Sex-based differences in CD103+ dendritic cells promote female-predominant Th2 cytokine production during allergic asthma.

BACKGROUND: Gender disparities in adult patients with asthma regarding its prevalence and severity are mainly due to enhanced type 2 T-helper (Th2) cytokine production in female patients compared to that in male patients. However, the pathways mediating this effect remain unclear. OBJECTIVE: We aimed to determine the roles of two major subsets of dendritic cells (DCs) in females, specifically those displaying CD11b or CD103, during enhanced Th2 priming after allergen exposure, using an ovalbumin-induced asthma mouse model. METHODS: Sex-based differences in the number of DCs at inflamed sites, costimulatory molecule expression on DCs, and the ability of DCs to differentiate naïve CD4+ T cells into Th2 population were evaluated after allergen exposure in asthmatic mice. In addition, we assessed the role of 17ß-oestradiol in CD103+ DC function during Th2 priming in vitro. RESULTS: The number of CD11bhigh DCs and CD103+ DCs in the lung and bronchial lymph node (BLN) was increased to a greater extent in female mice than in male mice at 16 to 20 hours after ovalbumin (OVA) inhalation. In BLNs, CD86 and I-A/I-E expression levels and antigen uptake ability in CD103+ DCs, but not in CD11bhigh DCs, were greater in female mice than in male mice. Furthermore, CD4+ T cells cultured with CD103+ DCs from female mice produced higher levels of interleukin (IL)-4, IL-5, and IL-13, compared with CD4+ T cells cultured with CD103+ DCs from male mice. The 17ß-oestradiol-oriented enhancement of CD86 expression on CD103+ DCs after allergen exposure induced the enhanced IL-5 production from CD4+ T cells. CONCLUSIONS AND CLINICAL RELEVANCE: These findings suggest that with regard to asthma, enhanced Th2 cytokine production in females might be attributed to 17ß-oestradiol-mediated Th2-oriented CD103+ DCs in the BLN.

Distribution of natriuretic peptide receptor-C immunoreactivity in the rat brainstem and its relationship to cholinergic and catecholaminergic neurons.

The natriuretic peptide receptor type C (NPR-C) binds all natriuretic peptides. It is thought to be involved in the clearance of natriuretic peptides and more recently has been defined as essential for the neuromodulatory effects of natriuretic peptides. Although the distribution of NPR-C mRNA has been reported in the rat forebrain, there are no data on the distribution of NPR-C in the brainstem. We report an immunofluorescence study on the distribution of NPR-C immunoreactivity in the rat brainstem, and its presence in cholinergic and catecholaminergic neurons. NPR-C immunoreactivity was detected in several regions, including the periaqueductal gray, oculomotor nucleus, red nucleus and trochlear nucleus of the midbrain; the pontine nucleus, dorsal tegmental nucleus, vestibular nucleus, locus coeruleus, trigeminal motor nucleus, nucleus of the trapezoid body, abducens nucleus and facial nucleus of the pons; and the dorsal motor nucleus of the vagus, hypoglossal nucleus, lateral reticular nucleus, nucleus ambiguus and inferior olivary nucleus of the medulla oblongata. Interestingly, NPR-C immunoreactivity was detected in the cholinergic neurons of the oculomotor nucleus, trochlear nucleus, dorsal tegmental nucleus, motor trigeminal nucleus, facial nucleus, dorsal motor nucleus of the vagus, nucleus ambiguus and hypoglossal nucleus. Furthermore, NPR-C immunoreactivity was detected in several catecholaminergic neuronal groups including the A6, A5, A1, C3 and C1 cell groups. These results are consistent with an important role for natriuretic peptides in neuroendocrine regulation and central cardiovascular integration. The extensive distribution of NPR-C in the brainstem supports the hypothesis that NPR-C is involved in the neuromodulatory effect of natriuretic peptides.

Specific p53 gene mutations in urinary bladder epithelium after the Chernobyl accident.

After the Chernobyl accident, the incidence of urinary bladder cancers in the Ukraine population increased gradually from 26.2 to 36.1 per 100,000 between 1986 and 1996. Urinary bladder epithelium biopsied from 45 male patients with benign prostatic hyperplasia living in radiocontaminated areas of Ukraine demonstrated frequent severe urothelial dysplasia, carcinoma in situ, and a single invasive transitional cell carcinoma, combined with irradiation cystitis in 42 cases (93%). No neoplastic changes (carcinoma in situ or transitional cell carcinoma) were found in 10 patients from clean areas (areas without radiocontamination). DNA was extracted from the altered urothelium of selected paraffin-embedded specimens that showed obviously abnormal histology (3 cases) or intense p53 immunoreactivity (15 cases), and mutational analysis of exons 5-8 of the p53 gene was performed by PCR-single-strand conformational polymorphism analysis followed by DNA sequencing. Nine of 17 patients (53%) had one or more mutations in the altered urothelium. Urine sediment samples were also collected from the patients at 4-27 months after biopsy and analyzed by PCR-single-strand conformational polymorphism analysis or yeast functional assay, and identical or additional p53 mutations were found in four of five cases. Interestingly, a relative hot spot at codon 245 was found in five of nine (56%) cases with mutations, and 11 of the 13 mutations determined (73%) were G:C to A:T transitions at CpG dinucleotides, reported to be relatively infrequent (approximately 18%) in human urinary bladder cancers. Therefore, the frequent and specific p53 mutations found in these male patients may alert us to a future elevated occurrence of urinary bladder cancers in the radiocontaminated areas.

An interleukin-1 binding region oligopeptide from the human type I IL-1 receptor reduces acute inflammation, delayed hypersensitivity reaction, and lethal endotoxemia in animals.

The effect of an interleukin-1 binding region oligopeptide from the interleukin-1 receptor on various inflammatory responses was investigated in animal models. A synthetic peptide (KICIRIQIS) corresponding to 86-93 of the extracellular domain of the human type I interleukin-1 receptor was used. Carrageenan-induced rat paw edema, a model of acute inflammation, was dose dependently suppressed by intraperitoneal administration of the peptide. The delayed hypersensitivity reaction to sheep red cells was diminished by pretreatment of mice with the peptide at a relatively high dose. In a murine lethal endotoxemia model, animals treated with the interleukin-1 receptor peptide (10 mg/kg x 4) showed significantly better survival than vehicle-treated animals when the peptide was administered from 20 minutes after lipopolysaccharide injection. Improved survival was accompanied by suppression of lipopolysaccharide-induced production of colony-stimulating factor, although the peptide did not improve hypoglycemia. These findings suggest that the interleukin-1 receptor peptide may be a potential treatment for various inflammatory processes.

Further evidence that the mouse defense test battery is useful for screening anxiolytic and panicolytic drugs: effects of acute and chronic treatment with alprazolam.

The Mouse Defense Test Battery (MDTB) has been designed to investigate defensive responses of Swiss-Webster mice confronted with a natural predator, a rat. These behaviors include flight, avoidance, defensive threat/attack responses, and risk assessment activities. Previous studies with the MDTB have suggested that this model may have some utility for the investigation of panicogenic and antipanic compounds. In the present study the MDTB was used to investigate the effects of acute (0.05-1 mg/kg, i.p., 30 min) or chronic (0.5-2 mg/kg, one daily i.p. injection during 10 days) treatment with the benzodiazepine receptor (BZPR) full agonist and panicolytic agent alprazolam. At non motor-impairing doses (0.05-0.5 mg/kg), acute alprazolam failed to alter the avoidance distance between the subject and the predator, the number of avoidances when the rat is approaching, predator assessment activities, defensive threat/attack responses when contact is forced between the subject and the predator or contextual escape attempts after the predator was removed. This was in contrast to chronic treatment which decreased both avoidance variables at 0.5 and 1 mg/kg, defensive threat/attack responses at all doses, and predator assessment responses at 0.5 mg/kg. In addition, the latter treatment reduced post-predator potentiation of escape attempts at 2 mg/kg. These results (1) confirm previous findings with the BZPR full agonist chlordiazepoxide, indicating that these compounds generally attenuate antipredator defensive responses in Swiss-Webster mice; (2) support recent data indicating that panic-altering drugs modulate flight/escape reactions, and suggest that the primary mechanism of action of drugs with efficacy against panic disorder may involve neural systems controlling flight; (3) confirm that the MDTB may be useful for the investigation of panicolytic as well as anxiolytic agents.

Dimethylarsinic acid induces 8-hydroxy-2'-deoxyguanosine formation in the kidney of NCI-Black-Reiter rats.

Dirnethylarsenic peroxyl radical [(CH(3))(2)AsOO] has been postulated to be responsible for DNA damage induced by dimethylarsinic acid (DMA). In an effort to elucidate the possible mechanism of tumor-inducing potential of DMA, an experiment was designed to investigate the formation of 8-hydroxy-2'-deoxyguanosine (8-OHdG), a specific marker of oxidative base damage in the kidney tissues of NCI-Black Reiter (NBR) rats. Animals were divided into four groups and administered the vehicle - saline, 5, 10 and 20 mg/kg body weight respectively of DMA by gavage, once a day, 5 days a week, for a period of 4 weeks. DMA induced increase of 8-OHdG levels in the kidney of the rats treated, with the highest level at the dose of 10 mg/kg body weight. Analysis of the kidney for cell proliferation employing PCNA-positive index showed greater proliferation in the tissues of treated rats. However, DMA did not have any influence on apoptosis in this regimen. Histopathological examination of the kidney selections revealed the presence of vacuolated degeneration and dilation of the proximal tubule cells in two groups (10 and 20 mg/kg body weight). This study provides evidence to substantiate the role of DMA in inducing oxidative DNA damage in the kidney.

Presence of a no-observed effect level for enhancing effects of development of the alpha-isomer of benzene hexachloride (alpha-BHC) on diethylnitrosamine-initiated hepatic foci in rats.

The dose dependence of the promoting effects of the alpha-isomer of benzene hexachloride (alpha-BHC) on hepatocarcinogenesis was investigated in a medium-term rat liver bioassay (Ito test). A total of 195 F344 male rats, 6 weeks old, were given a single intraperitoneal injection of diethylnitrosamine (DEN) at the start of the experiment and subjected to two-thirds partial hepatectomy at week 3. Two weeks after the administration of DEN, alpha-BHC were fed to rats at doses of 0, 0.01, 0.1, 0.5, 1, 2, 4, 7.5, 15, 30, 60, 125 and 500 ppm in diet for 6 weeks. All surviving animals were killed at week 8, and their livers were examined immunohistochemically for detection of glutathione S-transferase placental form (GST-P)-positive foci, surrogate preneoplastic lesions. Quantitative values for numbers and areas were dose-dependently increased in rats given alpha-BHC at 0.5-500 ppm. However, those for groups treated with 0.01 and 0.1 ppm were decreased, albeit not significantly in comparison to the controls. Cytochrome P450 3A2 (CYP3A2) protein levels and activities showed a good correlation to the number and area of GST-P-positive foci. These results support evidence of hormesis and indicate a no-observed effect level for alpha-BHC promoting potentials may exist regarding rat liver carcinogenesis, which correlates with expression of CYP3A2 in the liver.

5-HT1A agonists modulate mouse antipredator defensive behavior differently from the 5-HT2A antagonist pirenperone.

The mouse defense test battery (MDTB) has been designed to investigate defensive reactions in Swiss-Webster mice to situations associated with a natural predator, the rat, such as flight, avoidance, defensive threat, defensive attack, and risk assessment activities. The present study evaluated the ability of 8-OH-DPAT (0.05-10 mg/kg, SC, 5) and gepirone (2.5-10 mg/kg, IP, 30), a full- and a partial agonist at 5-HT1A sites, as well as pirenperone (0.25-1 mg/kg, IP, 30), a preferential 5-HT2A receptor antagonist, to exert an anxiolytic-like action in the MDTB. The most consistent effect of both 5-HT1A receptor agonists across tests was a marked reduction in predator assessment activity and defensive attack behavior. In contrast, neither of the two ligands was able to reduce flight responses to the approaching predator, and both failed to reduce in a specific manner contextual defense behaviors after the predator was removed. The 5-HT2A receptor antagonist pirenperone did not provide significant indication of an anxiolytic effect on predator assessment activity and postpredator potentiation of contextual defense responses, and had negligible influence on antipredator defensive behavior. The most interesting exception to this profile was a dose-related reduction in flight-related measures. In view of previous results indicating that the panic-promoting drug yohimbine increases flight/escape reactions and that the panicolytic compound alprazolam reduces these responses, we tentatively suggest that the preferential 5-HT2A receptor antagonist pirenperone may have some efficacy in improving panic attacks. In addition, the lack of effect of the 5-HT1A receptor agonists on these flight responses is consistent with clinical findings indicating that these agents are of limited use in the treatment of panic disorder. These findings suggest that the MDTB provides behavioural measures capable of differentiating between various classes of antianxiety drugs.

Non-carcinogenicity, but dose-related increase in preneoplastic hepatocellular lesions, in a two-year feeding study of phenobarbital sodium in male F344 rats.

Phenobarbital sodium (PB) was administered at dietary levels of 0 (control), 8, 30, 125 and 500 ppm to groups of 20 male F344/DuCrj rats for 104 weeks. There were no treatment-related clinical signs or adverse effects on survival rate, body weights, food consumption, and haematology or blood biochemistry data. Statistically significant increases of relative liver weights were found in the 500 and 125 ppm, but not the 30 and 8 ppm groups. Quantitative analysis of glutathione S-transferase placental form positive (GST-P+) hepatocyte foci/areas revealed clear increases limited to the 500 and 125 ppm groups. Western blotting revealed CYP2B1, 2C6 and 3A2 proteins to be also increased only with these high doses. In addition, significant increase of regenerative hepatocellular hyperplasias was noted in the 500 ppm group. No hepatocellular adenomas were observed, but a hepatocellular carcinoma arose in single rats of the 8 ppm and 125 ppm groups. No treatment-related changes were found in any other organs or tissues. Thus, under the experimental conditions used, the highest dose of PB (500 ppm) was not carcinogenic in male F344 rats. Furthermore, increase in putative preneoplastic proliferative hepatocytic lesions was only noted with 500 and 125 ppm.

Idiopathic sclerosing peritonitis in a man.

Idiopathic sclerosing peritonitis (ISP) is a rare condition which has been mainly reported in young adolescent women as a cause of small bowel obstruction. In these patients the small bowel is sometimes encased in a fibrous sac called an "abdominal cocoon". We describe a 62-year-old man who underwent exploratory laparotomy for ascites and abdominal mass. Laparotomy showed 5.4 l of ascites and the entire small bowel was encased in a fibrous sausage-like cocoon. The pathological findings were characteristic of sclerosing peritonitis.

An autopsy report of graft-versus-host disease (GVHD) following blood transfusion.

A case of graft-versus-host disease (GVHD) associated with blood transfusion in a 71-year-old man is reported. The patient received transfusions (irradiation red cells M.A.P 14 units, fresh frozen plasma 11 units) and developed features of GVHD including fever, a skin rash, diarrhea, liver dysfunction, hypoplastic bone marrow, and pancytopenia. He died on day 32 posttransfusion. Pathologically, lymphocyte infiltration of skin and liver lesions, degeneration of small bile ducts, crypt cell necrosis in the gastrointestinal tract, and bone marrow hypoplasia were observed. Immunohistochemically, infiltrating lymphocytes were CD8+ T type, the result suggesting that they role a part in posttransfusion GVHD.

[Experience with partial nephrectomy for renal cell carcinoma].

From Sept. 1991 to Jan. 1999, we performed partial nephrectomy on 7 patients with renal cell carcinoma. The indication was imperative for 3 patients, and elective for 4 patients. The 3 imperative cases consisted of bilateral renal cell carcinomas, a polycystic kidney disease and a contralateral atrophic kidney. All 4 patients with elective indication revealed renal cell carcinoma with a normal functioning contralateral kidney. The tumor size ranged from 1.3 cm to 6.0 cm (2.7 cm on average). The mean clamping time of renal artery was 22 minutes and mean blood loss was 400 ml. The pathological stage was pT1a in 6 patients and pT1b in one patient. Postoperative follow-up ranged from 4 months to 92 months (mean: 43 months). One patient with bilateral renal cell carcinoma died of metastases to the lungs and brain at 25 months postoperatively. The remaining 6 patients are alive without recurrence and metastasis. We obtained a good postoperative course in our selected patients with low stage. Thus it was considered that partial nephrectomy is effective against small renal cell carcinoma.

[A case of oligozoospermia with marginal hyperprolactinemia due to pituitary microadenoma].

We present an infertility male with prolactin level of 37.5 ng/ml. The patient had pituitary microadenoma detected by Gd-DTPA enhanced magnetic resonance imaging (MRI). After bromocriptine was administrated for 4 months, the size of microadenoma decreased, and sperm density and mortality improved. His wife became pregnant after 6 months. Dynamic MRI is a useful modality for detection of pituitary microadenoma, and bromocriptine is also useful for treatment of oligospermic patients with marginal hyperprolactinemia.

[Leiomyoma of the bladder: report of two cases].

We report 2 cases of leiomyoma of the urinary bladder. A 41-year-old female visited our hospital with the complaint of pollakisuria. A solid tumor of the urinary bladder was found by ultrasonography. A large shadow defect at the left-anterior wall was shown by drip infusion pyelography (DIP). Computed tomographic scan (CT) and magnetic resonance imaging (MRI) also revealed a large tumor. T1-weighted image revealed a homogeneous low intensity tumor and T2-weighted image disclosed heterogeneous low intensity tumor. Cystoscopy revealed a large submucosal tumor. Partial cystectomy was performed, and she has had neither recurrence nor metastasis for 36 months. A 32-year-old male was referred to our hospital with the complaint of macrohematuria. A solid tumor of the urinary bladder was found by ultrasonography. A shadow defect was not clearly detected by DIP. A large tumor was detected on the anterior wall by MRI. T1-weighted image showed a homogeneous low intensity tumor and T2-weighted image disclosed a high intensity tumor. Cystoscopy revealed a submucosal tumor on the anterior wall. Urine cytology did not suggest a malignancy. The biopsied specimens revealed only an inflammatory change in the mucosa. Partial cystectomy was carried out. He has had neither recurrence nor metastasis for 29 months. Histological diagnosis in both cases was leiomyoma of the urinary bladder.

[Effects of cernitin pollen-extract (Cernilton) on inflammatory cytokines in sex-hormone-induced nonbacterial prostatitis rats].

Cernitin pollen-extract (Cernilton, CN) is a preparation made from eight kinds of pollen and has been used for various prostatic diseases in Japan and Europe. We reported previously that CN possessed a recovery action on the sex-hormone-induced nonbacterial prostatitis in rats. To clarify the possible mechanism of action of CN, we investigated the effects of CN on inflammatory cytokines (IL-1 beta, IL-6 and TNF-alpha) in the same model. Aged Wistar rats were castrated and injected 17 beta-estradiol (0.25 mg/kg/day, s.c.) for 30 days. CN (630 and 1,260 mg/kg, p.o.) or testosterone (2.5 mg/kg, s.c.) was administered for the last 14 days of the treatment of 17 beta-estradiol. In control rats, prostatic IL-6 and TNF-alpha contents were increased approximately 2-3 fold, and acinar glandular inflammation and stromal proliferation were found histopathologically, as compared with those of intact rats. On the other hand, CN decreased the increased contents of cytokines in a dose-dependent manner. The histopathological changes mentioned above were restored in rats treated with 1,260 mg/kg. Testosterone also ameliorated them significantly. These results indicate that CN has an anti-inflammatory action, and that the inhibitory effect of CN on the prostatic inflammatory cytokine is an important factor in its action.

Excessive expression of Txk, a member of the Tec family of tyrosine kinases, contributes to excessive Th1 cytokine production by T lymphocytes in patients with Behcet's disease.

Excessive Th1 cell function is importantly involved in the pathogenesis of Behcet's disease (BD). We previously found that Txk, a member of the Tec family of tyrosine kinases, acts as a Th1 cell specific transcription factor. To investigate immune aberration in the pathogenesis of BD, we studied the expression of Txk and Th1 cytokines in peripheral blood lymphocytes (PBL) and skin lesions in patients with BD. Cytokine production by the lymphocytes was assessed using ELISA. PBL produced excessive Th1 associated cytokines including IFN-gamma and IL-12 spontaneously and in response to exogenous HSP60-derived peptide stimulation, which was shown to induce proliferation of PBL, in patients with BD. Circulating CD4+ T cells expressed excessive Txk protein. A majority of cells infiltrating into skin lesions expressed IFN-gamma in the BD specimens. IL-12 and IL-18 were also expressed in the mononuclear cell aggregates. Lymphocytes accumulating in the skin lesion expressed higher levels of Txk as compared with atopic dermatitis lesions, a typical Th2 disease. IFN-gamma, IL-18 and Il-12 were detected in the BD skin lesions, which may induce preferential development of Th1 cells in patients with BD. The mononuclear cell aggregates contained Txk expressing cells in such skin lesions. Collectively, Txk expressing Th1 cells and the Th1 associated cytokines may play a critical role in the development of skin lesions in BD.