. Twenty-five patients (10 men, 15 women) with a mean age of 37.4 +/- 18.7 years were enrolled in this study because all had uneventful peritoneal dialysis periods for more than one year after a first peritonitis episode. Data from a total of 69 PETs were available [25 from before the first peritonitis episode (initial PET), 23 within 1 year after the episode (1-year PET), and 21 within 1-2 years after the episode (2-year PET)]. The changes in the PET results were evaluated using the dialysate-to-plasma ratio of creatinine (D/PCr) and the dialysate-to-instilled glucose ratio (D4/D0) after a 4-hour dwell. The mean values of D/PCr and D4/D0 showed no statistically significant changes between the initial PETs, 1-year PETs, and 2-year PETs. However, analysis of the results for patients with culture-positive peritonitis (n = 17) revealed significant changes in the mean values of D/PCr and D4/D0 between the initial PET and the 2-year PET (0.63 +/- 0.06 vs 0.70 +/- 0.09, p = 0.01; and 0.41 +/- 0.05 vs 0.37 +/- 0.06, p = 0.04, respectively). The long-term effect of a definite bacterial peritonitis episode seems to be an increase in small-molecule transport. In patients with culture-negative peritonitis, the episode had less impact on peritoneal transport.

Establishment of an animal embryo culture system containing various embryotropins and its efficacy for culturing ICR mouse one-cell embryos derived in vivo or in vitro.

OBJECTIVE: To develop an effective ICR mouse embryo culture medium. DESIGN: In vitro model study. SETTING: University-affiliated hospital. ANIMALS: Four-week-old, superovulated mice. INTERVENTION(S): In vivo- or in vitro-derived one-cell embryos were cultured in preimplantation-1 medium (P-1). MAIN OUTCOME MEASURE(S): Preimplantation development. RESULT(S): In vivo-derived embryos were cultured in BSA-containing P-1, to which one of the following substances was added: [1] no addition, [2] amino acids (aa), [3] aa+hemoglobin (hb), [4] aa+hb+cysteine (cys), [5] aa+hb and glucose (glu) added at the four-cell, or [6] aa+hb and glu+cys added at the four-cell stage. More (P<0.05) blastocysts developed after aa or aa+hb addition than after no addition, and glu addition to such medium further stimulated the formation (54%). In P-1 with aa+glu, the addition of 1 microg/mL hb was optimal. Additional improvement of blastocyst formation (78%) was achieved by ethylenediaminetetraacetic acid (EDTA), supplementation and bovine serum albumin replacement with polyvinyl alcohol (PVA) did not inhibit the development. P-1 supplemented with aa, hb, glu, EDTA, and PVA also supported the development of in vitro-derived embryos (70%). CONCLUSION(S): A modified P-1 medium was developed, and it supported the development of both in vivo- and in vitro-derived ICR mouse embryos.

Expression of the Xenopus homologue of the receptor for activated C-kinase 1 (RACK1) in the Xenopus embryo.

We have isolated the Xenopus homologue of the receptor for activated C-kinase 1 (RACK1), whose amino acid sequence shows significant similarity with other vertebrate RACK1s. XRACK1 is a maternally expressed gene and its zygotic expression is detected in the antero-dorsal region and dorsal midline in the late neurula. At tailbud stage, rather diffuse staining is seen in the somite and head. Later, XRACK1 mRNA is expressed highly in ventrally migrating abdominal muscle anlagen, where it remains expressed during subsequent stages.

Increased expression and localization of a serine protease inhibitor, protease nexin-1 (PN-1), in the ovary and uterus during implantation in rat.

Protease nexin-1 (PN-1) is a serine protease inhibitor (serpin) that inactivates several proteases, including thrombin, urokinase, plasminogen activators (PA), and plasmin. It also plays a role in regulating proteolytic activity generated by PA system. PN-1 is known to be involved in tissue remodeling, cellular invasiveness, matrix degradation, and tumor growth. However, the role of PN-1 in female reproductive tracts, such as the uterus, ovary, and oviduct, during pregnancy is not known. The present study was designed to investigate the changes of PN-1 mRNA level and localization in the tracts during implantation and early pregnancy by using reverse transcription (RT)-polymerase chain reaction (PCR) and in situ hybridization. We found that PN-1 mRNA levels were coordinately regulated during early pregnancy in a stage- and tissue-specific manner, such that an increased expression of PN-1 gene appeared at the time of the implantation period in the uterus and ovary. Both the uterus and ovary synthesized PN-1 mRNA and their maximal PN-1 expression occurred on Day 6.5 postcoitum (p.c.). On 13.5 days of pregnancy, PN-1 level was low in the uterus and ovary. On the other hand, PN-1 mRNA in the oviduct did not show after 6.5 days of pregnancy. It appears that PN-1 mRNA in the uterus and ovary was highly regulated during early pregnancy, which might have an important role in implantation of rat blastocysts. PN-1 was localized in endometrial stromal cells of the uterus and in granulosa cells of the unstimulated primary follicles in the ovary during periimplantation period. Also, PN-1 mRNA expression was higher at implantation period than that at nonimplantation period of pregnancy. In conclusion, PN-1 is expressed in female reproductive tracts and highly regulated during implantation and early pregnancy.

Cryopreservation of ICR mouse oocytes: improved post-thawed preimplantation development after vitrification using Taxol, a cytoskeleton stabilizer.

OBJECTIVE: To establish an effective cryopreservation method. DESIGN: In vitro model study. SETTING: Infertility Medical Center, Pochon CHA University. ANIMAL(S): Four-week-old ICR mice superovulated with pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin. INTERVENTION(S): Vitrified-thawed oocytes were fertilized and subsequently cultured in vitro. MAIN OUTCOME MEASURE(S): Post-thawed development, chromosome/spindle normalities, and blastocyst quality. RESULT(S): More cumulus-enclosed oocytes were fertilized and developed to the 8-cell stage after vitrification and thawing than denuded oocytes. However, cryopreserved oocytes of both types had lower spindle and chromosome normalities than fresh oocytes, which resulted in reduced developmental competence after thawing. The addition of 1 microM of Taxol, a cytoskeleton stabilizer, to vitrification solution greatly promoted the blastocyst formation of vitrified-thawed oocytes, compared with no addition (24.0% vs. 58.6%). No difference in blastocyst quality, which was evaluated by blastomere and inner cell mass cell numbers and inner cell mass cell per trophoblast ratio, was found between fresh oocytes and oocytes vitrified with Taxol. CONCLUSION(S): A vitrification solution consisting of 5.5 M ethylene glycol, 1.0 M sucrose, 10% fetal bovine serum, and 1 microM Taxol greatly improved post-thawed development of vitrified oocytes.