Inhibition of fibrotic changes in infrapatellar fat pad alleviates persistent pain and articular cartilage degeneration in monoiodoacetic acid-induced rat arthritis model.

OBJECTIVE: We have reported that fibrotic changes in infrapatellar fat pad (IFP) after acute joint inflammation are closely associated with persistent pain in rats. In this study, to examine the effects of anti-fibrotic treatment on persistent pain, we used C-type natriuretic peptides (CNP) at the recovery phase after acute joint inflammation. DESIGN: Thirty-two male Wistar rats were used in this study. Monoiodoacetic acid (MIA) was injected intra-articularly to induce IFP fibrosis and persistent pain. CNP was injected after acute inflammatory phase in the same knee joint. Time-course pain-avoidance behavior tests and histological analyses were performed to examine the effects of CNP. RESULTS: Histological evaluations indicated that intra-articular injection of CNP inhibited fibrotic changes in IFP after acute inflammation. Incapacitance tests indicated that MIA injection into rat knee joint quickly decreased the percent weight on ipsilateral limb. In the vehicle group, the decrease was maintained up to day 28, suggesting that pain persistence occurred after acute inflammation (Day 0/Day 28, Est Dif -8.15, CI -10.78∼-5.53, Linear mixed-effect model). In contrast, the pain was alleviated in the CNP group after day 14 (Day0/Day 14, -0.51, -2.62-1.59). In addition, we observed significant improvement in the degree of articular cartilage degeneration at day 14 in the CNP group (OARSI score: vehicle 16.14 ± 4.37 vs CNP 6.87 ± 3.44, P < 0.01; Wilcoxon rank sum test). CONCLUSION: Fibrotic changes in IFP may play important roles in both persistent pain and articular cartilage degeneration.

Management of paediatric tuberculosis in provincial and district hospitals in Afghanistan.

Case detection, diagnosis and treatment of tuberculosis 1 B) in children are challenging issues vorldwide. This study in Afghanistan aimed to evaluate paediatric TB case management, including contact investigation, at health facilities where all diagnostic processes were available. In 7 out of 8 regions of the country 1 province was selected. Documents used for management of paediatric TB cases were reviewed in 15 distinct hospitals and 8 provincial hospitals in the selected provinces. The key issues which emerged were: a low suspect rate among total outpatients (0.4%) and a very low suspect rate among children aged < 5 years; low performance of suspect management (68.5% suspects received further examinations); low utilization of other diagnostic methods; a high early defaulter rate (14.0%); and insufficient coverage of contact management (74.0%). This survey indicated that the Afghanistan national TB programme needs to develop plans to improve the quality of diagnosis, suspect management and contact management in paediatric TB cases.

Inferior Healing Rate in Isolated Meniscal Repair than that in Meniscal Repair with Concomitant ACL Reconstruction Evaluated with MRI.

Introduction: Isolated meniscal repair has been suggested as one of the contributing factors in unhealed meniscal repair. The purpose of this study was to compare the healing rate between isolated meniscal repair and meniscal repair with concomitant anterior cruciate ligament reconstruction (ACLR) using a standardised assessment method after propensity score matching. Materials and methods: Accuracy of the Crues' grading system for meniscal healing was validated using second-look arthroscopy as the reference standard in 17 patients. Propensity score matching (one-to-one) was performed between 26 patients who underwent isolated meniscal repair and 98 patients who underwent meniscal repair with concomitant ACLR. Patients were matched for sex, age, side and zone of the meniscal repair, and number of sutures. Healing rates at one year which were evaluated with magnetic resonance imaging (MRI) were compared between the two groups. Results: The sensitivity and specificity of the Crues' grading system on multiple plane MRI for meniscal healing were 100% and 83.3%, respectively. Both the isolated meniscal repair group and the meniscal repair with concomitant ACLR group included 21 patients after propensity score matching. Baseline characteristics did not differ significantly between the two groups. The healing rate was significantly lower in the isolated meniscal repairs group (14.3%) than in the meniscal repair concomitant with ACLR group (47.6%, P=0.04). Conclusion: The healing rate for isolated meniscal repair using a standardised MRI assessment method was inferior to that of meniscal repair with concomitant ACLR after propensity score matching.

Association of genetic variants for susceptibility to obesity with type 2 diabetes in Japanese individuals.

AIMS/HYPOTHESIS: In populations of East Asian descent, we performed a replication study of loci previously identified in populations of European descent as being associated with obesity measures such as BMI and type 2 diabetes. METHODS: We genotyped 14 single nucleotide polymorphisms (SNPs) from 13 candidate loci that had previously been identified by genome-wide association meta-analyses for obesity measures in Europeans. Genotyping was done in 18,264 participants from two general Japanese populations. For SNPs showing an obesity association in Japanese individuals, we further examined diabetes associations in up to 6,781 cases and 7,307 controls from a subset of the original, as well as from additional populations. RESULTS: Significant obesity associations (p < 0.1 two-tailed, concordant direction with previous reports) were replicated for 11 SNPs from the following ten loci in Japanese participants: SEC16B, TMEM18, GNPDA2, BDNF, MTCH2, BCDIN3D-FAIM2, SH2B1-ATP2A1, FTO, MC4R and KCTD15. The strongest effect was observed at TMEM18 rs4854344 (p = 7.1 × 10(-7) for BMI). Among the 11 SNPs showing significant obesity association, six were also associated with diabetes (OR 1.05-1.17; p = 0.04-2.4 × 10(-7)) after adjustment for BMI in the Japanese. When meta-analysed with data from the previous reports, the BMI-adjusted diabetes association was found to be highly significant for the FTO locus in East Asians (OR 1.13; 95% CI 1.09-1.18; p = 7.8 × 10(-10)) with substantial inter-ethnic heterogeneity (p = 0.003). CONCLUSIONS/INTERPRETATION: We confirmed that ten candidate loci are associated with obesity measures in the general Japanese populations. Six (of ten) loci exert diabetogenic effects in the Japanese, although relatively modest in size, and independently of increased adiposity.

Mitogen-activated protein kinase cascade is activated in glomeruli of diabetic rats and glomerular mesangial cells cultured under high glucose conditions.

The activation of protein kinase C (PKC) found in diabetic glomeruli and glomerular mesangial cells cultured under high glucose conditions has been proposed to contribute to the development of diabetic nephropathy. However, the abnormalities distal to PKC have not been fully elucidated yet. Herein, we provide the evidence that mitogen-activated protein kinase (MAPK) cascade, an important kinase cascade downstream to PKC and an activator of cytosolic phospholipase A2 (cPLA2) by direct phosphorylation, is activated in glomeruli isolated from streptozotocin-induced diabetic rats. MAPK cascade was also activated in glomerular mesangial cells cultured under high glucose (27.8 mmol/l) conditions for 5 days, and the activation of MAPK cascade was inhibited by treating the cells with calphostin C, an inhibitor of PKC. Furthermore, the activities of cPLA2 also increased in cells cultured under the same conditions and this activation was inhibited by both calphostin C and PD 098059, an inhibitor of MEK (MAPK or extracellular signal-regulated kinase [ERK] kinase). These results indicate that MAPK cascade is activated in glomeruli and mesangial cells under the diabetic state possibly through the activation of PKC. Activated MAPK, in turn, may induce various functional changes of mesangial cells at least through the activation of cPLA2 and contribute to the development of diabetic nephropathy.

Hepatoprotective action of adenovirus-transferred HNF-3gamma gene in acute liver injury caused by CCl(4).

Hepatocyte nuclear factor-3gamma (HNF-3gamma) is an important regulator of liver-specific genes and the expression of this factor is reduced in the liver injured by carbon tetrachloride (CCl(4)). Wistar rats were infected with a recombinant adenovirus carrying the cDNA for HNF-3gamma (AxCAHNF3gamma) via the tail vein and were treated with CCl(4) by intraperitoneal injection. Liver damage, such as swelling of the hepatocytes and increases in serum marker enzymes were markedly alleviated by AxCAHNF3gamma infection. Interestingly, hepatocyte growth factor (HGF) was strongly induced in the AxCAHNF3gamma-infected liver. Likewise, HNF-1alpha and HNF-1beta levels were increased, but HNF-3alpha and HNF-3beta levels were depressed in the liver. Our results suggest that the transduced HNF-3gamma gene leads to a hepatoprotective effect via the induction of HGF by the combined actions of liver-enriched transcription factors.

In vivo transfer of hepatocyte growth factor gene accelerates proliferation of hepatic oval cells in a 2-acetylaminofluorene/partial hepatectomy model in rats.

To clarify the effect of hepatocyte growth factor (HGF) on proliferation of hepatic oval cells, we transferred HGF gene into liver of the Solt-Farber rat model. Male Fisher 344 rats were infected with a recombinant adenovirus carrying the cDNA for HGF (pAxCAHGF) from tail vein. HGF mRNA showed its peak at 4 days, and diminished thereafter. The total and proliferating cell nuclear antigen-positive hepatic oval cells were significantly elevated in HGF-transferred rats, in which stem cell factor and c-kit mRNA increased at each time point. Our results suggest that in vivo transfer of the HGF gene into liver accelerates proliferation of hepatic oval cells in the Solt-Farber model in rats.

Intracerebral transplantation of the A7 immortalized astrocytic cell line.

The A7 cell line is an astrocyte-like cell immortalized by SV40 large T antigen, using retroviral-mediated gene transfer. These cells were transplanted into rat brains, and the graft-host interaction was investigated immunohistochemically. The A7 cells survived focally 2, 6 and 8 weeks after transplantation and retained the immunocytochemical properties observed in vitro. No immunological response was observed. GAP-43 and N-cadherin immunoreactivities were not expressed by A7 cells, but were seen in the matrix within the area of the graft and in the surrounding brain tissue. This indicates that A7 cells may stimulate expression of GAP-43 and N-cadherin immunoreactivity by host tissue. Expression of Thy 1.1 was not observed within the graft site after 2 weeks of survival, but 6 and 8 weeks after transplantation Thy 1.1 was observed within the graft area, indicating the possible co-existence of grafted cells and host tissue. Although indirect, these observations suggest that the A7 cells induce changes in host brain, including possible growth or regeneration of host tissue into the graft area.

Immunological reactions following intracerebroventricular transplantation of adrenal medulla.

Immunological reactions after intracerebroventricular syn-, allo- and xenogenic transplantation of adrenal medulla were investigated histologically. In xenografts only, T cell infiltration and graft rejection were observed. Syngrafts and allografts were not rejected and were not infiltrated by T cells, although expression of MHC class II antigen was observed at all survival times. Major histocompatibility complex (MHC) class I immunoreactivity was strongly expressed in adrenal cortex syngrafts, which could play a role in the rejection of grafts containing mixed cell populations. The survival of chromaffin cells in allografts was decreased as compared to syngrafts, and there were fewer allograft animals with large numbers of surviving chromaffin cells. There was some increased cellularity (microglia and macrophages) in allografts even though no T cell infiltration was found. Therefore, it appears that this limited survival of intracerebral adrenal medulla allografts is not due to T cell-mediated graft rejection.

Human cortical neuronal cell line (HCN-1): further in vitro characterization and suitability for brain transplantation.

The human neuronal cell-1 (HCN-1) line has recently been established. Under favorable conditions, these cells differentiate into mature neuronal phenotypes. Here we report on further characterization of these cells. Cultured HCN-1 cells express fibronectin immunoreactivity and grow well on fibronectin substrate but do not respond to human bFGF. In the undifferentiated state, some HCN-1 cells show MHC class I antigen expression. After differentiation, HCN-1 cells and their processes are MHC class I negative. On the other hand, interferon-gamma stimulation enhances MHC class I expression but does not induce MHC class II immunoreactivity. Our in vitro data indicate that HCN-1 cells express mixed characteristics, including both neuronal and mesenchymal markers, and are consistent with the suggestion that the HCN-1 cell line resembles an immature neuroepithelial cell precursor with a complex origin. One possible application of the use of the HCN-1 cells includes intracerebral transplantation. We also examined the survival of dissociated HCN-1 cells implanted into rat brain parenchyma. The host animals were not immunosuppressed. Despite expression of MHC class I antigens, small clusters of HCN-1 cells survived in the rat brain. These xenografts did not induce distinct immunological responses within the host brain tissue. Surviving HCN-1 cells demonstrated similar features to those observed in culture. Our preliminary results suggest that the HCN-1 cell line would be suitable for intracerebral transplantation in primates or humans. However, it may be that short-term host immunosuppression or addition of HCN-1 cell differentiation factors would be beneficial for enhanced cell survival.

Mechanism of the progression of diabetic nephropathy to renal failure.

Although the evolution of diabetic nephropathy is brought about mostly by persistent hyperglycemia, its progression may be influenced by various other factors such as hypertension and dietary protein intake. It has been recently suggested in the literature that the gene polymorphism of angiotensin converting enzyme (ACE) might be associated with the development of diabetic nephropathy, because the DD genotype of ACE gene is closely associated with the presence of nephropathy in diabetic subjects. However, in our present analysis the frequency of the DD genotype in patients with non-insulin dependent diabetes is not significantly related to the presence or absence of nephropathy. It remains to be clarified by multi-center analysis using large numbers of patients whether the gene polymorphism of ACE is related to the progression of diabetic nephropathy to renal failure. Furthermore, it has been postulated that the interstitial fibrosis evaluated in renal biopsy specimens is significantly correlated with the declining of renal function in diabetic patients. However, it is not possible to clinically quantitate the interstitial fibrosis without performing renal biopsy. We have recently found that the urinary excretion of type IV collagen is significantly increased in diabetic patients. Moreover, the increase in urinary type IV collagen is well correlated with the amount of urinary albumin. Since type IV collagen in the urine is probably derived from tubulointerstitial tissue, it is likely that the increased amount of type IV collagen in the urine may reflect the fibrotic change in diabetic kidneys. Whether the increase in urinary type IV collagen is able to predict for the progression of diabetic nephropathy in the future should be examined.

Evidence of apoptotic procedure in deafferented striatum after cortical injury in young adult rats.

In denervated striatum after excitotoxic cortical lesion in young adult rats, apoptotic cells, though quite few, were observed by TUNEL 2 weeks after surgery. Also, prominent expressions of p53 were observed at the same time. These data indicate that apoptotic procedure may be involved in the denervation-induced degeneration even in young adults.

The exogenous control of transfected c-fos gene expression and angiogenesis in cells implanted into the rat brain.

Previously, we established a stable transfectant, Nf-1, from normal rat kidney (NRK) fibroblasts transfected with a human metallothionein II A (hMT-IIA) promoter/human genomic c-fos fusion gene to produce c-Fos protein. Since the hMT-IIA promoter can be activated by heavy metals, the level of human c-fos gene expression can be increased by addition of heavy metals to the culture medium of Nf-1 cells and the anchorage-independent growth of Nf-1 in soft agar is markedly enhanced in the presence of transforming growth factor-beta (TGF-beta) and epidermal growth factor (EGF). In this study, we found that the hMT-IIA promoter can be activated by zinc, resulting in the elevation of fused c-fos gene expression in Nf-1 cells. We transplanted NRK and Nf-1 cells into the striatum of the rat brain and investigated whether expression of the human c-fos gene could be modified in the brain by exogenous zinc. After 8 weeks, we found that the Nf-1 cells could survive in the rat brain without any immunosuppression and grafts of Nf-1 induced angiogenesis when zinc was administered. Such implants enhanced the expression of c-fos mRNA by zinc. These results indicated that the transplanted cells continued expressing the c-fos transgene when the rats were given drinking water containing zinc, resulting in the promotion of cell growth and of neovascularization. This study will present a useful animal model of gene therapy by control of transgene expression in the brain.

Beraprost sodium, an analogue of prostacyclin, induces the expression of mitogen-activated protein kinase phosphatase and inhibits the proliferation of cultured mesangial cells.

Beraprost sodium, an analogue of prostacyclin, increases intracellular cyclic adenosine monophosphate (cAMP) in cultured glomerular mesangial cells. We examined the effect of beraprost on mesangial cell proliferation. Beraprost was able to inhibit fetal bovine serum-stimulated proliferation of mesangial cells in concentrations enough to increase cellular cAMP. By northern blot analysis, beraprost induced the expression of MKP-1, a mitogen-activated protein kinase phosphatase, in a dose- and time-dependent manner, similarly to dibutyryl cAMP and adrenomedullin. These results indicate that beraprost inhibits the proliferation of mesangial cells and one of the mechanisms might be cAMP-dependent induction of MKP-1.

Microalbuminuria is not associated with cardiovascular death in Japanese NIDDM.

To evaluate whether the presence of microalbuminuria can predict cardiovascular death in Japanese subjects with non-insulin-dependent diabetes mellitus (NIDDM), we investigated 297 Japanese NIDDM patients with Albustix-negative urine. Patients were divided into two groups, normoalbuminuric (n = 201) and microalbuminuric (n = 96) and followed until death or the end of 1994 (the mean follow-up period was 6.4 years). During the follow-up period, 28 deaths (14 normoalbuminuric and 14 microalbuminuric patients) were confirmed and only 10 deaths were attributed to cardiovascular disease (6 normoalbuminuric and 4 microalbuminuric patients). Although the age- and sex-adjusted mortality rate from all-causes in the microalbuminuric group was significantly higher than that in the normoalbuminuric group (13.5 vs. 8.2 per 1000 person-years: P < 0.05), the mortality rate from cardiovascular disease was not significantly different between two groups (3.4 vs. 3.3 per 1000 person-years). On age-adjusted Cox proportional hazards analysis. HbA1c and triglyceride were independent risk factors in mortality from cardiovascular disease, while microalbuminuria was not associated with cardiovascular death. These results indicate that, unlike Caucasians, the presence of microalbuminuria can not predict cardiovascular death in Japanese subjects with NIDDM.

Neurocutaneous vascular hamartomas mimicking Cobb syndrome. Case report.

The authors report the rare case of a patient with neurocutaneous vascular hamartomas mimicking Cobb syndrome. An 8-year-old boy was admitted to the authors' hospital with progressive urinary disturbance and upper back pain. Multiple skin nevi had been noted at the child's birth. Radiological examination revealed multiple cavernous angiomas in the spinal cord in the same metamere in which the skin nevi had been observed and also in the left cerebral hemisphere. His symptoms gradually improved without surgical intervention. Four years later he was readmitted because of a cerebral hemorrhage involving the left cerebral peduncle. Nonsurgical treatment was chosen because his symptoms promptly improved. To the best of the authors' knowledge, this is the first case of multiple cavernous angiomas in the brain and spinal cord associated with skin nevi. The authors discuss this clinical entity and the significance of the disease.

Observation of the external aperture of the vestibular aqueduct using three-dimensional surface reconstruction imaging.

Observation of the external aperture of the vestibular aqueduct was hitherto possible only in cadavers or dry temporal bones; however, by applying three-dimensional surface reconstruction imaging, it is now possible to observe solid-looking images of this structure in living humans. When the width of the external aperture of the vestibular aqueduct was measured in 58 people, it was found to be significantly narrower in the affected ears of patients with Meniere's disease than in normal ears.

Clinical features of symptomatic Rathke's cleft cyst.

To investigate the clinical features of Rathke's cleft cysts (RCCs), we retrospectively analyzed 15 cases with histologically confirmed RCCs. All patients underwent formal testing of visual field, endocrinological evaluation and magnetic resonance imagings. As overall presenting symptoms, endocrine disturbance was the most common symptoms, followed by visual disturbance and headache. Among the endocrine disturbances based on adenohypophysial dysfunction, hyperprolactinemia was most common. Considering the size of RCCs, RCCs could induce hyperprolactinemia only when the cysts became large enough to compress the infundibular system. Our series showed relative high incidence of pituitary dwarfism and diabetes insipidus (DI). These facts indicated that RCCs could evoke hyposecretion of growth hormone in young patients and DI in aged patients by direct compression of the pituitary gland in the early stage of progression. All cases who had headache had no other symptoms. We could not prove the evidence that RCCs could induce headaches in these cases. This might be suggested that headache could not be a sole symptom in cases of RCCs.

An objective evaluation method for facial mimic motion.

This technique was designed to establish a simple, objective evaluation system for facial paralysis through the use of a personal computer. A total of 24 marks were placed on the faces of subjects for the following procedures. Movements of the face were photographed with a video-camera and fed continuously into the computer. Ten frames per movement representing facial movement from rest to maximum movement were selected for analysis. By means of a digital image-processing technique, only the marks placed on the face were extracted, and the movement of these marks was quantitatively analyzed. A total of 44 healthy subjects with no history of facial paralysis were used as a normal control group. The patients with facial paralysis consisted of nine subjects with Bell's palsy and three with Ramsay Hunt syndrome. In the eye-closing motions, no significant differences were found between the sum of the movement distances on the left and right sides in each normal subject. However, the patients with facial paralysis showed distinct differences from those obtained in the normal subjects. The improvement process was also evaluated with a ratio of affected- and normal-side facial movements.

Midaortic syndrome in childhood associated with a ruptured cerebral aneurysm: a case report.

BACKGROUND: We present a patient with a midaortic syndrome who presented with subarachnoid hemorrhage caused by rupture of an anterior communicating artery aneurysm. CASE DESCRIPTION: A 14-year-old boy with midaortic syndrome was admitted to our hospital because of subarachnoid hemorrhage due to rupture of an anterior communicating artery aneurysm. He also developed acute renal failure due to previously controlled hypotension. After blood dialysis, successful clipping of the aneurysm was performed. The postoperative course was complicated by malignant renovascular hypertension due to midaortic syndrome. Medical treatment failed to control his hypertension; left primary nephrectomy improved his condition. CONCLUSION: Although midaortic syndrome is rare, it may be significant as a cause of cerebral hemorrhage in childhood.