The roles of testicular nuclear receptor 4 (TR4) in male fertility-priapism and sexual behavior defects in TR4 knockout mice.

BACKGROUND: Successful reproductive efforts require the establishment of a situation favorable for reproduction that requires integration of both behavior and internal physiological events. TR4 nuclear receptor is known to be involved in male fertility via controlling spermatogenesis, yet its roles in regulating other biological events related to reproduction have not been completely revealed. METHODS: Male TR4 knockout (TR4 -/-) and wild type mice were used for the sexual behavior and penile dysfunction studies. Mice were sacrificed for histological examination and corresponding genes profiles were analyzed by quantitative RT-PCR. Reporter gene assays were performed. RESULTS: We describe an unexpected finding of priapism in TR4 -/- mice. As a transcriptional factor, we demonstrated that TR4 transcriptionally modulates a key enzyme regulating penis erection and neuronal nitric oxide synthese NOS (nNOS). Thereby, elimination of TR4 results in nNOS reduction in both mRNA and protein levels, consequently may lead to erectile dysfunction. In addition, male TR4 -/- mice display defects in sexual and social behavior, with increased fear or anxiety, as well as reduced mounting, intromission, and ejaculation. Reduction of ER alpha, ER beta, and oxytocin in the hypothalamus may contribute to defects in sexual behavior and stress response. CONCLUSIONS: Together, these results provide in vivo evidence of important TR4 roles in penile physiology, as well as in male sexual behavior. In conjunction with previous finding, TR4 represents a key factor that controls male fertility via regulating behavior and internal physiological events.

Mice lacking TR4 nuclear receptor develop mitochondrial myopathy with deficiency in complex I.

The estimated incidence of mitochondrial diseases in humans is approximately 1:5000 to 1:10,000, whereas the molecular mechanisms for more than 50% of human mitochondrial disease cases still remain unclear. Here we report that mice lacking testicular nuclear receptor 4 (TR4(-/-)) suffered mitochondrial myopathy, and histological examination of TR4(-/-) soleus muscle revealed abnormal mitochondrial accumulation. In addition, increased serum lactate levels, decreased mitochondrial ATP production, and decreased electron transport chain complex I activity were found in TR4(-/-) mice. Restoration of TR4 into TR4(-/-) myoblasts rescued mitochondrial ATP generation capacity and complex I activity. Further real-time PCR quantification and promoter studies found TR4 could modulate complex I activity via transcriptionally regulating the complex I assembly factor NDUFAF1, and restoration of NDUFAF1 level in TR4(-/-) myoblasts increased mitochondrial ATP generation capacity and complex I activity. Together, these results suggest that TR4 plays vital roles in mitochondrial function, which may help us to better understand the pathogenesis of mitochondrial myopathy, and targeting TR4 via its ligands/activators may allow us to develop better therapeutic approaches.

Abnormal inner root sheath of the hair follicle in the loose anagen hair syndrome: an ultrastructural study.

BACKGROUND: Loose anagen hair syndrome (LAHS) is a disorder in which the hair pulls out easily and painlessly from the scalp. It first manifests in early childhood when the main concern of parents is that the sparse hair does not grow. The hair density and length improve with age, but the looseness persists into adulthood. OBJECTIVE: Light and electron microscopic studies of hair follicles were performed to better define the microscopic changes seen in LAHS. METHODS: Biopsy specimens were obtained from 4 patients, 3 children and 1 adult. The hair follicles were studied by light and electron microscopy. RESULTS: The most conspicuous structural changes were found in the inner root sheath complex of the anagen follicle. With light microscopy, the keratinized Henle cell layer showed a tortuous and irregular swelling. Irregular keratinization of the cuticle cells of the inner root sheath and a swollen appearance of Huxley cells were also found. With electron microscopy, the major pathological changes consisted of intercellular edema in the prekeratinized Huxley cell zone and dyskeratosis of Henle cells and cuticle cells of the inner root sheath. LIMITATIONS: The studies were done on a small number of patients. CONCLUSIONS: Structural abnormalities of the inner root sheath appear to disturb its normal supportive and anchoring function and result in a loose attachment of the hair shaft to the anagen follicle.

Abnormal cerebellar cytoarchitecture and impaired inhibitory signaling in adult mice lacking TR4 orphan nuclear receptor.

Since testicular orphan nuclear receptor 4 (TR4) was cloned, its physiological functions remain largely unknown. In this study, the TR4 knockout (TR4(-/-)) mouse model was used to investigate the role of TR4 in the adult cerebellum. Behaviorally, these null mice exhibit unsteady gait, as well as involuntary postural and kinetic movements, indicating a disturbance of cerebellar function. In the TR4(-/-) brain, cerebellar restricted hypoplasia is severe and cerebellar vermal lobules VI and VII are underdeveloped, while no structural alterations in the cerebral cortex are observed. Histological analysis of the TR4(-/-) cerebellar cortex reveals reductions in granule cell density, as well as a decreased number of parallel fiber boutons that are enlarged in size. Further analyses reveal that the levels of GABA and GAD are decreased in both Purkinje cells and interneurons of the TR4(-/-) cerebellum, suggesting that the inhibitory circuits signaling within and from the cerebellum may be perturbed. In addition, in the TR4(-/-) cerebellum, immunoreactivity of GluR2/3 was reduced in Purkinje cells, but increased in the deep cerebellar nuclei. Together, these results suggest that the behavioral phenotype of TR4(-/-) mice may result from disrupted inhibitory pathways in the cerebellum. No progressive atrophy was observed at various adult stages in the TR4(-/-) brain, therefore the disturbances most likely originate from a failure to establish proper connections between principal neurons in the cerebellum during development.

Impact of intensive-care-unit(ICU)-acquired ventilator-associated pneumonia(VAP) on hospital mortality: a matched-paired case-control study.

The effect of ICU-acquired ventilator-associated pneumonia (VAP) on hospital mortality is still a controversial issue in many countries. The aim of this study was to evaluate the effect of ICU-acquired VAP on hospital mortality in a Japanese university hospital. Our study population was comprised of patients aged 16 years or older who were admitted to our ICU and received mechanical ventilation for more than 48 hours during a period of 42 months as of December 2003. To evaluate whether VAP was an independent risk factor for hospital mortality after controlling for other clinical factors, patients with fatal outcomes (cases) were compared to those who survived (controls). From 587 eligible patients, we analyzed 75 cases and 150 controls who were successfully matched on sex, age, and the Acute Physiology and Chronic Health Evaluation II (APACHE II) score using conditional logistic regression models. Univariate analysis demonstrated that hemodialysis (odds ratio [OR], 2.24; 95% confidence interval [CI], 1.21-4.15; p = 0.01), surgical site infection (OR, 2.45; 95% CI, 1.22-4.91; p = 0.01), and VAP (OR, 2.69; 95% CI, 1.55-4.69; p < 0.001) were significantly associated with hospital mortality. After adjusting for confounding factors, multivariate conditional logistic regression analysis showed that hemodialysis (OR, 2.05; 95% CI, 1.06-3.94; p = 0.03) and VAP (OR, 2.20; 95% CI, 1.10-4.39; p = 0.03) were independently associated with hospital mortality. In conclusion, these data suggest that ICU-acquired VAP significantly affects hospital mortality.

Incidence and outcomes of ventilator-associated pneumonia in Japanese intensive care units: the Japanese nosocomial infection surveillance system.

OBJECTIVES: To determine the incidence of ventilator-associated pneumonia (VAP) among intensive care unit (ICU) patients in Japan and to assess the impact of VAP on patient outcomes, including mortality, length of stay, and duration of mechanical ventilation. DESIGN: Multicenter cohort study. SETTING: Twenty-eight ICUs in multidisciplinary Japanese hospitals with more than 200 beds. PATIENTS: A total of 21,909 patients 16 years or older who were admitted to an ICU between June 2002 and June 2004, stayed in the ICU for 24 to 1,000 hours, and were not transferred to another ICU. RESULTS: The overall infection rates for nosocomial pneumonia and VAP were 6.5 cases per 1,000 patient-days and 12.6 cases per 1,000 ventilator-days, respectively. The standardized mortality rates for the patients with VAP was 1.3 (95% confidence interval [CI], 1.1-1.6): 1.1 (95% CI, 0.9-1.4) for the cases due to drug-susceptible pathogens and 1.5 (95% CI, 1.1-1.9) for the cases due to drug-resistant pathogens. After adjusting for Acute Physiology and Chronic Health Evaluation II score, the mean length of stay for the patients with VAP caused by drug-susceptible pathogens (15.2 days [95% CI, 14.6-15.8]) and by drug-resistant pathogens (17.8 days [95% CI, 17.0-18.6]) was significantly longer than that in the patients without nosocomial infection (6.8 days [95% CI, 6.7-6.9]). The mean duration of mechanical ventilation in the patients with VAP caused by drug-susceptible pathogens (12.0 days [95% CI, 11.5-12.5]) and drug-resistant pathogens (14.1 days [95% CI, 13.5-14.8]) was significantly longer than that in the patients without nosocomial infection (4.7 days [95% CI, 4.6-4.8]). CONCLUSION: The incidence of VAP is substantial among ICU patients in Japan. The potential impact of VAP on patient outcomes emphasizes the importance of preventive measures against VAP, especially for VAP caused by drug-resistant pathogens.

Deficits in motor coordination with aberrant cerebellar development in mice lacking testicular orphan nuclear receptor 4.

Since testicular orphan nuclear receptor 4 (TR4) was cloned, its physiological function has remained largely unknown. Throughout postnatal development, TR4-knockout (TR4-/-) mice exhibited behavioral deficits in motor coordination, suggesting impaired cerebellar function. Histological examination of the postnatal TR4-/- cerebellum revealed gross abnormalities in foliation; specifically, lobule VII in the anterior vermis was missing. Further analyses demonstrated that the laminations of the TR4-/- cerebellar cortex were changed, including reductions in the thickness of the molecular layer and the internal granule layer, as well as delayed disappearance of the external granule cell layer (EGL). These lamination irregularities may result from interference with granule cell proliferation within the EGL, delayed inward migration of postmitotic granule cells, and a higher incidence of apoptotis. In addition, abnormal development of Purkinje cells was observed in the postnatal TR4-/- cerebellum, as evidenced by aberrant dendritic arborization and reduced calbindin staining intensity. Expression of Pax-6, Sonic Hedgehog (Shh), astrotactin (Astn), reelin, and Cdk-5, genes correlated with the morphological development of the cerebellum, is reduced in the developing TR4-/- cerebellum. Together, our findings suggest that TR4 is required for normal cerebellar development.

Growth retardation and abnormal maternal behavior in mice lacking testicular orphan nuclear receptor 4.

Testicular orphan nuclear receptor 4 (TR4) is a member of the nuclear receptor superfamily for which a ligand has not yet been found. In vitro data obtained from various cell lines suggest that TR4 functions as a master regulator to modulate many signaling pathways, yet the in vivo physiological roles of TR4 remain unclear. Here, we report the generation of mice lacking TR4 by means of targeted gene disruption (TR4(-/-)). The number of TR4(-/-) pups generated by the mating of TR4(+/-) mice is well under that predicted by the normal Mendelian ratio, and TR4(-/-) mice demonstrate high rates of early postnatal mortality, as well as significant growth retardation. Additionally, TR4(-/-) females show defects in reproduction and maternal behavior, with pups of TR4(-/-) dams dying soon after birth with no indication of milk intake. These results provide in vivo evidence that TR4 plays important roles in growth, embryonic and early postnatal pup survival, female reproductive function, and maternal behavior.

[Domestic violence--public health perspectives].

Domestic violence (DV) implies violence against women by intimate male partners. DV is a serious health issue for women, as well as a violation of human rights. It is a challenge to develop effective public health interventions, as they have to take into account complicated social and psychological background factors. In this paper we present an overview of various interventions in Japan and elsewhere in the world up to now, and propose a strategy for developing effective public health interventions. Governments and NGOs have been involved in various interventions to eliminate DV, e.g., establishing legal frameworks, providing emergency shelters for abused women, and educating male abusers. Health sector interventions include: systematic DV education to health professionals in Europe and the United States; and development of DV victim support networks, in which health facilities play core roles, in Asia and Latin America. The major expected roles of health professionals are identification and treatment of abused women, and prevention of recurrent violence. However, achievement of those goals is insufficient, because of the lack of systematic education, different views on DV between health professionals and abused women, misunderstanding of background factors, and lack of coordination between relevant agencies. The health sector, including clinical and public health services, is expected to play important roles in identifying and supporting abused women in Japan. A possible strategy is to integrate DV interventions into existing maternal and child health service systems. All the front-line health professionals should be provided with systematic training and practical manuals to treat abused women. Further research and evaluation of past interventions are needed to develop effective interventions.

Time profile of cerebral [18F]6-fluoro-L-DOPA metabolites in nonhuman primate: implications for the kinetics of therapeutic L-DOPA.

At least two rates of dopamine turnover have been demonstrated in vivo, including a slow turnover rate that is associated with synaptic vesicles, and a faster rate that leads to rapid production of dopamine metabolites. Similarly, [18F]6-fluorodopamine (FDA), the decarboxylation product of the PET tracer [18F]6-fluoro-L-DOPA (FDOPA), may have multiple turnover rates which could substantially affect the interpretation of FDOPA uptake. To better characterize FDA turnover in vivo, we measured the formation of FDOPA metabolites in primate brain following bolus FDOPA injection with carbidopa pretreatment. FDOPA was allowed to circulate for either 30 minutes or 90 minutes, prior to removal of brain samples. The primary metabolites in striatum were [18F]6-fluoro-3-methyl-L-DOPA (3-OMFD), FDA, [18F]6-fluoro- L-3,4-dihydroxyphenylacetic acid (FDOPAC), and [18F]6-fluorohomovanillic acid (FHVA). The percentages of total radioactivity in striatum at 30 minutes and 90 minutes were: FDOPA(5%, 2%), FDA (39%, 23%), FDOPAC (12%, 3%), FHVA (14%, 34%), and 3-OMFD (29%, 39%). In cortex and cerebellum most of the activity (73%, 80%) was 3-OMFD. These data were compared against the metabolite profiles predicted by two compartmental models of FDOPA metabolism. A model that assumes only a single slow rate of FDA turnover predicted much lower concentrations of FDA metabolites (FDOPAC, FHVA) in striatum than were found in the brain assay, while a model that includes both slow and fast FDA turnover was in much better agreement. These findings extend and confirm previous observations of FDOPA metabolites. The implications for the interpretation of FDOPA PET, particularly in terms of the availability of dopamine synthesized from therapeutic L-DOPA, are discussed.

Effect of latanoprost on hair growth in the bald scalp of the stump-tailed macacque: a pilot study.

Latanoprost, a selective FP prostanoid receptor agonist used in the treatment of glaucoma, has a hypertrichotic side effect. Using the macaque model of androgenetic alopecia, we examined the effect of latanoprost on hair growth. Eight monkeys were divided into 2 groups; one group received a daily topical application of 50 microg/ml of latanoprost for 5 months; a control group had a daily application of vehicle. For an additional 3 months, 2 monkeys from each group were given 500 microg/ml latanoprost, while the remaining monkeys continued with the previous treatment. Hair growth was evaluated by monthly photographs and phototricho-graphic analysis. Fifty microg/ml of latanoprost caused minimal hair growth. Latanoprost at 500 microg/ml induced moderate to marked hair regrowth with 5-10% conversion of vellus hairs to intermediary or terminal hairs. The vehicle group showed no effect. Further evaluation of latanoprost as an agent for treatment of human androgenetic alopecia is indicated.