Intraoperative administration of inhaled carbon monoxide reduces delayed graft function in kidney allografts in Swine.

Ischemia/reperfusion injury and delayed graft function (DGF) following organ transplantation adversely affect graft function and survival. A large animal model has not been characterized. We developed a pig kidney allograft model of DGF and evaluated the cytoprotective effects of inhaled carbon monoxide (CO). We demonstrate that donor warm ischemia time is a critical determinant of DGF as evidenced by a transient (4-6 days) increase in serum creatinine and blood urea nitrogen following transplantation before returning to baseline. CO administered to recipients intraoperatively for 1 h restored kidney function more rapidly versus air-treated controls. CO reduced acute tubular necrosis, apoptosis, tissue factor expression and P-selectin expression and enhanced proliferative repair as measured by phosphorylation of retinol binding protein and histone H3. Gene microarray analyses with confirmatory PCR of biopsy specimens showed that CO blocked proinflammatory gene expression of MCP-1 and heat shock proteins. In vitro in pig renal epithelial cells, CO blocks anoxia-reoxygenation-induced cell death while promoting proliferation. This large animal model of DGF can be utilized for testing therapeutic strategies to reduce or prevent DGF in humans. The efficacy of CO on improving graft function posttransplant validates the model and offers a potentially important therapeutic strategy to improve transplant outcomes.

Total tumour volume as a prognostic factor in patients with resectable colorectal cancer liver metastases.

BACKGROUND: Although total tumour volume (TTV) may have prognostic value for hepatic resection in certain solid cancers, its importance in colorectal liver metastases (CRLM) remains unexplored. This study investigated its prognostic value in patients with resectable CRLM. METHOD: This was a retrospective review of patients who underwent hepatic resection for CRLM between 2008 and 2017 in a single institution. TTV was measured from CT images using three-dimensional construction software; cut-off values were determined using receiver operating characteristic (ROC) curve analyses. Potential prognostic factors, overall survival (OS) and recurrence-free survival (RFS) were determined using multivariable and Kaplan-Meier analyses. RESULTS: Some 94 patients were included. TTV cut-off values for OS and RFS were 100 and 10 ml respectively. Right colonic primary tumours, primary lymph node metastasis and bilobar liver metastasis were included in the multivariable analysis of OS; a TTV of 100 ml or above was independently associated with poorer OS (hazard ratio (HR) 6·34, 95 per cent c.i. 2·08 to 17·90; P = 0·002). Right colonic primary tumours and primary lymph node metastasis were included in the RFS analysis; a TTV of 10 ml or more independently predicted poorer RFS (HR 1·90, 1·12 to 3·57; P = 0·017). The 5-year OS rate for a TTV of 100 ml or more was 41 per cent, compared with 67 per cent for a TTV below 100 ml (P = 0·006). Corresponding RFS rates with TTV of 10 ml or more, or less than 10 ml, were 14 and 58 per cent respectively (P = 0·009). A TTV of at least 100 ml conferred a higher rate of unresectable initial recurrences (12 of 15, 80 per cent) after initial hepatic resection. CONCLUSION: TTV was associated with RFS and OS after initial hepatic resection for CRLM; TTV of 100 ml or above was associated with a higher rate of unresectable recurrence.

ANTECEDENTES: Aunque el volumen total del tumor (total tumour volume, TTV) puede tener valor pronóstico tras la resección hepática (hepatic resection, HR) en algunas neoplasias sólidas, no se conoce su importancia en las metástasis hepáticas de cáncer colorrectal (colorectal liver metastases, CRLMs). Este estudio analizó el valor pronóstico del TTV en pacientes con CRLMs resecables. MÉTODOS: Revisión retrospectiva de pacientes a los que se realizó una HR por CRLMs entre 2008 y 2017 en un solo centro. El TTV se estimó a partir de imágenes de tomografía computarizada utilizando un programa de reconstrucción 3D; se determinaron los valores de corte mediante un análisis de las características operativas del receptor. Se identificaron los posibles factores pronósticos y se calcularon la supervivencia global (overall survival, OS) y la supervivencia libre de recidiva (recurence-free survival, RFS) mediante análisis multivariados y de Kaplan-Meier. RESULTADOS: Se incluyeron 94 pacientes. Los valores de corte del TTV para la OS y la RFS fueron de 100 mL y 10 mL, respectivamente. En el análisis multivariable para la OS, se incluyeron los tumores del colon derecho, las metástasis linfáticas primarias y la metástasis hepática bilobar; un TTV ≥ 100 mL se asoció de forma independiente con una peor OS (cociente de riesgos instantáneos, hazard ratio, HR, 6,34, i.c. del 95% 2,08-17,9; P = 0,002). En el anáisis para la RFS, se incluyeron tumores primarios de colon derecho y las metástasis linfáticas primarias; un TTV ≥ 10 mL predijo de forma independiente una peor RFS (HR 1,90, i.c. del 95% 1,12-3,57; P = 0,017). Las tasas de OS a los 5 años con TTVs ≥ 100 mL versus < 100 mL fueron del 41% versus 67% (P = 0,006); las tasas de RFS respecto a TTVs ≥ 10 mL versus < 10 mL fueron del 14% versus 58% (P = 0,009). Un TTV ≥ 100 mL conllevó una tasa más elevada (80%) de recidivas no resecables después de la HR inicial. CONCLUSIÓN: El TTV se asoció con la RFS y la OS tras la HR por CRLMs; unos valores ≥ 100 mL conllevan una tasa más elevada de recidiva irresecable.

Effect of rush immunotherapy in house-dust-mite (HDM)-sensitive adult bronchial asthma: changes in in vivo and in vitro responses to HDM.

An open study was conducted to evaluate the changes in in vivo and in vitro responses to house-dust-mite (HDM) after rush immunotherapy (RI). A 7-day RI protocol using an extract containing HDM allergen was administered to 12 subjects with HDM-sensitive asthma, and the effects on bronchial responsiveness and serum antibody levels were evaluated up to 16 or 20 weeks after RI. The levels of HDM-specific IgG, IgG1 and IgG4 antibodies were significantly elevated from 4 or 8 weeks after RI. Provocative doses causing a 20% fall in FEV1 (PD20) by allergen inhalation were elevated in all subjects at 16 to 20 weeks after RI. There was a high correlation between the increase in log-PD20 and the increase in the ratio of HDM-specific IgG4 to IgG1 (r = 0.68, p < 0.05). The results suggest that RI elicits the improvement of allergen-specific bronchial responsiveness and the increase in serum antibody levels within a relatively short period.

Immunohistochemical microquantification of fast-myosin in frozen histological sections of mammalian skeletal muscles.

Fast-myosin in frozen histological sections was quantified by an immunohistochemical micromethod based on the ELISA. Frozen tissue sections mounted on glass slides were used analogously to the antigen-precoated wells of ELISA plates. The intensity of immunoreactivity of frozen sections to an anti-fast-myosin monoclonal antibody was quantified directly from the color developed with the second antibody coupled with peroxidase using phenol-4-aminoantipyrine as a substrate. Fast-myosin levels in the masseter muscles of pigs, rats, and rabbits were 185 +/- 6, 223 +/- 9, and 178 +/- 12 mg/g of total protein, respectively, and those in the gastrocnemius muscles from cows, pigs, goats, rats, and rabbits were 172 +/- 12, 211 +/- 7, 177 +/- 9, 211 +/- 10, and 205 +/- 10 mg/g, respectively. In the masseter of cows and goats, fast-myosin was not detected. The results obtained by this immunohistochemical micromethod were in good agreement with those obtained by histomorphometrical and biochemical analyses. This immunohistochemical micromethod could be used to quantitatively evaluate the muscle contractile characteristics that determine meat quality.

Characteristics of lectin staining patterns assessed by a modified sensitive thermo-method in rat livers with heterologous serum-induced fibrosis.

Lectins are sensitive probes which bind carbohydrate structures specifically. In this study, we modified the lectin staining procedure for sensitive detection of carbohydrate structures in formalin-fixed, paraffin-embedded sections of normal and heterologous serum-induced fibrotic livers. The liver sections were heated in hot distilled water at 100 degrees C for 10 min (thermo-treatment: TT), and then stained with 24 different lectins. In comparison with the results from sections without TT (nonTT), enhanced and/or alternated staining patterns of 19 lectins were demonstrated in sections with TT, and enhanced staining of Vicia villosa agglutinin seen in Kupffer cells was noted. Interestingly, no positive staining was seen with Dolichos biflorus agglutinin, peanut agglutinin or soybean agglutinin (SBA), which recognize O-linked carbohydrate chains, in Kupffer cells of non-TT sections, but strong positive staining was demonstrated in those of TT sections. SBA-positive staining in the cytoplasm of some scattered hepatocytes located in the periportal and perifibrous zones and central zone of pseudolobules was demonstrated only in the fibrotic liver sections with TT. Such findings indicate the heterogeneity of hepatocytes in the liver with fibrosis. Formalin fixation causes masking of lectin binding sites, especially O-linked carbohydrate chains, and TT may recover such masking reactions. TT improved the staining reactions for many lectins in formalin-fixed, paraffin-embedded liver sections, and new staining patterns appear after TT. Modified TT staining procedures may be useful for the diagnosis and prognosis of liver fibrosis.

Lectin histochemistry in rat liver fibrosis induced by heterologous serum sensitization.

The localization of carbohydrates in rat livers with fibrosis induced by heterologous serum was examined by lectin histochemical and biochemical techniques. Twenty-four lectins were used to visualize the different carbohydrates in paraffin sections of normal and fibrotic liver tissues. No differences in staining patterns of these lectins were observed between normal and fibrotic livers in hepatocyte cell membranes including bile canaliculi, sinusoidal endothelial, or bile ductal cells. Kupffer cells strongly stained with Vicia villosa agglutinin (VVA) were seen only in the periportal zone of the normal liver, but they were observed in the periportal zone and scattered throughout the pseudolobular zone in the fibrotic liver. The cytoplasm of some hepatocytes was strongly stained by Bandeiraea simplicifolia lectin-I (BSL-I). BSL-I positive hepatocytes in normal liver were localized in the periportal zone, but those in the fibrotic liver were scattered in the periportal and perifibrous zones. After polyacrylamide gel electrophoresis of liver glycoproteins, differences in molecular sizes of BSL-I positive glycoproteins (79 and 81 kD) were detected by lectin blotting. Cell density of perifibrous BSL-I positive hepatocytes may be useful as a diagnostic parameter for liver fibrosis and/or cirrhosis. Two distinct staining patterns with twelve lectins were observed in fibrotic septa of the fibrotic liver. The fibrotic septa were stained with six characteristic lectins, and the centrilobular septa were stained with all these twelve of lectins. Histopathological assessment of the centrilobular fibrotic septa stained with these characteristic lectins may contribute to the diagnosis and prognosis of hepatic fibrosis.

Changes in quantitative profile of extracellular matrix components in the kidneys of rats with adriamycin-induced nephropathy.

Extracellular matrix components (ECMs) in histological sections of the kidney cortex from the rats with adriamycin (ADR)-induced nephropathy (5 mg/kg, i.v.) were quantified by an immunohistochemical micromethod. Changes in kidney histopathology and urine and blood biochemistry were investigated. Enlarged kidneys were granular on the surface and pale in color in ADR-treated rats, and these rats had kidneys with glomeruli with expanded mesangial area and with capillary aneurysm. Severe albuminuria, hypoalbuminemia, hypercholesterolemia and disorders in other nephrotic parameters were observed in ADR-treated rats. Type I and IV collagens, fibronectin and laminin contents in the renal cortex of ADR-treated rats at 10 weeks were 329, 317, 263 and 295%, respectively, higher than in each vehicle control, and those at 28 weeks were 1,211, 930, 1,057 and 1,012%, respectively. The glomerular sclerotic abnormalities progressed in a time-dependent manner. Moreover, there was a strong correlation between the ECM levels and serum creatinine and blood urea nitrogen levels. In conclusion, microquantification provided useful information for accurate diagnosis and prognosis of nephrotic lesions and is a good tool to assess the advancement of renal disorders in patients with nephropathy.

[The long-term effect of high-dose beclomethasone dipropionate on the pituitary-adrenal function].

The pituitary-adrenal function was studied in seven asthmatic subjects who had been received daily inhalations of 800 to 1,000 micrograms of beclomethasone dipropionate (BDP) over a year. None of the subjects had taken oral corticosteroids for at least six months prior to the study. As indices of pituitary-adrenal function, 1) circadian rhythm of plasma ACTH and cortisol, 2) urine 17-OHCS and 17-KS, and 3) the response of cortisol in rapid ACTH test were examined. All subjects showed normal circadian rhythms of plasma ACTH and cortisol levels. Urinary 17-OHCS and 17-KS excretions over a 24-hour period were also within the normal range. Plasma cortisol levels in the rapid ACTH test were significantly increased and judged as normal responses in all subjects. These results indicate that long-term treatment with BDP ranging from 800 to 1,000 micrograms/day induces no suppressive effect on the pituitary-adrenal function.

[Clinical evaluation of urinary leukotriene E4 levels in asymptomatic bronchial asthma].

To evaluate the significance of peptide leukotrienes (LTC4, D4, E4) in asymptomatic asthmatic patients, we measured urinary LTE4 levels which is thought to reflect in vivo production of peptide LTs. Urinary LTE4, was extracted using C18 solid phase column and measured by radioimmunoassay. There was no significant difference in urinary LTE4 levels among asthmatics with different severity or between atopic and non-atopic asthmatics. Urinary LTE4 levels were significantly elevated in asthmatics compared with normal controls (p < 0.05). When compared with normal controls, urinary LTE4 levels were significantly elevated in moderate to severe asthma (p < 0.05), and non-atopic asthmatics (p < 0.001). Urinary LTE4 levels were significantly elevated in aspirin-sensitive asthmatics compared with aspirin-tolerant asthmatics (p < 0.05). There was no significant difference in urinary LTE4 levels among aspirin-sensitive asthmatics with different severity. These results suggest that increased production of peptide LTs is a characteristic in aspirin-sensitive asthma, and that the severity and type of asthma and the presence of aspirin-sensitive asthma should be taken into consideration in the analysis of urinary LTE4 levels.

[Combination chemotherapy with bronchial arterial infusion (BAI) and systemic chemotherapy in advanced non-small cell lung cancer].

Five patients of advanced non-small cell lung cancer with atelectasis and/or poor performance status were treated with combination MVP therapy consisting of bronchial arterial infusion (BAI) using cisplatin (50 mg) and simultaneous systemic MVP therapy. The tumor size reduced more than 50% in three patients, and less than 25% in one patient. In another patient the tumor size was not measurable. Four of five patients showed disappearance of atelectasis, and three of four patients showed improvement of performance status. Average time taken was 13 and 17 days, respectively. We expect that combination MVP therapy has beneficial effects not only for relieving local symptoms and improving performance status within shorter period, but also achieving a better prognosis.

[Comparative study on quality of life between weekly and monthly chemotherapy with cisplatin, vindesine and mitomycin C in patients with non-small cell lung cancer].

A comparative study between weekly and monthly chemotherapy with CDDP, VDS and MMC was performed on the basis of QOL in patients (stage III A/III B/IV) with non-small lung cancer from September 1993 to August 1996. Arm A received CDDP (80 mg/m2) monthly on day 1, VDS (3 mg/m2) on days 1 and 8, MMC (8 mg/m2) on day 1. Arm B received CDDP (40 mg/m2), VDS (3 mg/m2) and MMC (4 mg/m2) on day 1, and followed by CDDP (20 mg/m2), VDS (1.5 mg/m2) and MMC (2 mg/m2) weekly. A diary-type QOL self-rating questionnaire was devised for this study, consisting of 5 scales including 13-item questionnaires and a face scale as the global scale. Chronological QOL data from 20 days during chemotherapy (total dose of CDDP was 80 mg/m2 in both A and B arm) were analyzed using summary measures. In the 78 eligible subjects of the study, 34 were QOL subjects of the 27 eligible subjects, 13 were in arm A and 14 in arm B. 1. The questionnaire was confirmed to be satisfactory with regard to reliability and validity as a questionnaire for the evaluation of this study by factor and correlation analysis. 2. Although each scale changed to the worth level after chemotherapy, there was no significant difference between arm A and B. 3. Summary statistics were assessed using indices of the area under the curve (AUC) and the maximum fluctuations of QOL scores (Dif max) arm B showed a tendency to smaller changes than arm A in AUC and Dif max. There was a significant difference in the physical well-being scale of Dif max. 4. Three items, including the physical well-being in arm B, revealed a better score than those of arm A. Abdominal condition showed a significant difference between the two arms. 5. No obvious difference in anti-cancer effectiveness was found between arm A and B. Arm B showed a longer median survival, less nausea and vomiting, leukopenia and thrombocytopenia compared with arm A. The above results suggested that weekly chemotherapy is as effective, less toxic and shows less influence against patients QOL than the conventional monthly chemotherapy in this study.

[Idiopathic pulmonary fibrosis of the upper lobe: a case report].

A 26-year-old man was admitted to our hospital complaining of exertional dyspnea. Chest x-ray films disclosed reticulonodular shadows predominantly in the upper fields of both lungs, but no apical cap. Lung biopsy specimens obtained from the upper lobe by video-assisted thoracoscopy revealed subpleural elasto-fibrosis. Also, specimens obtained from the lower lobes disclosed micro-honeycombing due to peri-lobular fibrosis resembling usual interstitial pneumonia. Although pneumothorax occurred repeatedly, the lungs reinflated on each occasion without artificial intervention. Similar radiographic findings had been obtained on the patient's father, who died of idiopathic pulmonary fibrosis at the age of 56. Idiopathic pulmonary upper lobe fibrosis was conclusively diagnosed because the patient exhibited most of the features originally described by Amitani et al.

Long-term complications after liver transplantation.

BACKGROUND: Along with an increased number of cases of liver transplantation (LT), perioperative mortality has decreased and short-term survival has improved. However, long-term complications have not been fully elucidated today. PURPOSE: Chronic complications were analyzed individually to find risk factors and to improve long-term outcomes after LT. SUBJECTS: There were 63 cases of LT from our outpatient clinic that were included in this study. Among them, 58 were performed using living donor LT and 5 were performed using deceased donor LT. Original diseases mainly consisted of hepatitis C virus (HCV; 45.9%) and hepatitis B virus (23.0%). FINDINGS: The median follow-up was 5.4 ± 3.3 years (range, 0.1∼17 years). Overall survival at 2, 3, 5, and 10 years was 89.3%, 83.4%, 81.3%, and 81.3%, respectively. Long-term complications mainly consisted of renal dysfunction (62.7%), dyslipidemia (29.4%), diabetes mellitus (21.6%), and arterial hypertension (21.6%). In univariate analysis, HCV (P = .03) and elapsed years after LT (P = .02) were identified as predictive factors for arterial hypertension and recipient age >50 (P = .03), and elapsed years after LT for renal dysfunction (P = .03), respectively. In multivariate Cox regression analysis, HCV (odds ratio [OR] 5.25, 95% confidence interval [CI] 1.05-34.06, P = .04) was identified as a predictive factor for arterial hypertension, and recipient age older than 50 years for renal dysfunction (OR 5.67, 95% CI 1.34-28.88, P = .02). The number of elapsed years after transplantation was also identified as a predictive factor for arterial hypertension/dyslipidemia/renal dysfunction (OR 13.88/14.15/4.10, 95% CI 1.91-298.26/2.18-290.78/1.09-18.03, P = .01/.003/.04). Fifty percent of the recipients developed renal dysfunction within 8 years after LT, and fluctuation of estimated glomerular filtration rate (eGFR) within 3 months after LT was successfully associated with an annual decrease of eGFR (r(2) value = 0.574, P < .0001). CONCLUSION: Renal dysfunction is the most frequent chronic complication after LT. As chronic individual eGFR can be now accurately predicted with deterioration speed, recipient strata for renal protection strategies should be precisely targeted.

Plasma and synovial fluid levels of granulocytal elastase-alpha-1-protease inhibitor complex in patients with rheumatoid arthritis.

Plasma and synovial fluid levels and granulocytal elastase-alpha-1-protease inhibitor complex (EIC) in patients with rheumatoid arthritis (RA) were measured by enzyme-linked immunoassay and the results compared with those in patients with osteoarthrosis (OA). It was found that the plasma and synovial fluid levels of EIC in RA patients were higher than those in OA patients. There was a positive correlation between plasma EIC level in RA patients in Lansbury's index score of disease activity, as this tends to be higher when titer of RAHA in the plasma is high. The level of EIC in the synovial fluids correlated positively with granulocyte count and alpha-1-protease inhibitor (alpha-1-PI) level, and this, too, tends to be higher when titer of RAHA in synovial fluid is high. The results suggested that the level of EIC in the plasma or synovial fluids can be a good marker for the systemic or localized activation of the granulocytes and that IgM rheumatoid factor (IgMRF) is involved in the mechanism of the release of elastase.

Study on polymorphonucleocytal elastase-alpha 1-protease inhibitor complex in bronchoalveolar lavage fluid (BALF).

The purpose of the present study is to determine polymorphonucleocytal elastase-alpha 1-protease inhibitor complex in bronchoalveolar lavage fluid (BALF) in order to investigate the clinical significance of the increased complex in BALF. BALF was collected by the usual method using a bronchial fiberscope and concentrated 5-10 folds by Amicon UM-2 ultrafiltration. Determination of the complex in BALF and plasma was made in accordance with the method of Neumann et al. The level of complex (microgram/mg albumin, mean value) in BALF was 0.38 in healthy non-smokers (n = 5), 3.14 in healthy smokers (n = 6), 5.61 in the patients with chronic obstructive pulmonary disease (COPD) (n = 9) and 1.78 in the patients with idiopathic pulmonary fibrosis (IPF) (n = 15), that is, higher in healthy smokers and COPD and IPF patients. The ratio of the complex in BALF vs plasma was higher in healthy smokers and in COPD patients than in healthy non-smokers. Furthermore, there was a positive correlation between the level of this complex in BALF and the number of PMN in BLAF (r = 0.68, p less than 0.01). From the above it was assumed that the release of elastase from PMN is progressed in the lower airway of smokers and COPD patients.

Immunohistochemical quantitation for extracellular matrix proteins in rats with glomerulonephritis induced by monoclonal anti-Thy-1.1 antibody.

Extracellular matrix proteins (type I collagen and fibronectin) in frozen histologic sections of kidney cortex from rats with glomerulonephritis induced by a single intravenous administration of anti-Thy 1.1 antibody were quantified using an immunohistochemical micromethod. Type I collagen and fibronectin contents in renal cortex of rats with experimental glomerulonephritis (4.33 +/- 0.79 and 10.41 +/- 2.01 microgram/mg of total protein, respectively) were 262% and 151%, respectively, higher than in control rats given normal mouse IgG (1.65 +/- 0.16 and 6.88 +/- 0.95 microgram/mg, respectively; p < 0.01 in each case). In the glomerulonephritic rats, the increase in the contents of extracellular matrix proteins, especially type I collagen, correlated with increasing glomeruli with expansion of mesangial areas. The increase in type I collagen content correlated well with increasing urinary protein excretion and blood urea nitrogen and serum total cholesterol levels (r = 0.851, 0.812, and 0.837, respectively; p < 0.05 in each case). The decrease in creatinine clearance correlated with increasing content of type I collagen (r = 0.781; p < 0.05). The immunohistochemical micromethod may make it possible to evaluate the histopathological diagnosis of mesangial proliferative glomerulonephritis quantitatively.