Characterization of pathological stages in a mouse model of progressive multiple sclerosis.

The purpose of this study was to analyze and elucidate the mechanisms of non-obese diabetes-experimental autoimmune encephalomyelitis (NOD-EAE), an animal model of progressive multiple sclerosis (MS), and to compare the pathological features with those observed in human progressive MS. Pathological analysis, flow cytometry analysis, immunohistochemical staining, and transcriptome analysis were performed at each pathological stage of the NOD-EAE mice to characterize each pathological stage in the lesion. The NOD-EAE mice showed a biphasic pattern of disease progression once in remission. The longitudinal profile of demyelination and inflammatory cell infiltration in the spinal cord was consistent with the pathological score. In the chronic phase of the disease, fibrosis and lymph follicle formation, characteristic of progressive human MS, were observed. Here we describe the pathological profile and transcriptome analysis of the NOD-EAE mice and verify that this model has similar features to those of human progressive MS. Our findings suggest that this model recapitulates lymph follicle formation, a disease hallmark of progressive MS, and fibrosis, a feature complicating the pathogenesis of MS in the chronic phase. This model may be useful for evaluating the efficacy of therapeutic agents and for mechanistic analysis.

Distinct Mechanism of Cysteine Oxidation-Dependent Activation and Cold Sensitization of Human Transient Receptor Potential Ankyrin 1 Channel by High and Low Oxaliplatin.

Oxaliplatin, a third-generation platinum-based chemotherapeutic agent, displays unique acute peripheral neuropathy triggered or enhanced by cold, and accumulating evidence suggests that transient receptor potential ankyrin 1 (TRPA1) is responsible. TRPA1 is activated by oxaliplatin via a glutathione-sensitive mechanism. However, oxaliplatin interrupts hydroxylation of a proline residue located in the N-terminal region of TRPA1 via inhibition of prolyl hydroxylase (PHD), which causes sensitization of TRPA1 to reactive oxygen species (ROS). Furthermore, PHD inhibition endows cold-insensitive human TRPA1 (hTRPA1) with ROS-dependent cold sensitivity. Since cysteine oxidation and proline hydroxylation regulate its activity, their association with oxaliplatin-induced TRPA1 activation and acquirement of cold sensitivity were investigated in the present study. A high concentration of oxaliplatin (1 mM) induced outward-rectifier whole-cell currents and increased the intracellular Ca2+ concentration in hTRPA1-expressing HEK293 cells, but did not increase the probability of hTRPA1 channel opening in the inside-out configuration. Oxaliplatin also induced the rapid generation of hydrogen peroxide, and the resultant Ca2+ influx was prevented in the presence of glutathione and in cysteine-mutated hTRPA1 (Cys641Ser)-expressing cells, whereas proline-mutated hTRPA1 (Pro394Ala)-expressing cells showed similar whole-cell currents and Ca2+ influx. By contrast, a lower concentration of oxaliplatin (100 µM) did not increase the intracellular Ca2+ concentration but did confer cold sensitivity on hTRPA1-expressing cells, and this was inhibited by PHD2 co-overexpression. Cold sensitivity was abolished by the mitochondria-targeting ROS scavenger mitoTEMPO and was minimal in cysteine-mutated hTRPA1 (Cys641Ser or Cys665Ser)-expressing cells. Thus, high oxaliplatin evokes ROS-mediated cysteine oxidation-dependent hTRPA1 activation independent of PHD activity, while a lower concentration induces cold-induced cysteine oxidation-dependent opening of hTRPA1 via PHD inhibition.

Simultaneous measurement of cerebral hemoglobin oxygen saturation and blood volume in asphyxiated neonates by near-infrared time-resolved spectroscopy.

BACKGROUND: Hypoxic-ischemic encephalopathy (HIE) usually results in a poor clinical outcome even when treated with hypothermic therapy (HT). Early postnatal changes in cerebral blood oxygenation and hemodynamics may be critical determinants of brain injury and the efficacy of HT. OBJECTIVES: We measured cerebral hemoglobin oxygen saturation (ScO2) and cerebral blood volume (CBV) by near-infrared time-resolved spectroscopy (TRS) in HT-treated and non-HT-treated neonatal HIE patients to assess the influence of these parameters on clinical outcome. METHODS: We retrospectively compared ScO2, CBV, and clinical outcomes of 11 neonates with HIE: 5 were treated by HT (HT-treated; 33.5°C±0.5°C for 72h starting approximately 6h after delivery) and 6 were not (non-HT-treated). Both CBV and ScO2 were measured by TRS at 6, 24, 48, and 72h after birth. Magnetic resonance imaging (MRI) was performed 1-2weeks after birth to assess brain injury. RESULTS: Five neonates had adverse outcomes (3 HT-treated, 2 non-HT-treated). Of these, 1 died within 3days of birth and 4 had abnormal MRI findings, including basal ganglia, white matter, and/or thalamic lesions. The other 6 neonates had normal MRI findings (favorable outcome). At 6h after birth, CBV was significantly higher in neonates with adverse outcomes compared with those with a favorable outcome. At 24h after birth, ScO2 was significantly higher in neonates with adverse outcomes. Furthermore, we found that combined CBV at 24h after birth plus ScO2 had the best predictive ability for neurological outcome: sensitivity, specificity, positive predictive value, and negative predictive value were all 100%. CONCLUSION: Early postnatal CBV and ScO2 elevations were predictive of a poor outcome in HIE. Therefore, measuring combined CBV plus ScO2 at 24h after birth can allow more precise prediction of neurological outcome. Control of postnatal CBV and ScO2 is critical for effective HIE treatment.

Cerebral blood volume measurement using near-infrared time-resolved spectroscopy and histopathological evaluation after hypoxic-ischemic insult in newborn piglets.

The aim of this study was to assess the relationship between the cerebral blood volume (CBV) measured by near-infrared time-resolved spectroscopy (TRS) and pathological change of the brain in a hypoxic-ischemic (HI) piglet model. Twenty-one anesthetized newborn piglets, including three sham controls, were studied. An HI event was induced by low inspired oxygen. CBV was measured using TRS (Hamamatsu TRS-10). Data were collected before, during, and 6h after the insult. CBV was calculated as the change from the end of the insult. The piglets were allowed to recover from anesthesia for 6h after the insult. At the age of 5 days, the brains of the piglets were perfusion-fixed, and histologic evaluations of brain tissue were performed. The extent of histopathological damage was graded in 0.5-unit intervals on a 9-step scale. CBV increments were well correlated with histopathological scores, especially at 1 and 3h after resuscitation. Spearman's rank-correlation coefficients at 1, 3, and 6h after resuscitation in the gray matter were 0.9016, 0.9127, and 0.6907, respectively. We conclude that an increased CBV after HI insult indicates more marked histological brain damage. CBV measurement immediately after resuscitation provides a more precise prediction of the histological outcome.

Inhibitory effects of AG490 on H2O2-induced TRPM2-mediated Ca(2+) entry.

Transient receptor potential melastatin 2 (TRPM2) is an oxidative stress-sensitive Ca(2+)-permeable channel that controls Ca(2+) signalling. The activation of Janus kinase 2 (Jak2) by oxidative stress is implicated in the production of inflammatory mediators. We found that AG490, a Jak2 inhibitor, had an inhibitory effect on H2O2-induced TRPM2 activation. The purpose of this study was to examine the underlying mechanisms of the inhibitory effects of AG490. Activation of TRPM2 in TRPM2-expressing human embryonic kidney 293 (TRPM2/HEK) cells or the human monocytic cell line U937 was monitored by fluorescence-based Ca(2+) imaging and patch-clamp techniques. Treatment with AG490 almost completely blocked H2O2-induced increase in intracellular Ca(2+) in TRPM2/HEK and U937 cells. In the patch-clamp study, AG490 inhibited the H2O2-evoked inward current but not the ADP-ribose-induced inward current in TRPM2/HEK cells. In contrast, Jak inhibitor 1 (pyridone 6) and staurosporine, both of which inhibit Jak2, had no effect on H2O2-induced increase in intracellular Ca(2+). Moreover, AG490 decreased intracellular reactive oxygen species level, which was measured by using a hydroperoxide-sensitive fluorescent dye, on incubation with H2O2. In the cell-free assay system, AG490 scavenged hydroxyl radicals but not H2O2. These findings indicate that AG490 significantly reduces H2O2-induced TRPM2 activation, presumably by scavenging hydroxyl radicals rather than Jak2-dependent mechanisms. Although transient receptor potential ankyrin 1 (TRPA1) channel is also activated by H2O2, the H2O2-induced Ca(2+) entry through TRPA1 was only slightly delayed by AG490. This validates the potential use of AG490, as one of the materials for characterizing the role of TRPM2 channels in pathological models.

Clinical effect of alpha 1D/A adrenoceptor inhibitor naftopidil on benign prostatic hyperplasia: an international prostate symptom score and King's Health Questionnaire assessment.

OBJECTIVES: To examine the effect of alpha 1D/A adrenoceptor inhibitor naftopidil on health-related quality of life (QOL) in men with benign prostatic hyperplasia (BPH). METHODS: A total of 56 newly diagnosed patients with symptomatic BPH were prospectively enrolled and treated with 50 mg naftopidil daily for more than 12 weeks. All underwent pre-treatment documentation of lower urinary tract symptoms, QOL assessment using the international prostate symptom score (IPSS) and King's Health Questionnaire (KHQ), and uroflowmetry. A post-treatment assessment was performed at 12 weeks. RESULTS: IPSS scores as well as QOL index showed a significant improvement after naftopidil administration. Similarly, all seven domains except general health perceptions and social limitations in the KHQ questionnaire were significantly improved. When dividing the patients into overactive bladder (OAB) and non-OAB groups, only the OAB group showed significant improvement in almost all the domains of KHQ. Change ratios of the IPSS were not associated with those of KHQ domain scores in the OAB group. On the other hand, in the non-OAB group more domains presented improvements, which were associated with those of IPSS scores. CONCLUSIONS: Twelve-week treatment with naftopidil for symptomatic BPH patients is associated with significant improvement in the IPSS, QOL index, maximum urinary flow rate, post-void residual urine volume (PVR) and almost all domains in KHQ. KHQ is useful for the evaluation of clinical response in BPH patients, particularly in those with associated OAB.

[The effectiveness of naftopidil in patients with benign prostatic hyperplasia evaluated by QOL index].

We examined the effectiveness of naftopidil in 81 patients with lower urinary tract symptoms (LUTS) caused by benign prostatic hyperplasia (BPH). We examined quality of life (QOL) and determined which symptoms improved as a result of naftopidil administration. The findings indicated that storage symptoms, voiding symptoms, total International Prostate Sympotom Score (IPSS), QOL index, Qmax and residual urine volume were significantly improved after treatment when compared to baseline. Improvement of nocturia and incomplete emptying by naftopidil contributed to improvement in QOL, odds ratio between the good response group and poor response group were 3.6 and 2.3, respectively. During naftopidil treatment, two of the 81 patients complained of adverse events. The results show that naftpidil is effective for LUTS caused by BPH, and that improvement of nocturia and incomplete emptying contributed to the improvement in QOL.

[Ileovesical fistula caused by bladder cancer with bladder stone].

Enterovesical fistula is a very rare complication of primary urological malignancies. A case of ileovesical fistula caused by a bladder carcinoma is presented. A 66-year-old male was referred with complaints of urinary pain. On admission, fecaluria and urinary tract infection with bladder stone were detected. Cystography revealed the passage of contrast medium into the small bowel. Under the diagnosis ofileovesical fistula due to suspected inflammatory disease, sigmoidectomy and segmental small bowel resection with partial cystectomy were performed. Histological evaluation revealed a poorly differentiated urothelial carcinoma. Without further treatment, the patient died from cancer five months after operation. However, it is hard to assess the effect of fistulas on prognosis. Since it has been reported that about 40% of the patients with T4 bladder tumors could be potentially cured with radical resection, we recommend a thorough examination to confirm the diagnosis of primary disease to obtain the best results.

Usefulness of tonsil forceps in radical retropubic prostatectomy.

The presence of positive surgical margins after radical retropubic prostatectomy (RRP) for prostate cancer leads to an increased risk of progression and reduces disease free survival. A positive surgical margin at the apex is more frequent and is associated with worse clinical prognosis compared to other locations. The urethra usually enters the prostate slightly anterior and proximal to the prostatic apex. After dividing the dorsal vessels and separating neurovascular bundles (NVB) from the prostatic urethral junction using scissors, the operator dissects around the urethra just below the apex to avoid incision into the apex and injury of the NVB and sphincter mechanism. We use tonsil forceps instead of a right-angle clamp to make this important operative step more approachable. Its special curved shape with an angle of 105 degrees and short tip should make it much easier to isolate the urethra just below the apex from the surrounding tissue.

Kidney stone disease and risk factors for coronary heart disease.

BACKGROUND: We conducted a case-control study to examine the impact of coronal heart disease (CHD) risk factors on calcium oxalate (CaOX) stone formation. METHODS: Variables included body mass index (BMI), current alcohol use, smoking habit, hypertension, hypercholesterolemia, diabetes mellitus, and hyperuricemia. Data suf fi cient for analysis were obtained for 181 CaOX stone formers and 187 controls. RESULTS: Seven of 181 stone formers (3.9%) had a history of CHD compared with none of 187 control subjects (P = 0.007). In univariate logistic regression analysis, smoking habit (OR 4.41, 95% CI 2.85-6.84, P < 0.0001), hypertension (OR 4.24, 95% CI 2.61-6.91, P < 0.0001), hypercholesterolemia (OR 3.03, 95% CI 1.77-5.20, P < 0.0001) and BMI (OR 1.10, 95% CI 1.04-1.17, P = 0.007) reached statistical signi fi cance. In a multivariate logistic regression analysis, smoking habit (OR 4.29, 95% CI 2.68-6.86, P < 0.0001), hypertension (OR 3.57, 95% CI 2.11-6.07, P < 0.0001), and hypercholesterolemia (OR 2.74, 95% CI 1.51-5.00, P = 0.001) reached statistical signi fi cance, while BMI (OR 1.06, 95% CI 0.99-1.12, P = 0.09) did not. CONCLUSIONS: CaOX stone formers are signi fi cantly associated with several CHD risk factors, including smoking habit, hypertension, hypercholesterolemia, and obesity.

Transplacental effects of allopurinol on suppression of oxygen free radical production in chronically instrumented fetal lamb brains during intermittent umbilical cord occlusion.

OBJECTIVE: To evaluate the transplacental effect of allopurinol, which acts as a xanthine oxidase inhibitor and free radical scavenger, on inhibiting the production of superoxides during intermittent partial umbilical cord occlusion. METHODS: Using four chronically instrumented fetal lambs, ewes received 400 mg allopurinol over a period of two hours. Concentrations of allopurinol and oxypurinol in blood samples from mothers and fetuses and fetal brain microdialysis perfusate were measured by HPLC. In another three cases the production of superoxide during intermittent umbilical cord occlusion was studied by measurement of chemiluminescence in perfusate before and after administration of Allopurinol. RESULTS: (i) Allopurinol concentration in mothers had reached equilibrium by 30 min after starting administration and maintained a concentration about 6 mug/ml. Allopurinol concentration in fetuses increased gradually and reached 2.25 +/- 0.54 microg/ml at 120 min; (ii) Oxypurinol concentration in both mothers and fetuses increased during administration of allopurinol; (iii) Concentrations of allopurinol and oxypurinol in the perfusates reached 0.32 +/- 0.12 microg/ml, 0.53 +/- 0.22 microg/ml at 120 min respectively; and (iv) Administration of allopurinol significantly suppressed superoxide production during intermittent partial umbilical cord occlusion. CONCLUSION: These results demonstrated a good transfer of allopurinol from mother to fetus and suggested the possibility of intrauterine treatment to inhibit fetal brain damage resulting from increased oxygen free radicals.

Do BPH patients have neurogenic detrusor dysfunction? A uro-neurological assessment.

INTRODUCTION: Most men with benign prostatic hyperplasia (BPH) are middle- aged and elderly. Neurogenic detrusor dysfunction (NDD) may also occur in these populations. We made an uro-neurological assessment in such patients in order to investigate the possibility of a concurrent NDD in patients diagnosed with BPH, and to correlate the presence of NDD with treatment outcome. MATERIALS AND METHODS: 28 men, mean age 66.4 years, referred by urologists at the outpatient clinic as BPH (initial diagnosis: BPH alone, 24, BPH and NDD, 4) with regards to digital examination and lower urinary tract symptoms, underwent urodynamic study and neurological examination. MRI scans were performed to confirm the diagnosis. RESULTS: Urodynamic study (except for 2, acontractile detrusor (AD)) showed equivocal obstruction (EO), 6; underactive detrusor (UD), 9 (voiding phase); detrusor overactivity (DO), 12; urethral relaxation, 1, and reduced sensation, 5 (filling phase). EO correlated with the presence of UD (p = 0.03). DO did not correlate with the prostate size or urodynamically-defined outlet obstruction. Reduced sensation was common in patients >65 years of age (p = 0.05). Neurological examination/imaging showed exaggerated reflexes, 1; decreased reflexes, 6; multiple cerebral infarction, 8; cervical spondylosis, 1 (in patients with DO); lumbar spondylosis, 5 (in patients with AD/UD), and the final diagnosis was made: definite BPH, 6; probable BPH, 9; BPH and NDD, 13, and NDD were common in the patients aged >65 years (p = 0.015). Transurethral prostatectomy and alpha-blocker were mostly successful, but the failure rate was reported in definite BPH, none, probable BPH, 29%, and BPH and NDD, 33%. CONCLUSIONS: BPH patients, particularly those >65 years of age, commonly have NDD. Multiple cerebral infarction (upper neuron disorder) and lumbar spondylosis (lower neuron disorder) might contribute to DO and UD, respectively. A uro-neurological assessment is important to select typical BPH patients for maximizing therapeutic benefit.

Pyogenic psoas abscess: difficulty in early diagnosis.

AIM: To report on the clinical features, diagnosis, and treatment of psoas abscess (PA) with special attention to the presence of septic shock. PATIENTS AND METHODS: This study included 17 patients (mean age 66.2, range 43-81 years) with PA. Treatment consisted of intravenous administration of antibiotics and abscess drainage, either surgical or percutaneous with ultrasound guidance. RESULTS: The typical patients presented with fever >38 degrees C (16/17, 94%), pain in back, flank, or abdomen (15/17, 88%), hip flexion contracture with pain extension (14/17, 82%), and mass felt in the flank (5/17, 29%). All 8 patients without septic shock (100%) had the clinical triad (fever, pain in back, flank, or abdomen, and hip flexion contracture) as compared with 4 of 9 patients with septic shock (44%) (p = 0.012). The duration of symptoms before hospitalization was significantly shorter in the patients with septic shock (median 2, range 1-5 days) than in those without septic shock (median 18.5, range 11-63 days; (p = 0.0005). The mortality rates were 33% (3 of 9) and 0% (0 of 8) in the patients with and without septic shock, respectively (p = 0.071). CONCLUSIONS: PA patients with septic shock had a tendency to have nonspecific symptoms and an occult clinical course as compared with those without septic shock. A delay in diagnosis and treatment can result in a worse clinical outcome (death or totally disabled state). Increased awareness of this condition should lead to earlier diagnosis and treatment with improved outcomes.