Hemoadsorption of high-mobility group box chromosomal protein 1 using a column for large animals.

BACKGROUND: High-mobility group box chromosomal protein 1 (HMGB1) has recently been identified as an important mediator of various kinds of acute and chronic inflammation. A method for efficiently removing HMGB1 from the systemic circulation could be a promising therapy for HMGB1-mediated inflammatory diseases. MATERIALS AND METHODS: In this study, we produced a new adsorbent material by chemically treating polystyrene fiber. We first determined whether the adsorbent material efficiently adsorbed HMGB1 in vitro using a bovine HMGB1 solution and a plasma sample from a swine model of acute liver failure. We then constructed a column by embedding fabric sheets of the newly developed fibers into a cartridge and tested the ability of the column to reduce plasma HMGB1 levels during a 4-hour extracorporeal hemoperfusion in a swine model of acute liver failure. RESULTS: The in vitro adsorption test of the new fiber showed high performance for HMGB1 adsorption (96% adsorption in the bovine HMGB1 solution and 94% in the acute liver failure swine plasma, 2 h incubation at 37°C; p < 0.05 vs. incubation with no adsorbent). In the in vivo study, the ratio of the HMGB1 concentration at the outlet versus the inlet of the column was significantly lower in swine hemoperfused with the newly developed column (53 and 61% at the beginning and end of perfusion, respectively) than in those animals hemoperfused with the control column (94 and 93% at the beginning and end of perfusion, respectively; p < 0.05). Moreover, the normalized plasma level of HMGB1 was significantly lower during perfusion with the new column than with the control column (p < 0.05 at 1, 2, and 3 h after initiation of perfusion). CONCLUSION: These data suggest that the newly developed column has the potential to effectively adsorb HMGB1 during hemoperfusion in swine.

Increased plasma levels of high mobility group box 1 in patients with acute liver failure.

BACKGROUND: High-mobility group box 1 (HMGB1) is a monocyte-derived late-acting inflammatory mediator, which is released in conditions such as shock, tissue injury and endotoxin-induced lethality. In this study, we determined the plasma and hepatic tissue levels of HMGB1 in patients with acute liver failure (ALF). PATIENTS AND METHODS: We determined the plasma levels of HMGB1 and aspartate aminotransferase (AST) in 7 healthy volunteers (HVs), 40 patients with liver cirrhosis (LC), 37 patients with chronic hepatitis (CH), 18 patients with severe acute hepatitis (AH), and 14 patients with fulminant hepatitis (FH). The 14 patients with FH were divided into two subgroups depending upon the history of plasma exchange (PE) before their plasma sample collection. The hepatic levels of HMGB1 were measured in tissue samples from 3 patients with FH who underwent living-donor liver transplantation and from 3 healthy living donors. Hepatic tissue samples were also subjected to immunohistochemical examination for HMGB1. RESULTS: The plasma levels of HMGB1 (ng/ml) were higher in patients with liver diseases, especially in FH patients with no history of PE, than in HVs (0.3 ± 0.3 in HVs, 4.0 ± 2.0 in LC, 5.2 ± 2.6 in CH, 8.6 ± 4.8 in severe AH, 7.8 ± 2.7 in FH with a history of PE, and 12.5 ± 2.6 in FH with no history of PE, p < 0.05 in each comparison). There was a strong and statistically significant relationship between the mean plasma HMGB1 level and the logarithm of the mean AST level (R = 0.900, p < 0.05). The hepatic tissue levels of HMGB1 (ng/mg tissue protein) were lower in patients with FH than in healthy donors (539 ± 116 in FH vs. 874 ± 81 in healthy donors, p < 0.05). Immunohistochemical staining for HMGB1 was strong and clear in the nuclei of hepatocytes in liver sections from healthy donors, but little staining in either nuclei or cytoplasm was evident in specimens from patients with FH. CONCLUSION: We confirmed that plasma HMGB1 levels were increased in patients with ALF. Based on a comparison between HMGB1 contents in normal and ALF livers, it is very likely that HMGB1 is released from injured liver tissue.

Radiography using cosmic-ray electromagnetic showers and its application in hydrology.

In-situ measurements of soil water content provide important constraints on local/global hydrology. We demonstrate that the attenuation of the underground flux of cosmic-ray electromagnetic (EM) particles can be used to monitor the variation of soil water content after rainfalls. We developed a detection system that preferably selects EM particles by considering the coincidence of distant plastic scintillators. The calibration test beneath the water pool revealed that the count rate decreased by 0.6-0.7% with a 1 cm increase in the water level. The field measurement performed in the horizontal tunnel showed that the count rate dropped according to 48-h precipitation, after correcting the effects originating from atmospheric and water vapour pressures. These characteristics were confirmed using dedicated Monte Carlo simulations. This new method is called cosmic electromagnetic particle (CEMP) radiography.

Bedrock sculpting under an active alpine glacier revealed from cosmic-ray muon radiography.

Mountain glaciers form landscapes with U-shaped valleys, roche moutonées and overdeepenings through bedrock erosion. However, little evidence for active glacial carving has been provided particularly for areas above the Equilibrium Line Altitude (ELA) where glaciers originate. This is mainly due to our lack of information about the shape of the bedrock underneath active glaciers in highly elevated areas. In the past years, the bedrock morphology underneath active glaciers has been studied by geophysical methods in order to infer the subglacial mechanisms of bedrock erosion. However, these comprise surveys on the glaciers' surface, from where it has been difficult to investigate the lateral boundary between the ice and the bedrock with sufficient resolution. Here we perform a muon-radiographic inspection of the Eiger glacier (Switzerland, European Alps) with the aid of cosmic-ray muon attenuation. We find a reach (600 × 300 m) within the accumulation area where strong lateral glacial erosion has cut nearly vertically into the underlying bedrock. This suggests that the Eiger glacier has profoundly sculpted its bedrock in its accumulation area. This also reveals that the cosmic-ray muon radiography is an ideal technology to reconstruct the shape of the bedrock underneath an active glacier.

The vascular endothelial growth factor/fms-like tyrosine kinase system in human ovary during the menstrual cycle and early pregnancy.

In human ovaries, angiogenesis is known to be associated with the development of follicles and the formation of the corpus luteum (CL). A complex vascular network is formed within the thecal cell layer during follicular growth, and rapid neovascularization occurs toward the granulosa cell layer after ovulation. Vascular endothelial growth factor (VEGF) is a multifunctional cytokine, stimulating endothelial cell growth and enhancing microvascular permeability. A specific receptor for VEGF, fms-like tyrosine kinase (Flt-1), is expressed in vascular endothelial cells that mediates the action of VEGF. We examined the localization and expression of VEGF and Flt-1, using an immunohistochemical technique and RT-PCR analysis, in human follicles and corpora lutea during the normal menstrual cycle and early pregnancy. We measured concentrations of VEGF in extracts of human CL using an enzyme-linked immunosorbent assay during the luteal phase and early pregnancy. Immunostaining for VEGF was observed in granulosa cells from small antral follicles to preovulatory follicles. The staining was detected in thecal cells from medium-sized to preovulatory follicles. The intensity of the staining was gradually increased as a follicle grew. Flt-1 was localized in granulosa and thecal cells of preovulatory follicles as well as in endothelial cells. In the human CL, the intense staining for VEGF was observed in granulosa and thecal lutein cells, especially in the midluteal phase. The immunostaining for Flt-1 was faint in endothelial cells in the CL, whereas it was distinct in granulosa and thecal lutein cells. The concentrations of VEGF in lutein extracts were high in the early and midluteal phases and tended to decrease toward the late luteal phase. During early pregnancy, a measurable amount of VEGF was detected. RT-PCR analysis demonstrated that messenger ribonucleic acids encoding VEGF121, VEGF165, and Flt-1 were expressed in the CL. These results suggest that VEGF might have an autocrine role in the ovulatory process and luteal function as well as a paracrine role in angiogenesis.

Polymerase chain reaction-based microsatellite polymorphism analysis of follicular and Hürthle cell neoplasms of the thyroid.

Follicular and Hürthle cell carcinomas of the thyroid cannot be differentiated from adenomas by either preoperative fine needle aspiration or intraoperative frozen section examination, and yet there exist potentially significant differences in the recommended surgical management. We examined, by PCR-based microsatellite polymorphism analysis, DNA obtained from 83 thyroid neoplasms [22 follicular adenomas, 29 follicular carcinomas, 20 Hürthle cell adenomas (HA), and 12 Hürthle cell carcinomas (HC)] to determine whether a pattern of allelic alteration exists that could help distinguish benign from malignant lesions. Alterations were found in only 7.5% of informative PCR reactions from follicular neoplasms, whereas they were found in 23.3% of reactions from Hürthle cell neoplasms. Although there were no significant differences between follicular adenoma and follicular carcinoma, HC demonstrated a significantly greater percentage of allelic alteration than HA on chromosomal arms 1q (P < 0.001) and 2p (P < 0.05) by Fisher's exact test. The documentation of an alteration on either 1q or 2p was 100% sensitive and 65% specific in the detection of HC (P < 0.0005, by McNemar's test). In conclusion, PCR-based microsatellite polymorphism analysis may be a useful technique in distinguishing HC from HA. Potentially, the application of this technique to aspirated material may allow this distinction preoperatively and thus facilitate more optimal surgical management. Consistent regions of allelic alteration may also indicate the locations of critical genes, such as tumor suppressor genes or oncogenes, that are important in the progression from adenoma to carcinoma. Finally, this study demonstrates that Hürthle cell neoplasms, now considered variants of follicular neoplasms, differ significantly from follicular neoplasms on a molecular level.

Adrenal imaging with 131I-19-iodocholesterol in the diagnostic evaluation of patients with aldosteronism.

The results of adrenal scintiscans, venograms and venous aldosterone levels are compared with the histologic findings in 33 patients submitted to operations for primary aldosteronism. Standard and suppression scintiscans were performed 2-14 days following intravenous administration of 2mCi of 131I-19-iodocholesterol. The adrenal lesions were histologically classified into four categories: 25 patients had adenomas, 6 had macronodular hyperplasia, 1 had microscopic hyperplasia and 1 had an adenocarcinoma. Asymmetrical uptake between the two adrenals seen on standard scintiscans did not differentiate between a tumor or asymmetrical hyperplasia, unless the tumor was greater than 2 cm in diameter. During suppression scintiscans, unilateral uptake visible within five days of tracer injection was consistent with adenoma. Patients with nodular hyperplasia demonstrated early uptake in both adrenal glands during suppression scintiscans, while the patient with microscopic hyperplasia did not. The type of adrenal lesion was correctly identified in 20/26 (77%) of patients by suppression scintiscans; 21/28 (75% of patients by venograms and 12/16 (75%) of patients who had adrenal venous aldosterone measurements attempted. The majority of surgically correctible lesions could be identified on suppression adrenal scintiscans. Adrenal vein catheterization can be reserved for those patients in whom the results of suppression scintiscans are inconsistent with the clinical degree of aldosteronism.

Comparison of expressed sequence tags from male and female sexual organs of Marchantia polymorpha.

A total of 935 expressed sequence tags (ESTs) from male immature sexual organ were determined, of which 600 ESTs were assembled into 110 non-redundant groups, resulting in 445 unique EST sequences. Of these, 244 sequences shared significant similarities to known nucleotide or amino acid sequences in other organisms. The remaining 201 unique sequences showed no significant matches and thus are likely to be novel transcripts. ESTs from male and female immature sexual organs of a liverwort, Marchantia polymorpha, were compared to characterize gene expression patterns during sex differentiation. Ninety-nine male ESTs turned out to be common genes found also in the female library. Interestingly, one of the ESTs found only in male shows a significant similarity to the transformer-2 gene involved in sex determination in Drosophila. In female, several unique lectin ESTs were found that are not present in the male library.

Platelet-activating factor in hepatic ischemia/reperfusion injury. The effects of a PAF antagonist combined with a prostaglandin I2 analogue.

The effects of TCV-309, a specific platelet-activating factor (PAF) antagonist, and OP-41483, a prostaglandin I2 analogue, on warm ischemia/reperfusion injury of the rat liver were studied. Rats were divided into five groups by the duration of warm ischemia and the treatment used. The NS1 group (normal saline pretreatment) had 60 min of warm ischemia, while the NS2 group (normal saline pretreatment), the PGI2 group (OP-41483, 500 ng/kg/min pretreatment), the TCV group (TCV-309, 3 micrograms/kg), and the PGI2+TCV group (both the above dosages) underwent 120 min of warm ischemia. Postoperative survival after 30 days, bile secretion, serum endotoxin levels, and tissue glutathione levels after 60 min of reperfusion were compared between the groups. The survival rates for the NS1, NS2, PGI2, TCV, and PGI2+TCV groups were 80%, 0%, 50%, 80%, and 86.7%, respectively. Bile secretion, which has a strong correlationship with hepatic cellular ATP level, was strongly correlated with survival. The NS2 group had a high serum endotoxin level--however, the PGI2 and PGI2+TCV groups had normal levels. Although there were some discrepancies between survival and the tissue glutathione level, combined treatment with the PGI2 analogue and TCV-309 was most effective in inhibited oxidative stress. In conclusion, TCV-309 increased the survival rate after 120 min of warm hepatic ischemia without endotoxemia by the PGI2 analogue. This finding suggest that warm ischemia/reperfusion injury is related to the generation of PAF. Combined pretreatment with TCV-309 and a PGI2 analogue may be useful in liver transplantation.

The role of endothelin in the pathophysiology of renal impairment during acute liver rejection.

We assessed the role of endothelin in the development of renal dysfunction during acute rejection by examining the effect of a selective endothelin A (ETA) receptor antagonist BQ-123 in rats with acute liver rejection. Serum endothelin levels and endogenous creatinine clearance (Ccr) were monitored on days 1, 3, 5, 7, and 9 postoperatively. As indicators of renal hemodynamics, the estimated hemoglobin concentration of renal tissue (IHb) and the oxygen saturation of hemoglobin in renal blood (ISO2) were determined by reflectance spectrophotometry. In addition, the clearance of inulin and P-aminohippurate were determined, and the renal tissue blood flow was estimated by laser-Doppler flowmetry (LDF). As a model of allograft rejection, Lewis rats were transplanted orthotopically with DA rat livers. The serum endothelin level of allografted rejectors was significantly (P < 0.05) higher than that of isografted controls (Lewis rats with Lewis livers) on postoperative day 5, and it increased to a maximum of 5.38 +/- 0.95 pg/ml on day 9 (versus 1.23 +/- 0.18 pg/ml preoperatively). The values of Ccr, IHb, and ISO2 were all significantly (P < 0.05) lower in allografted rejectors than in isografted controls on day 5, and subsequently declined to a minimum on day 9 (P < 0.01). Treatment of allografted rejectors with BQ-123 markedly improved the renal parameters to levels similar to those in the isografted controls. These results strongly suggest that endogenous endothelin may play an important role in the development of renal impairment during acute liver rejection by reducing renal blood flow through binding with ETA receptor.

Hepatic release of endothelin-1 after warm ischemia. Reperfusion injury and its hemodynamic effect.

This study investigated the release of endothelin (ET)-1 from the liver after warm ischemia/reperfusion (I/R) injury. Wistar rats were subjected to 120 min of warm hepatic ischemia by clamping the hepatic hilum under porto-jugular shunting. Reperfusion was performed by unclamping. The rats were divided into 2 groups receiving intravenous treatment with an anti-ET-1 mAb before ischemia (AET group) and with mouse immunoglobulin G (sham group). Hepatic blood flow was assessed by laser-Doppler flowmetry and reflectance spectrophotometry and was compared between the 2 groups along with the bile flow rate. The ET-1 concentrations of hepatic venous and portal blood were determined in the sham group, and the portal blood endotoxin levels were assayed in both groups. Both groups developed transient hypotension after reperfusion, but hepatic blood flow subsequently showed a significant improvement in the AET group. Hepatic congestion was detected in the sham group by both reflectance spectrophotometry and histological examination. After reperfusion, bile flow was significantly greater in the AET group. The portal endotoxin concentration showed no increase in both groups, and the hepatic venous blood ET-1 level in the sham group was significantly higher until 3 hr after reperfusion compared to the portal blood level. The 30-day survival rate was 50% in the AET group, whereas all the sham rats died within 12 hr. ET-1 was released from the liver after I/R injury and apparently participated in systemic and local hemodynamic changes that affected survival.

Survival time and "cure" in papillary and follicular thyroid carcinoma with distant metastases: statistics following University of Michigan therapy.

Between 1947 and 1980, 103 patients with well-differentiated thyroid carcinoma with metastases outside the neck were treated with sodium iodide (I-131) after surgical treatment. Forty-one patients had distant metastases first detected an average of 7.44 yr after the initial operation establishing the diagnosis of thyroid carcinoma. Follicular and papillary carcinomas gave the same survival time in patients, matched for age and sex, who had metastases outside the neck. Those considered to be free of their metastatic disease after I-131 therapy survived three times as long as those with persistent disease. Patients freed of their metastases had a higher conformity rate with half of our ten procedures of "ideal" treatment, compared with patients not freed of their metastases.

Effect of large doses of 131I-19-iodocholesterol on metapyralone-induced adrenal cortical hyperplasia in dogs.

The potential use of 131I-19-iodocholesterol to treat ACTH excess Cushing's disease was evaluated in the dog. Three normal female dogs were given LD50 radiation doses of 131I-19-iodocholesterol without producing gross or histopathologically demonstrable change of the adrenals at autopsy 3 months later. The adrenal cortices of three dogs were made hyperplastic (to simulate the adrenal cortex in Cushing's disease) with ACTH and three with Metapyralone. In addition these six dogs were given KD50 doses of 131I-19-iodocholesterol. Three months after treatment, the adrenal glands of the ACTH-treated dogs were not enlarged, the cortex was thicker than normal, and there were no changes attributable to irradiation. At 3 months, the Metapyralone-treated dogs had enlarged adrenals, widening of the adrenal cortex, and no necrosis or other changes attributable to irradiation. It is concluded that a therapeutic trial of 131I-19-iodocholesterol in the treatment of Cushing's disease is not indicated.

The morphologic effects of simultaneous infusion of levarterenol and phentolamine on the canine myocardium.

The effect of simultaneous administration of levarterenol and phentolamine on the myocardium over a 3 hour period was studied in 15 adult mongrel dogs. All animals receiving levarterenol alone had moderate-to-severe subendocardial hemorrhage and necrosis. Four of the 6 animals receiving the simultaneous infusion of levarterenol and phentolamine had little or no hemorrhage or necrosis. These differences are significant (p less than 0.02). It is concluded that the administration of phentolamine simultaneously with levarterenol affords a significant protective effect on the myocardium.

Parathyroid hamartoma: report of two cases and review of the literature.

Parathyroid hamartomas are rare. Only four cases, two of which are associated with clinical hyperparathyroidism, have been reported in the literature. This is a report of two additional cases of functioning parathyroid hamartomas, one accompanied by proliferating fat and the other by a myxomatous, fibrillar stroma.

The identification of lupus erythematosus cells. A comparison of supravital fluorochromation with a standard Wright's stain method.

A comparison of standard Wright's stained lupus erythematosus preparations and an acridine orange fluorochromatic method was conducted using 354 consecutive lupus erythematosus preparations involving 264 patients. The results of this comparison and a discussion of the fluorochromatic procedure are presented.

Effects of several commonly used fixatives on DNA and total nuclear protein analysis by flow cytometry.

Five commonly used fixatives (AZF, B-5, Bouin's, formalin, and Zenker's) were evaluated for their effect on the flow cytometric analysis of DNA and total nuclear protein (TNP) in solid tumors. Data were obtained with the use of colonic adenocarcinoma, squamous carcinoma of the lung, mammary adenocarcinoma, and spleen with a plasma cell leukemic infiltrate. The parameters examined were G0-G1 DNA staining intensity, %G0-G1, percent coefficient of variation (%CV), percent debris, and TNP staining intensity. The results showed that variations in the fixation of solid tumor significantly affected flow cytometric-derived parameters. In this study, paraffin-embedded tissue (PET) fixed in 10% (v/v) neutral buffered formalin (NBF) produced the best results, with a %CV below 4.7, whereas fixatives such as Zenker's and B-5 produced poor %CVs (above 6.0) or uninterpretable TNP and light scatter data. These data suggest that a portion of all tissue samples be fixed in NBF to allow for subsequent analysis by fixative-sensitive assays such as DNA in situ hybridization and flow cytometry.

Percutaneous tetracycline instillation for sclerosis of recurrent thyroid cysts.

Of 770 hypofunctioning thyroid nodules subjected to needle biopsy since 1977, the fluid of 172 cysts (22%) was aspirated. The fluid, cytologically examined, showed no evidence of carcinoma except in three instances: papillary carcinomas that were subsequently treated surgically. Of the cystic lesions, 19 (11%) recurred and were treated by reaspiration and the instillation of tetracycline hydrochloride into the cyst cavity. In all but one instance this resulted in obliteration of the cysts (95%). This is a considerably higher rate of success than that reported from aspiration alone (40% to 85%). The follow-up period ranged from 1 to 42 months, with no recurrence or subsequent development of a thyroid nodule in these patients. The one recurrent cyst was reaspirated and reinstilled a second time, which resulted in great reduction in size. Not considered for this procedure were patients in whom the cytologic condition of the fluid was abnormal or indicative of a malignancy. Also excluded were those in whom there was incomplete decompression of the cyst, manifested by a persistent nodule. No patients had a history of head or neck irradiation or cervical lymphadenopathy, both contraindications for this procedure. No patients had grossly bloody aspirates. Recurrence of thyroid cysts after aspiration was not thought to be, in itself, a criterion for surgical selection. The effective use of tetracycline hydrochloride as a sclerosing agent in the treatment of malignant pleural effusions is well documented. Its action in producing symphysis is thought to be related to its low pH (2.0). This procedure is safe, simple, cost effective, and well tolerated by patients. It obviates the need for excision in patients who fulfill the aforementioned criteria.

Overexpression of p53 in tall cell variants of papillary thyroid carcinoma.

BACKGROUND: The tall cell variant (TCV) is a clinically aggressive subtype of papillary thyroid cancer. The aim of this study was to discover the prevalence of mutant forms of p53 protein in this subtype and relate it to clinical outcome. METHODS: Eighteen patients with TCV and a control group with common papillary cancers, matched for age and gender, were studied. The p53 mutations were identified by means of immunohistochemical staining. Data reviewed were overall survival, recurrence, TNM stage, and p53 positivity. RESULTS: p53 mutations occurred in 11 (61%) patients with TCV compared with two (11%) in control group (p = 0.05). In the TCV group two patients died of the disease (11%) and eight (44%) had local recurrences or distant metastases compared with none in the control group. All deaths and 70% of the recurrences occurred in patients with stage III or IV disease. p53 positivity did not correlate with any reduction in survival (7% compared with 9%) but with increased rate of local (23% compared with 4%) and distant (23% compared with 13%) recurrences. CONCLUSIONS: TCV was associated with a significantly higher rate of p53 positivity than common papillary carcinoma. The stage of the disease seemed to be a better prognostic indicator than p53 positivity for overall survival.

Secretory diarrhea with islet cell hyperplasia and increased immunohistochemical reactivity to serotonin.

Two patients with secretory diarrhea and signs and symptoms consistent with the Verner-Morrison syndrome and islet cell hyperplasia are described. Both patients responded well to subtotal pancreatectomies. The morphologic changes in the pancreata were characterized by proliferation of islets associated with periductal and interstitial fibrosis. Immunohistochemical stains demonstrated increased staining for serotonin in islet cells. A few islet cells also stained for vasoactive intestinal polypeptide. The significance of these results is discussed.