Inducible nitric oxide synthase and tumor necrosis factor-alpha in delayed gastric emptying and gastrointestinal transit induced by lipopolysaccharide in mice.

Gastrointestinal motility disturbances during endotoxemia are probably caused by lipopolysaccharide (LPS)-induced factors: candidates include nitric oxide (NO), tumor necrosis factor-alpha (TNF-alpha), interleukin-1ss, and interleukin-6. Flow cytometry was used to determine the effects of LPS and these factors on gastric emptying (evaluated indirectly by determining percent gastric retention; %GR) and gastrointestinal transit (GIT) in male BALB/c mice (23-28 g). NO (300 microg/mouse, N = 8) and TNF-alpha (2 microg/mouse, N = 7) increased (P < 0.01) GR and delayed GIT, mimicking the effect of LPS (50 microg/mouse). During early endotoxemia (1.5 h after LPS), inhibition of inducible NO synthase (iNOS) by a selective inhibitor, 1400 W (150 microg/mouse, N = 11), but not antibody neutralization of TNF-alpha (200 microg/mouse, N = 11), reversed the increase of GR (%GR 78.8 +/- 3.3 vs 47.2 +/- 7.5%) and the delay of GIT (geometric center 3.7 +/- 0.4 vs 5.6 +/- 0.2). During late endotoxemia (8 h after LPS), both iNOS inhibition (N = 9) and TNF-alpha neutralization (N = 9) reversed the increase of GR (%GR 33.7 +/- 2.0 vs 19.1 +/- 2.6% (1400 W) and 20.1 +/- 2.0% (anti-TNF-alpha)), but only TNF-alpha neutralization reversed the delay of GIT (geometric center 3.9 +/- 0.4 vs 5.9 +/- 0.2). These findings suggest that iNOS, but not TNF-alpha, is associated with delayed gastric emptying and GIT during early endotoxemia and that during late endotoxemia, both factors are associated with delayed gastric emptying, but only TNF-alpha is associated with delayed GIT.

Caudoputamen is damaged by hypocapnia during mechanical ventilation in a rat model of chronic cerebral hypoperfusion.

BACKGROUND AND PURPOSE: Postoperative brain dysfunction, such as delirium, is a common complication of anesthesia and is sometimes prolonged, especially in patients with cerebrovascular disease. In the present study we investigated the effect of hypocapnia during anesthesia on neuronal damage using a rat model of chronic cerebral hypoperfusion. METHODS: Chronic cerebral hypoperfusion was induced by clipping the bilateral common carotid arteries in male Wistar rats. Fourteen days after the operation, these animals were mechanically ventilated for 2 hours and then kept in suitable conditions for an additional 14 days. Twenty-four rats were assigned to 4 groups: those with chronic cerebral hypoperfusion with either hypocapnia or normocapnia during anesthesia, and those given sham operation with either hypocapnia or normocapnia. White matter lesions in the brain sections were evaluated with Klüver-Barrera staining. Proliferation of glial cells was estimated with the use of immunohistochemistry of glial fibrillary acidic protein, a marker for astroglia, and CD11b, a marker for microglia. Computer-assisted morphometry was applied to the immunohistochemical results of microtubule-associated protein 2 to evaluate the loss of neurons. RESULTS: The histological damage was localized almost exclusively in the white matter in the rats subjected to chronic cerebral hypoperfusion but without hypocapnia. Neuronal damage and astroglial proliferation occurred with aggravated white matter lesions in the caudoputamen in the rats with chronic cerebral hypoperfusion and hypocapnia. No lesions were observed in sham-operated rats with either hypocapnia or normocapnia. CONCLUSIONS: These results indicate that hypocapnia during anesthesia causes tissue damage in the caudoputamen, which may be responsible for long-lasting postoperative delirium in patients with stroke and/or dementia.

Cloning and characterization of two novel aldo-keto reductases (AKR1C12 and AKR1C13) from mouse stomach.

In contrast to hepatic hydrosteroid dehydrogenases (HSDs) of the aldo-keto reductase family (AKR1C), little is known about a stomach one. From a mouse stomach cDNA library, we isolated two clones encoding proteins of 323 amino acid residues. They exhibited 93.2% amino acid sequence identity and 64-68% with any known HSDs. Recombinant proteins expressed in Escherichia coli reduced 9,10-phenanthraquinone with NAD(P)H as cofactor. The mRNAs were exclusively expressed in stomach, liver and ileum. The present study demonstrates that these proteins are new members of the HSD subfamily and they are named AKR1C12 and AKR1C13. Immunohistochemical analysis suggests that they are involved in detoxification of xenobiotics in the stomach.

Characterization of nociceptin hyperalgesia and allodynia in conscious mice.

1. Intrathecal (i.t.) administration of nociceptin and high doses of morphine induced allodynia in response to innocuous tactile stimuli, and i.t. nociceptin evoked hyperalgesia in response to noxious thermal stimuli in conscious mice. Here we have characterized the nociceptin-induced allodynia and compared it with the morphine-induced allodynia and the nociceptin-evoked hyperalgesia. 2. Nociceptin-induced allodynia was evoked by the first stimulus 5 min after i.t. injection, reached a maximum at 10 min, and continued for a 50 min experimental period. Dose-dependency of the allodynia showed a bell-shaped pattern from 50 pg to 5 ng kg-1, and the maximum effect was observed at 2.5 ng kg-1. 3. Morphine-induced allodynia reached the maximum effect at 15 min and declined progressively until cessation by 40-50 min. The dose-response curve showed a bell-shaped pattern, similar to that induced by nociceptin, with a maximum effect at 0.5 mg kg-1, five orders of magnitude higher than that of nociceptin. 4. The allodynia evoked by nociceptin and morphine were dose-dependently blocked by glycine, D(-)-2-amino-5-phosphonovaleric acid (D-AP5, an N-methyl-D-aspartate (NMDA) receptor antagonist), gamma-D-glutamylaminomethyl sulphonic acid (GAMS, a non-NMDA receptor antagonist) and methylene blue (a soluble guanylate cyclase inhibitor), but were not affected by muscimol (a gamma-aminobutyric acidA (GABAA) receptor agonist) and baclofen (a GABAB receptor agonist). 5. Morphine did not inhibit forskolin-stimulated cyclicAMP formation in cultured cells expressing the nociceptin receptor. 6. Nociceptin-induced hyperalgesia was evoked 10-15 min after i.t. injection. Nociceptin produced a monophasic hyperalgesic action over a wide range of doses from 5 fg to 50 ng kg-1. The nociceptin-induced hyperalgesia was blocked by glycine only among the agents examined. 7. None of the pain responses evoked by nociceptin and morphine were blocked by naloxone. 8. These results demonstrate that, whereas the mechanisms of the nociceptin-induced allodynia and hyperalgesia are evidently distinct, they involve a common neurochemical event beginning with the disinhibition of the inhibitory glycinergic response. Morphine may induce allodynia through a pathway common to nociceptin, but the nociceptin receptor does not mediate the action of high doses of morphine.

Novel V184E MEN1 germline mutation in a Japanese kindred with familial hyperparathyroidism.

We studied the MEN1 gene in a kindred where three patients (the proposita and two of her sons) were affected with hyperparathyroidism. By polymerase chain reaction (PCR)-based direct sequencing of 10 exons of MEN1, a novel germline mutation was identified in the proposita. This mutation, a T-to-A transition at codon 184 in exon 3, predicts an amino acid change from valine to glutamine (V184E). PCR-single-strand conformational polymorphism (PCR-SSCP) analysis of exon 3 followed by sequencing showed the same mutation in the two sons, and in two clinically normal granddaughters of an affected son. Since the T-to-A substitution segregated with the disorder in the kindred except for the granddaughters and it was not detected in 100 alleles from 50 normal individuals, the change observed in MEN1 is not a polymorphism, but causes familial hyperparathyroidism. Thus the two grandchildren with the mutation were diagnosed as presymptomatic carriers.

Clinical evaluation of upper mediastinal dissection for differentiated thyroid carcinoma.

BACKGROUND: An extensive upper mediastinal dissection in advanced differentiated thyroid carcinoma is occasionally required. This investigation was undertaken to clarify the indications for mediastinal lymph node dissection and the route of upper mediastinal metastases. METHODS: Twenty-one patients with differentiated thyroid cancer, who underwent their first radical operations with mediastinal dissection through a partial midline sternotomy, were enrolled in this study. Of 21 patients, 10 (48%) were found to have mediastinal lymph node metastases. RESULTS: The tumor size in the group with metastatic disease was much bigger than that in the group without metastatic disease. Histologic type and age were similar between the two groups. The extent of cervical lymph node metastases was more significant in the group with metastatic disease; in particular, all 10 patients showed more than two metastatic nodes along the internal jugular vein of the tumor-free side. CONCLUSIONS: This study indicates that metastases to the internal jugular chain on the side contralateral to the primary tumor would be an extremely important factor for indication of extensive upper mediastinal lymph node dissection after median partial sternotomy in patients with differentiated thyroid carcinoma.

High intra-abdominal pressure increases plasma catecholamine concentrations during pneumoperitoneum for laparoscopic procedures.

BACKGROUND: Laparoscopic procedures are associated with several complications, such as hemodynamic, respiratory, and endocrine complications. In our previous clinical study, plasma epinephrine and norepinephrine concentrations remained unchanged after the insertion of a Veress needle, but increased significantly immediately after insufflation with carbon dioxide into the peritoneum. The mechanisms for this increase are unknown. OBJECTIVE: To investigate whether gas insufflation during pneumoperitoneum affects plasma catecholamine concentrations during laparoscopic procedures. DESIGN: Experimental study in pigs. MAIN OUTCOME MEASURES: The plasma concentrations of epinephrine and norepinephrine were measured in the pigs before and after pneumoperitoneum. The mean arterial pressure, heart rate, cardiac output, and arterial blood gas levels were measured, and the systemic vascular resistance was calculated. INTERVENTION: Air, nitrous oxide, or carbon dioxide were insufflated in turn into the peritoneal cavity of supine pigs. Thereafter, carbon dioxide was insufflated into the peritoneal cavity while the pig was in the left lateral decubitus position, and then in the right lateral decubitus position. Measures were performed before pneumoperitoneum and at the intra-abdominal pressures of 10 mm Hg and 20 mm Hg. One hour of resting time was allowed between each procedure. RESULTS: As compared with baseline values, the plasma concentrations of epinephrine and norepinephrine remained unchanged at 10 mm Hg but increased significantly at 20 mm Hg regardless of the gas used for the pneumoperitoneum (P<.05). The type of gas and differences in the position of the animals had no effect on the plasma epinephrine and norepinephrine concentrations. CONCLUSIONS: Excessive intra-abdominal pressure, but not the type of gas or body position, increases plasma catecholamine concentrations during the insufflation of gas into the abdominal cavity. Therefore, excessive insufflation of the pneumoperitoneum should be avoided.

The effect of ketamine isomers on both mice behavioral responses and c-Fos expression in the posterior cingulate and retrosplenial cortices.

Ketamine, a non-competitive NMDA receptor antagonist, is a racemic mixture. S(+) ketamine is presumed to be more potent as an anesthetic than R(-) ketamine, and causes less postanesthetic stimulation of locomotor activity than R(-) ketamine in animals at equihypnotic doses. In the present study, we investigated the effect of S(+), R(-), and racemic ketamines on mice behavioral responses and c-Fos expression in the posterior cingulate and retrosplenial cortices (PC/RS), which are suggested to be the brain regions responsible for NMDA-receptor-antagonist-induced psychotomimetic activity. Ataxia and head weaving and c-Fos expression in the PC/RS were significantly more induced by both S(+) and racemic ketamines than by R(-) ketamine at the same dose. S(+) ketamine induced significantly more potent ataxia than racemic ketamine at the same dose. Ketamine-induced c-Fos expression in the PC/RS correlated well with the intensity of behavioral responses. These results imply that R(-) ketamine is weaker than both S(+) and racemic ketamines in a psychotomimetic effect. Also, S(+) ketamine is more potent than racemic ketamine in a psychotomimetic effect and possibly in an anesthetic effect. They also indicate that PC/RS is at least one of the specific brain regions responsible for ketamine-induced behavioral responses in animals and a psychotomimetic activity in humans.

Significance of values of thyrotropin binding inhibitor immunoglobulins and appearance of intrathyroidal lymphocytes at subtotal thyroidectomy for Graves' disease.

This study was undertaken to investigate whether thyrotropin binding inhibitor immunoglobulins (TBII) values at and after subtotal thyroidectomy correlated with the outcome or the histologic grade applied to the appearance of intrathyroidal lymphocytes at the time of surgical treatment. In addition, the main reason for the study was to determine whether or not it was possible to predict the outcome after the operation by the data obtained. There was no relation between the TBII results and the grade of the appearance of intrathyroidal lymphocytes at the time of operation or the TBII postoperative results (whether positive or negative) and the final outcome. However, it was of interest that patients with a recurrence of hyperthyroidism had the TBII values of more than 50 percent at the time of surgical treatment, and also manifested continuously positive TBII values after the operation. They also had moderate grades of lymphocytic infiltration and lymph follicle formation in the surgical specimen. It seemed impossible to predict the outcome of each instance in accordance with TBII values and the grade of the appearance of intrathyroidal lymphocytes at the time of the operation. However, it might be possible to predict at least the recurrence of hyperthyroidism by the consideration of changes of TBII values postoperatively.

Ergostane glycosides from Petunia hybrida.

Six new ergostane glycosides, designated as petunioside A, petunioside B, 24-epipetunioside B, petunioside C, 24-epipetunioside C and petunioside D, were isolated from the methanolic extract of the fresh aerial parts of Petunia hybrida. Their structures were determined by means of spectroscopic analysis and single crystal X-ray analysis.

Clinical significance of the increased multidrug resistance-associated protein (MRP) gene expression in patients with primary breast cancer.

The efficacy of cancer treatment is limited by either intrinsic or acquired resistance to various chemotherapeutic agents. To evaluate the clinically important factors related to prognosis in primary breast cancer retrospectively, we investigated the expression of the following genes involving acquirement of drug resistance: multidrug resistance 1 (MDRl), multidrug resistance-associated protein (MRP), and topoisomerase (Topo) I, II alpha, and II beta. Using an RT-PCR method, we semiquantified the gene expression level in untreated stage II breast cancer tissue (n = 27) and noncancerous breast tissue (n = 10). Among the 27 cancer patients, who were all treated by adjuvant chemoendocrine therapy after surgery, 10 patients showed relapse within the following 10 years whereas 17 patients did not. The gene expression levels of MDRl, MRP, and Topo I, II alpha, and II beta were normalized to the level of the beta 2-microglobulin RT-PCR product. MRP mRNA expression was detected in 70% of the breast cancer tissues and its expression levels were significantly increased in the cancer group compared with the noncancerous breast tissues. Furthermore, the MRP level was much higher in the relapsed patient group. On the other hand, there were no significant differences in the MDRl mRNA levels between the noncancerous and cancer groups. Although Topo II alpha mRNA was not detected in noncancerous breast tissues, it was detected in 52% of the breast cancer tissues. In cancer patients, no significant difference in Topo II alpha mRNA levels was observed between the relapsed and nonrelapsed groups. These findings suggest that MRP might be used as one of the markers for poor prognosis in patients with breast cancer.

Three-dimensional structure of the micro-blood vessels in thyroid tumors analyzed by immunohistochemistry coupled with image analysis.

The structure of micro-blood vessels, one of the most important factors influencing the tumor growth and tumor metastasis among histological types of thyroid malignancy, was analyzed immunochemically by staining tissues for platelet endothelial cell adhesion molecule-1 (PECAM-1). Human thyroid tumor tissue obtained at surgery, consisting of 18 cases of papillary carcinoma, 9 cases of follicular carcinoma, and 9 cases of anaplastic carcinoma were fixed in formalin solution, and paraffin sections were made. They were stained for PECAM-1 using the avidin-biotin complex (ABC) technique. The volume of the blood vessels and their three-dimensional (3D) structure were analyzed using an image analyzer. The volume ratios of blood vessels in thyroid tissues were: normal tissues, 1.10%; papillary carcinoma, 3.01%; follicular carcinoma, 8.13%; and anaplastic carcinoma, 0.91%. Ratios in malignant tumors were larger than in normal tissues, except for anaplastic carcinoma. The typical 3D structure of micro-blood vessels was histopathologically varied: branching tree-like blood vessels in papillary carcinomas; vessels of varied diameter surrounding follicle structure in follicular carcinomas; and simple and immature vessels in anaplastic carcinomas. The volume and 3D structure of micro-blood vessels in thyroid malignant tumors differed from those in normal tissues, and varied according to histological classification.

Effectiveness of preoperative radioactive iodine (131I) therapy for locally advanced papillary thyroid cancer: a case report.

We report here the effectiveness of preoperative radioactive iodine-131 (131I) therapy for locally advanced thyroid cancer. A 57-year-old woman demonstrated a hard neck tumor that markedly invaded the surrounding organs. The cytological diagnosis of the tumor using fine-needle aspiration biopsy was papillary carcinoma. Because curative resection of the tumor appeared difficult at her first visit, 131I therapy was performed prior to surgery and was more useful than expected. After 3 131I treatments, the tumor size was greatly reduced, and the patient underwent a curative operation. Histopathological diagnosis was well differentiated papillary carcinoma, pT4 and pN1b. The postoperative clinical course was uneventful. There have been no definitive reports using 131I as preoperative treatment for inoperable thyroid cancer. We suggest that 131I therapy may also be beneficial as neoadjuvant therapy for locally advanced thyroid carcinoma.

Increased chromatid-type chromosomal aberrations in mouse m5S cells exposed to power-line frequency magnetic fields.

PURPOSE: To investigate the induction of chromosomal aberrations in mouse m5S cells after exposure to power-line frequency magnetic fields (extremely low frequency magnetic fields; ELFMF) at high-flux densities. MATERIAL AND METHOD: m5S cells were either untreated or pretreated during the G1 phase with mitomycin C (MMC, 1 microM) for 1 h or 3 Gy X-rays, and then exposed to ELFMF at three different flux densities (5 and 50 mT at 60 Hz, 400 mT at 50 Hz) for 40 h. Unexposed control cells were incubated for the same period in a conventional CO2 incubator. Chromosomal aberrations were analysed in the first post-treatment metaphases. Cell kinetics were assessed by DNA flow cytometry and the mitotic index. RESULTS AND CONCLUSIONS: ELFMF enhanced the formation of spontaneous and MMC- or X-ray-induced chromosomal aberrations, in a flux-density-dependent manner. Statistically significant increases in the frequency of chromosomal aberrations were observed in cells exposed to 400 mT ELFMF with respect to unexposed controls. The aberrations induced by ELFMF were mostly chromatid-type, not chromosome-type. The cells exposed to 400 mT ELFMF exhibited a three-fold higher level of chromatid-type aberrations than did the unexposed cells. Flow cytometric and mitotic index analyses revealed that the S or G2 arrest following MMC or X-irradiation was more profound in ELFMF-exposed cells than in unexposed cells. Our results suggest that ELFMF can interfere with post-replication repair, resulting in increased levels of chromatid-type chromosomal aberrations induced spontaneously and by DNA damaging agents.

Microcapsulated parathyroid tissue in vitro.

Ideally, microcapsules will make it possible to transplant parathyroid tissue for allo- and xenotransplantation in hypoparathyroid patients. In this study, parathyroid tissues capsulated by polyelectrolyte complex were evaluated in vitro. Hormone secretion ability was significantly (P < 0.001) greater in sediment than in supernatant. There was no difference in hormone secretion ability between the parathyroid tissue thawed at room temperature and the tissue thawed in a bath at 37 degrees C. Both non-capsulated and capsulated parathyroid tissues were incubated for three weeks. The ability of capsulated parathyroid to secrete hormones in vitro was reduced gradually and disappeared within three weeks. There are still several problems to be solved before the clinical application of parathyroid allotransplantation using microcapsulation.

Bilateral synchronous spindle cell, apocrine and scirrhous carcinoma breast cancer in a case or malignant lymphoma.

An unusual case of synchronous bilateral breast cancer occurring during combination chemotherapy and radiation to the outside of the breast for malignant lymphoma is reported. Two histologically rare carcinomas, spindle cell carcinoma and apocrine carcinoma, were observed in this case. A 77-year-old woman, who had been treated for stage IIIA non-Hodgkin's lymphoma, developed bilateral breast tumors. Aspiration biopsy cytology findings of the tumor in the left breast showed several clusters of adenocarcinoma cells and some large atypical spindle shaped cells, which suggested spindle cell carcinoma. The cytologic findings of the right breast tumor were highly suggestive of scirrhous carcinoma. A modified radical mastectomy was performed on both breasts. Pathological examination disclosed two separate cancer lesions in the left breast. The lesion which had been detected before the operation, was a spindle cell carcinoma. Another lesion, detected for the first time by pathological examination, was an apocrine carcinoma. The lesion in the right breast was a scirrhous carcinoma. Since non-invasive foci were detected in these three cancer lesions, each lesion was thought to be a primary cancer. All dissected bilateral axillary lymph nodes showed malignant lymphoma. Immuno-histochemistry of the spindle cell carcinoma revealed positive immunoreactivity for cytokeratin, which suggested the epithelial as well as mesenchymal nature of this tumor. Synchronous existence of malignant lymphoma and three independent breast cancers including spindle cell carcinoma and apocrine carcinoma is very rare.

Hoarseness after cardiac surgery.

OBJECTIVE: To investigate the incidence, severity, and duration of hoarseness after adult cardiac surgery and to examine movement of the endotracheal tube (ETT) during neck extension for sternotomy. DESIGN: Prospective study. SETTING: University hospital. PATIENTS: Seventy-two adults undergoing elective cardiac surgery (assessment of hoarseness was completed in 68). INTERVENTIONS: An ETT with an internal diameter of 8.0 mm and 7.5 mm was used for men and women, respectively. After surgical positioning, the ETT was placed with its tip about 3 cm above the carina. MEASURES: ETT migration during positioning for sternotomy was measured using a fiberoptic bronchoscope. Hoarseness was examined at 6-12 hrs after extubation. RESULTS: The ETT moved away from the carina during surgical positioning and the migration was 11+/-6 (1-27) mm [mean+/-SD (range)]. Hoarseness occurred in 32% of the patients (22/68), of whom five (23%) were grade 1 on a four-point scale (0-3), 14 (64%) were grade 2, and three (14%) were grade 3 (aphonic). The duration of hoarseness was 4+/-3 (1-14) days, except in one patient with left vocal cord paralysis (60 days). When this patient was excluded, intubation time was an independent predictor of the severity of hoarseness (p=0.0001). CONCLUSIONS: Hoarseness after cardiac surgery is common. Most hoarseness is transient and its severity is predicted by duration of intubation. Positioning for sternotomy displaces the ETT cuff away from the carina. This may cause persistent hoarseness because displaced cuff from trachea to larynx may damage the recurrent laryngeal nerve or vocal cords.

Secondary malignant lymphoma which simulated primary thyroid cancer.

A patient with secondary thyroid lymphoma who complained of a neck mass was presented. Multiple nodules were detected in both lobes of the thyroid gland, which appeared as homogeneous hypoechoic masses on ultrasonogarphy (US), low-density masses on computed tomography (CT), and areas of increased uptake on gallium-67 scintigraphy. The residual thyroid gland was normal. Surgery established a diagnosis of secondary thyroid lymphoma with no coexistent Hashimoto's thyroiditis but with cervical node involvement by lymphoma. Radiologic evidence of normal residual thyroid gland in a patient with thyroid lymphoma may be a sign of secondary thyroid lymphoma.