In vitro and in vivo efficacy of drugs against the protozoan parasite Azumiobodo hoyamushi that causes soft tunic syndrome in the edible ascidian Halocynthia roretzi (Drasche).

It was discovered recently that infection by a protozoan parasite, Azumiobodo hoyamushi, is the most probable cause for soft tunic syndrome in an edible ascidian, Halocynthia roretzi (Drasche). In an attempt to develop measures to eradicate the causative parasite, various drugs were tested for efficacy in vitro and in vivo. Of the 20 antiprotozoal drugs having different action mechanisms, five were found potent (24-h EC50  < 10 mg L(-1) ) in their parasite-killing effects: formalin, H2 O2 , bithionol, ClO2 and bronopol. Moderately potent drugs (10 < 24-h EC50  < 100 mg L(-1) ) were quinine, fumagillin, amphotericin B, ketoconazole, povidone-iodine, chloramine-T and benzalkonium chloride. Seven compounds, metronidazole, albendazole, paromomycin, nalidixic acid, sulfamonomethoxine, KMnO4 , potassium monopersulphate and citric acid, exhibited EC50  > 100 mg L(-1) . When ascidians were artificially infected with A. hoyamushi, treated using 40 mg L(-1) formalin, bronopol, ClO2 , or H2 O2 for 1 h and then monitored for 24 h, very low mortality was observed. However, the number of surviving parasite cells in the ascidian tunic tissues was significantly reduced by treating with 40 mg L(-1) formalin or ClO2 for 1 h. The data suggest that we might be able to develop a disinfection measure using a treatment regimen involving commonly available drugs.

Elastin-like polypeptide matrices for enhancing adeno-associated virus-mediated gene delivery to human neural stem cells.

The successful development of efficient and safe gene delivery vectors continues to be a major obstacle to gene delivery in stem cells. In this study, we have developed an elastin-like polypeptide (ELP)-mediated adeno-associated virus (AAV) delivery system for transducing fibroblasts and human neural stem cells (hNSCs). AAVs have significant promise as therapeutic vectors because of their safety and potential for use in gene targeting in stem cell research. ELP has been recently employed as a biologically inspired 'smart' biomaterial that exhibits an inverse temperature phase transition, thereby demonstrating promise as a novel drug carrier. The ELP that was investigated in this study was composed of a repetitive penta-peptide with [Val-Pro-Gly-Val-Gly]. A novel AAV variant, AAV r3.45, which was previously engineered by directed evolution to enhance transduction in rat NSCs, was nonspecifically immobilized onto ELPs that were adsorbed beforehand on a tissue culture polystyrene surface (TCPS). The presence of different ELP quantities on the TCPS led to variations in surface morphology, roughness and wettability, which were ultimately key factors in the modulation of cellular transduction. Importantly, with substantially reduced viral quantities compared with bolus delivery, ELP-mediated AAV delivery significantly enhanced delivery efficiency in fibroblasts and hNSCs, which have great potential for use in tissue engineering applications and neurodegenerative disorder treatments, respectively. The enhancement of cellular transduction in stem cells, as well as the feasibility of ELPs for utilization in three-dimensional scaffolds, will contribute to the advancement of gene therapy for stem cell research and tissue regenerative medicine.

Segregation of human neural stem cells in the developing primate forebrain.

Many central nervous system regions at all stages of life contain neural stem cells (NSCs). We explored how these disparate NSC pools might emerge. A traceable clone of human NSCs was implanted intraventricularly to allow its integration into cerebral germinal zones of Old World monkey fetuses. The NSCs distributed into two subpopulations: One contributed to corticogenesis by migrating along radial glia to temporally appropriate layers of the cortical plate and differentiating into lamina-appropriate neurons or glia; the other remained undifferentiated and contributed to a secondary germinal zone (the subventricular zone) with occasional members interspersed throughout brain parenchyma. An early neurogenetic program allocates the progeny of NSCs either immediately for organogenesis or to undifferentiated pools for later use in the "postdevelopmental" brain.

Scrub typhus encephalomyelitis with prominent focal neurologic signs.

BACKGROUND: Encephalomyelitis with prominent focal neurologic signs and associated neuroradiologic abnormalities has not been previously described in scrub typhus. CASE DESCRIPTION: A 22-year-old woman was admitted because of fever and an altered mental state. Neurologic examination revealed bilateral sixth and seventh nerve palsies, bilateral gaze evoked nystagmus, anarthria, dysphagia, quadriparesis, and sensory level at T1. Serum and cerebrospinal fluid samples were positive for tsutsugamushi antibody. The patient's magnetic resonance images demonstrated the lesions responsible for the neurologic dysfunctions: in the lower brainstem, cerebellar peduncles, and spinal cord. It was interesting that the gray matter of the spinal cord was predominantly involved. CONCLUSIONS: The recognition of unusual manifestations and the clinical suspicion of this treatment-responsive disease may be important, particularly in the face of increasing international and intranational travel.

Neural stem cells are uniquely suited for cell replacement and gene therapy in the CNS.

In recent years, it has become evident that the developing and even the adult mammalian CNS contain a population of undifferentiated, multipotent cell precursors, neural stem cells, the plastic properties of which might be of advantage for the design of more effective therapies for many neurological diseases. This article reviews the recent progress in establishing rodent and human clonal neural stem cell lines, their biological properties, and how these cells can be utilized to correct a variety of defects, with prospects for the near future to harness their behaviour for neural stem cell-based treatment of diseases in humans.

Transplantation of neural progenitor and stem cells: developmental insights may suggest new therapies for spinal cord and other CNS dysfunction.

Multipotent neural progenitors and stem cells may integrate appropriately into the developing and degenerating central nervous system. They may also be effective in the replacement of genes, cells, and nondiffusible factors in either a widespread or a more circumscribed manner, depending on the therapeutic demands of the clinical situation. In addition, they may be uniquely responsive to some types of neurodegenerative conditions. We believe that these various appealing capabilities are the normal expression of basic biologic properties and attributes of a stem cell. The therapeutic utility of some of those properties is illustrated in this review of ongoing work in our laboratory, particularly with regard to spinal dysfunction. In these examples, we believe we have tapped into a mechanism that underlies a remarkable degree of natural plasticity programmed into the nervous system at the cellular level, and we have now exploited those properties for therapeutic ends.

Characterization of an inducible oxidative stress response in Vitreoscilla C1.

Vitreoscilla becomes resistant to killing by hydrogen peroxide and heat shock when pretreated with nonlethal levels of hydrogen peroxide. The pretreated Vitreoscilla cells (60 microM hydrogen peroxide for 120 min) significantly increased survival of the lethal dose of 20 mM hydrogen peroxide or heat shock (22 degrees C --> 37 degrees C). This indicates the existence of an adaptive response to oxidative stress. However, cells pretreated with 60 microM hydrogen peroxide became nonresistant to a lethal dose of a menadione. This result shows that hydrogen peroxide does not induce cross-resistance to menadione in Vitreoscilla. Furthermore, Vitreoscilla treated with hydrogen peroxide, heat shock, and menadione showed a change in the protein composition, as monitored by a two-dimensional gel analysis. During adaptation to hydrogen peroxide, 12 proteins were induced. Also, 18 new proteins synthesized in response to heat shock were detected by a 2-D gel analysis. The redox-cycling agents also elicited the synthesis of 6 other proteins that were unseen with hydrogen peroxide.

Alleviation of apoptosis by serum in Chinese hamster ovary cells ectopically expressing human Fas antigen.

Fas-mediated apoptosis is an important regulatory mechanism for the development of T-cells and prevention of oncogenesis. Here, we establish Chinese hamster ovary (CHO) cell lines which stably express Fas antigen, and analyzed apoptosis induced by anti-Fas IgM. While Fas-transfected hamster cells did not undergo apoptosis when stimulated with anti-Fas antibody in the presence of medium containing 10% serum, in reduced serum concentrations, anti-Fas antibody caused these cells to round up and detach from the culture dish. Analysis of the DNA content by a flow cytometry demonstrated a significant increase of cells with sub-G1 amount of DNA upon Fas stimulation in the low serum concentrations. The increase in the number of apoptosis cells was inhibited by an apopain (CPP32, caspase 3) inhibitor or insulin-like growth factor-I. In contrast, apoptosis in a Fas-transfected mouse T-cell line occurred in the presence of 10% serum. these results suggest that factors including insulin-like growth factor-I in fetal bovine serum protect CHO cells from apopain-dependent apoptosis mediated by Fas-antigen stimulation.

Transplantation of neural stem cells: cellular & gene therapy for hypoxic-ischemic brain injury.

We have tracked the response of host and transplanted neural progenitors or stem cells to hypoxic-ischemic (HI) brain injury, and explored the therapeutic potential of neural stem cells (NSCs) injected into mice brains subjected to focal HI injury. Such cells may integrace appropriately into the degenerating central nervous system (CNS), and showed robust engraftment and foreign gene expression within the region of HI inury. They appeared to have migrated preferentially to the site of ischemia, experienced limited proliferation, and differentiated into neural cells lost to injury, trying to repopulate the damaged brain area. The transplantation of exogenous NSCs may, in fact, augment a natural self-repair process in which the damaged CNS "attempts" to mobilize its own pool of stem cells. Providing additional NSCs and trophic factors may optimize this response. Therefore, NSCs may provide a novel approach to reconstituting brains damaged by HI brain injury. Preliminary data in animal models of stroke lends support to these hypotheses.

Effect of tolazoline on persistent hypoxemia in severe hyaline membrane disease.

Ten critically-ill preterm infants with severe hyaline membrane disease received tolazoline because of persistent hypoxemia refractory to the administration of 100% oxygen and mechanical ventilation. Seven infants (70%) responded immediately with an increase in PaO2 greater than or equal to 20 mmHg in the umbilical arterial gas within 60 minutes after bolus infusion (1 to 2 mg/kg) of tolazoline. Twenty-four hours later after the tolazoline infusion, the FiO2 had been decreased from 1.0 to a mean of 0.82 +/- 0.16, and the MAP from 16.5 +/- 1.8 to 15.6 +/- 4.5 cm H2O. Four of 7 infants (57%) who had an immediate response survived, whereas none survived out of 3 infants who failed to respond initially. Three infants experienced relatively severe complications possibly related to tolazoline. There appears to be a place for the use of tolazoline in a severely hypoxemic infant with hyaline membrane disease who is being ventilated, and in whom arterial oxygenation cannot be improved by a further increase in the inspired oxygen concentration or by an alteration of ventilator settings.

Evaluation of a simultaneous HIV antigen and antibody detection test in Korean population.

Current diagnosis of human immunodeficiency virus (HIV) infection relies on the detection of anti-HIV antibodies by enzyme-linked immunosorbent assay (ELISA). Recently, kits detecting both p24 antigenemia and anti-HIV/anti-HIV2 antibodies have been developed. Thus, it is necessary to compare those kits developed as such. The aim of this study was to evaluate the diagnostic efficiency of a simultaneous detection test of p24 antigen and anti-HIV1/2 antibodies in a low prevalence area. Eight hundred and four randomly selected sera proven negative for HIV infection and 110 sera from 54 patients diagnosed as HIV infected, obtained between 1999 and 2000, were used for this study. One commercial lot of panels composed of consecutive sera obtained from known HIV-infected patient was included. Anti-HIV1/2 antibodies were detected by two different commercial ELISA kits, one from Korean and the other from German manufacturer. P24 antigen test was performed by ELISA. The simultaneous HIV antigen and antibody detection test was carried out. In the meantime, HIV RNA PCR and anti-HIV and anti-HIV2 western blot assays were also performed to confirm the test results in cases the test results didn't agree. The simultaneous detection kit showed 100% sensitivity and 99.6% specificity. Furthermore, the test displayed the possibility of earlier diagnosis than conventional anti-HIV1/2 ELISA with the results obtained from a group of consecutive panel sera infected with HIV. From these results, we concluded that the simultaneous HIV antigen and antibody detection test can be applied as a substitute clinical screening test in the place of conventional anti-HIV1/2 ELISA, and there is the probable benefit of early diagnosis.

Influence of nutrition on anti-tumor activity.

Nutritionally supporting the malnourished tumor bearing host may not benefit the disease outcome, but, rather, may preferentially "feed the cancer". We hypothesized that repletion is beneficial only when it augments an anti-tumor immune response. To support this hypothesis, 240 A/J mice were assigned to isocaloric dietary groups (24%, 5%, or 2.5% protein). On day 14 the mice received either immunogenic C1300- neuroblastoma (NB) or non-immunizing TBJ-NB. On day 21 half of the restricted animals were repleted with 24% protein chow. At day 35, chromium-release cell-mediated cytotoxicity was measured. In the group of mice that received 2.5% protein chow, nutritional repletion specifically augmented anti-tumor activity for C1300-NB which elicits a host immune response (33.78 L.U. (repleted) vs 3.47 L.U. (depleted) p less than 0.01), in contrast, nutritional repletion was detrimental for non-immunizing TBJ-NB, where further depression of cytotoxicity was seen (1.37 L.U. (repleted) vs 2.06 L.U. (depleted) 0 less than 0.01). This suggests that the influence of nutritional repletion in tumor nearing animals is dependent on the integrity of host's anti-tumor immunity.

Tourette disorder and HLA typing.

HLA A, B, C and DR were typed in 73 Korean patients with Tourette disorder meeting the diagnostic criteria of DSM III-R and compared with 291 normal subjects. Relatively higher frequencies were found in HLA A11 and A26(10) with lower incidences in HLA A24(9) and B13. A family history of tic disorders was associated with a lower frequency of HLA A24(9).

Exogenous surfactant replacement therapy of hyaline membrane disease in premature infants.

We conducted a clinical trial to assess whether surfactant-TA given within the first six hours of life could improve oxygenation and reduce the ventilatory support in premature infants with hyaline membrane disease (HMD) during the first 24 hours of life. Eight premature infants with severe HMD requiring ventilation were treated, at a mean age of 2.72 hours, with a single intratracheal instillation of surfactant-TA (120 mg/kg). Arterial oxygenation improved dramatically as reflected by the increase of the a/A PO2 ratio and PaO2 to about 2 times the pretreatment values within 3 hours after surfactant treatment. And thus, oxygen concentrations (FiO2) could be reduced and remained significantly lower than pretreatment values during the first 24 hours after treatment. Infants given surfactant-TA required lower mean airway pressure (MAP) and had a significantly decreased ventilatory index (VI) during the first 24 hours after treatment, which reflect the decreased requirement for ventilatory support. Chest radiograph scores significantly improved within 24 hours after treatment compared with pretreatment scores. In this trial, we found that a single intratracheal dose of surfactant-TA given to infants with HMD resulted in improved respiratory status and radiographic findings during the first 24 hours after treatment.

Prospective evaluation of perinatal risk factors for cerebral palsy and delayed development in high risk infants.

Prematurity, intrauterine infection and perinatal brain injury have been reported to be significant risk factors of cerebral palsy (CP). We examined the perinatal predictors of cerebral palsy and delayed development (DD) in 184 high risk infants. Thirty-five infants were diagnosed as cerebral palsy and delayed development at 12 months corrected age. Antenatal, intrapartum, and neonatal factors were prospectively evaluated in 2 groups of high risk infants compared with controls; Group A (n = 79), infants weighing less than 2,000 g; Group B (n = 43), infants weighing 2,000 g or more. In univariate analysis, there were no significant antenatal and intrapartum factors associated with cerebral palsy and delayed development in either group. We found that significant postnatal risk factors of CP in group A included sepsis (p = 0.008), BPD (bronchopulmonary dysplasia) (p = 0.028), IVH (intraventricular hemorrhage) (p = 0.042), ventriculomegaly (VM) (p = 0.001) and a longer duration of mechanical ventilation (p = 0.001); while in group B, sepsis (p = 0.047) and neonatal seizure (p = 0.027) were significant risk factors. In multivariate analysis, sepsis in group B was a moderate risk factor of CP (OR (odds ratio) 1.47; 95% CI (confidence interval) 1.02-2.13). In conclusion, neonatal sepsis may contribute to the development of cerebral palsy and delayed development. We suggest that high risk infants who have sepsis should be carefully followed for cerebral palsy and delayed development. The prevention of cerebral palsy may be feasible by decreasing neonatal risk factors such as sepsis during the neonatal period.

[Clinical analysis of bladder cancer patients treated by radical cystectomy].

We assessed the treatment outcome of 105 patients with transitional cell carcinoma of the bladder treated by total cystectomy at our university hospital, between 1979 and 1993. The patients consisted of 84 men and 21 women (male to female ratio : 4:1), between 45 and 82 years old (mean, 65.5 years old). The overall cancer-specific survival rate at 3 and 5 years was 76.3% and 68.9%, respectively. The 5-year survival rate was 85.2% for grade 2 and 59.9% for grade 3 tumors with a significant difference in the survival curves between the two groups (p < 0.05). The 5-year survival rate according to pathological stage was 100% for pTa, 75.6% for pT1, 78.4% for pT2, 54.0% for pT3 and 39.8% for pT4. A significant difference was observed between pTa and pT3 (p < 0.05), and between pTa-2 and pT4 (p < 0.05). The 5-year survival rate was 72.3% for patients without lymph node involvement and 11.9% for those with lymph node involvement, the difference being significant (p < 0.01). Nineteen patients who received pre- and/or post-operative chemotherapy did not show a higher 5-year survival rate than those who did not.

[Adenocarcinoma of the bladder 19 years after the augmentation ileocystoplasty: report of a case].

We report a case of adenocarcinoma of the augmented bladder 19 years after ileocystolasty. The patient was a 53-year-old man who underwent right nephrectomy and ileocystoplasty (Pyrah's method) for contracted bladder due to tuberculosis in 1965. In another hospital, transurethral resection (TUR) was performed against a tumor in the anastomotic site between the bladder and the ileal segment in 1996. Histopathological examination of the specimen obtained by TUR revealed poorly-differentiated mucinous adenocarcinoma. In our hospital, partial cystectomy with total resection of ileal segment and ileocystoplasty were performed. The tumors located in the anastomotic site between the bladder and ileal segment as well as in the ileal segment. Histopathological examination revealed poorly-differentiated mucinous adenocarcinoma. The patient has survived 12 months without any evidence of tumor recurrence. To our knowledge, this is the eighth case report in Japan.

[Tuberous sclerosis associated with renal angiomyolipoma, pulmonary lymphangioleiomyomatosis and subungual fibroma: report of a case].

We report a case of tuberous sclerosis associated with bilateral renal angiomyolipomas (AMLs), pulmonary lymphangioleiomyomatosis (LAM) and subungual fibroma of hands and feet. A 42-year-old woman who was diagnosed as tuberous sclerosis at the age of 18 complained of left flank pain and abdominal fullness. Bilateral renal AMLs were pointed out when complete examinations were performed for hypertension at the age of 32. She suffered from severe left flank pain and abdominal distension due to the left renal tumor. Left nephrectomy and excision of the renal hilar tumor were performed. The left renal tumor weighed 1120 g, the perirenal space was filled with the tumor. histopathological diagnosis of the left renal tumor and renal hilar tumor was AML. In our case, bilateral pneumothorax appeared, and chest CT scan revealed bilateral multiple pulmonary cysts. Histopathological diagnosis of pulmonary cysts was LAM. Other complications of our case are intracranial calcification and adenoma sebaceum.

[Bilateral synchronous multilocular epididymal cysts: a case report].

A rare case of bilateral synchronous multilocular epididymal cysts is reported. A fifty-six year old man visited to our hospital with a chief complaint of swelling of bilateral intrascrotal contents. Ultrasonographic findings demonstrated multilocular lesions of the bilateral intrascrotal contents. Preoperative diagnosis was bilateral multilocular hydrocele testes. Operative procedure revealed bilateral cysts originating from the head, body and tail of the epididymis without the cysts of the tunica vaginalis. Operative sight was bilateral synchronous multilocular epididymal cysts, and bilateral epididymal cystectomy were performed. The specimen size was right diameter 12 x 6 cm and left its 8 x 5.5 cm. The puncture of the cystic fluid revealed many spermatozoa in both sides. The acquired cysts of the epididymis generally are the result of tubular obstruction with dilation of tubules adjacent to the obstruction. The dilated tubules are filled variably with viable and degenerating spermatozoa. The cysts originate most commonly from the head of the epididymis, and are unilateral, unilocular or multilocular and are usually within 1 cm in diameter. In our case, bilateral synchronous epididymal cysts originating from not only the head, but also the body and the tail is a rare case.

[A case of rapidly progressive IgA nephropathy with transient hypocomplementemia at onset].

We present a case of IgA nephropathy (IgAGN) which developed rapidly progressive glomerulonephritis and showed marked clinical improvement with treatment. The patient was a 7-year-old boy who initially presented with acute nephritic syndrome with hypocomplementemia. Although the renal function improved with normalization of the serum complement level, it deteriorated again progressively. The first renal biopsy revealed cellular crescents in about 70 percent of 43 glomeruli. Immunofluorescent microscopy demonstrated deposits of IgA, C3 and IgG in the mesangium; they were also deposited along the glomerular capillary walls. He was treated with plasma exchange associated with hemodialysis and methylprednisolone pulse therapy, followed by oral administration of prednisolone, cyclophosphamide and warfarin. Renal function recovered to the normal range about two months after the initiation of treatment. The second biopsy demonstrated a marked decrease in histological activity. In this case, transient hypocomplementemia at onset may indicate that acute glomerulonephritis caused exacerbation of clinically silent IgAGN. Aggressive therapy may be effective in patients with rapidly progressive IgAGN if treated at an early stage.