A probe generated by chromosome microdissection, useful for analyzing Y chromosome evolution in Old World monkeys

We isolated a DNA probe, designated MMDYZ1, using a chromosome microdissection technique from the Y chromosome of the Rhesus monkey. The probe obtained from eight whole Y chromosomes shows higher specificity for the Y short arm of the Rhesus monkey, which consists totally of constitutive heterochromatin. Two microclones (MMY#3 and MMY#4) were constructed from the Y-specific primary PCR products. Sequence analysis of these two microclones revealed that both were essentially identical to each other and the sizes were 870 and 686bp, respectively. From alignment analysis using the Genbank database of primates, the alphoid DNA has the highest affinity with the probe. However, the total composition of this probe has extremely high homology with the Y short arm of the Rhesus monkey, as demonstrated by fluorescence in-situ hybridization (FISH). Comparative FISH-mapping disclosed that this DNA-sequence cluster was located at extremely different sites on the Y chromosome in several species of the Old World monkey. Accordingly, this probe seems to be a high-quality tool, now established for the first time, for investigating Y chromosome evolution of the Old World monkey.     

Hepatotoxicity caused by both tacrolimus and cyclosporine after living donor liver transplantation.

We present a case report of a posttransplant patient who had hepatotoxicity due to both tacrolimus and cyclosporine and cholestatic jaundice due to tacrolimus. The patient did not show sustained improvement in enzyme and bilirubin abnormalities after an initial change from tacrolimus to cyclosporine or with a change back to tacrolimus, but he ultimately showed improvement when the blood concentration of tacrolimus was lowered. A 56-year-old man with subacute fulminant hepatitis induced by acarbose was admitted to our hospital for living donor liver transplantation. The liver graft consisted of the left lobe from his ABO-identical son. The early posttransplant course was uneventful. The serum levels of aspartate aminotransferase (AST), alanine aminotransferase (ALT), and total bilirubin improved initially, but the ALT and AST levels later increased. A liver biopsy suggested a presumptive diagnosis of drug reaction. All drugs were discontinued, the immunosuppressive agent was changed from tacrolimus to cyclosporine. After initial improvement, the ALT and AST levels increased again. Assuming a reaction to cyclosporine, we decreased the concentration of cyclosporine in the blood. The enzyme levels improved temporarily but again began to rise. We changed the immunosuppressive agent to tacrolimus, which resulted in improvements in the ALT and AST levels; however, the total bilirubin level increased. We interpreted this increase as tacrolimus-induced cholestasis; in response, we decreased the blood concentration of tacrolimus to between 3 and 5 ng/dL and added 1,000 mg of mycophenolate mofetil to the drug regimen. The patient recovered without further complications. Repeated liver biopsies throughout the hospital course suggested that the mild mononuclear cell infiltration observed in a few triads had not been caused by acute rejection but had possibly been drug-induced.     

Genetically linked C-type lectin-related ligands for the NKRP1 family of natural killer cell receptors

The natural killer (NK) gene complex (NKC) encodes orphan lectin-like NK cell receptors that may explain uncharacterized NK cell specificities. Unlike other NKC-encoded receptors that recognize molecules with major histocompatibility complex (MHC) class I folds, here we show that mouse Nkrp1d and Nkrp1f bind specific C-type lectin-related (Clr) molecules. Nkrp1d mediated inhibition when recognizing Clrb, a molecule expressed in dendritic cells and macrophages. Nkrp1 (official gene name, Klrb1) and Clr are intertwined in a genetically conserved NKC region showing recombination suppression, reminiscent of plant self-incompatibility loci. Thus, these findings broaden the 'missing-self' hypothesis from solely involving MHC class I to including related NK cell receptors for lectin-like ligands, and reflect genetic strategies for biological self-recognition processes in other species.     

Isolation and characterization of two bladder carcinoma-associated antigens

By the use of mouse monoclonal antibodies we have earlier defined five distinct antigens associated with transitional cell carcinoma of the human urinary bladder (TCC). Two of these antigens have now been purified and partially characterized. For their purification from isolated tumor cell membranes, a rapid and efficient method of affinity chromatography was developed in which the coupling of monoclonal antibodies to protein A-Sepharose was fortified by treatment with glutaraldehyde. From these columns, 60-80% of the antigenic activity present in membrane lysates could be recovered by acid elution, corresponding to approximately 0.25-0.5 micrograms antigen protein per mg membrane protein added. The two isolated antigens are relatively hydrophobic membrane components not found in normal bladder tissue. The antigen defined by the monoclonal antibody S2C6 is confined to bladder carcinoma and B cells. It is a glycosylated, ConA-binding polypeptide with Mr 50 000 as established by SDS-PAGE. It is acidic with an IP of 3.2, heat stable up to 85 degrees C, stable at low pH (2.0) but sensitive to SDS. The antigen defined by the monoclonal antibody 7E9 is confined to bladder carcinoma but is also weakly expressed in some blood vessel endothelium. It consists of two main polypeptides of Mr 29,000 and 23,000 which do not bind to ConA. It has an IP of 7.4 and its antigenic activity is abolished by heating to 50 degrees C for 5 min. As the S2C6 antigen, it is stable at low pH but susceptible to SDS.     

Eversion and ligation of a diverticulum: report of an inspirational case and subsequent animal study

Although most patients with colonic diverticula have no symptoms, perforation or bleeding can be troublesome. Recently, we treated an elderly patient with a diverticular perforation of the sigmoid colon who required a loop colostomy and later resection of the diseased colon. Between the operations, the colostomy resembled a sea slug with horns, which represented eversions of the diverticula. This interesting phenomenon suggests that diverticula may be everted as pseudopolyps and ligated endoscopically. Therefore, the procedure was tested on a Landrace pig, of which the large bowel wall was pulled with an endoscope at 9 different sites using forceps (n=6) or suction (n=3) and ligated. Four days later, reexamination revealed total necrosis at 8 sites and ulceration at 1. The animal was well. The bowel removed 2 weeks later showed mucosal fold convergences without perforations. Our procedure can be used for prophylactic or therapeutic treatment of patients with colonic diverticula.     

Surgical treatment for better prognosis of patients with liver metastases from colorectal cancer

BACKGROUND/AIMS: Hepatic resection is established as the best available treatment for colorectal cancer metastases to the liver, but the optimal procedure for the resection is now controversial. In this study we retrospectively evaluated the influence of the selection of surgical procedures for hepatic resection in determining the prognostic factors. METHODOLOGY: A population of 111 patients with liver metastases from colorectal cancer underwent initial and radical hepatic resection. Survival rates as a function of clinical, primary tumor pathologic TNM classification and surgical determinates were examined retrospectively with univariate and multivariate analyses. RESULTS: The overall, 1-, 3-, 5- and 10-year survival rates were 85.5%, 51.4%, 41.6% and 30.9%, respectively. Patient survival was uncorrelated with any of the following factors: sex, age, temporal relationship, primary tumor site, tumor stage, pathological depth, pathological vascular invasion, pathological lymphatic invasion and pathological lymph node metastases of the primary tumor, maximum diameter of liver lesions, intrahepatic distribution patterns, type of hepatectomy, and number of hepatectomies. However, the number of liver tumors and the tumor-free margins were significantly associated with good prognosis. CONCLUSIONS: Safe hepatectomy with adequate tumor-free margins leads to a better prognosis.     

A morphometric study of subcortical changes in Alzheimer's disease

The populations and loss of neurons, the density of neurofibrillary tangles (NFT) and senile plaque (SP), and the macroscopic shrinkage of the subcortical areas in patients with Alzheimer's disease (AD) were quantitatively evaluated using computer-based image analysis techniques. Twelve subcortical areas were examined at distinct levels of the brain in eight AD cases and five age-matched controls. The areas examined were as follows: the nucleus basalis of Meynert (nbM); globus pallidus (GP); putamen (PT); claustrum (CL); the anterior thalamic nucleus (THA: anterior ventral thalamic nucleus); medial thalamic nucleus (THM: medio-dorsal thalamic nucleus); lateral thalamic nucleus (THL: ventro-lateral and ventro-anterior thalamic nuclei); subthalamic nucleus (ST); substantia nigra, zona compacta (SNC); red nucleus (RD); locus ceruleus (LC); and raphe nucleus (RP). Distinct neuronal loss and a high density of NFT were shown in the nbM, LC, RP, CL, SNC, and THA in the AD brain. Because of the high density of the NFT relative to the neuronal loss, neuronal loss in those nuclei may have been mainly caused by NFT formation. In the THM and THL, no distinct neuronal loss but some degree of NFT formations was detected. In the GP, ST, and RD, neuronal shrinkage or loss was shown in patients with AD. With respect to the SP, the density of SP in the CL, PT and THM was high (1.7–2.2%), whereas that in the SNC, GP, ST and RD was low. This finding suggests that SP may not have major effects on the neuronal degeneration observed in subcortical nuclei of patients with AD. Although macroscopic shrinkage of all subcortical nuclei was seen in AD patients, selective vulnerability to atrophic change was not evident at macroscopic levels. This comprehensive morphometric study offers more precise information for evaluating the subcortical dysfunction in AD patients than do conven-tional neuropathological examinations.     

Long-term results of multiple minocycline hydrochloride injections for the treatment of symptomatic solitary hepatic cyst

BACKGROUND: The current study presents the long-term results of multiple minocycline hydrochloride (MINO) injections for the treatment of symptomatic solitary hepatic cyst. METHODS: From 1989 to 1998, nine patients were referred for the treatment of a symptomatic solitary hepatic cyst, comprising two males and seven females aged between 41-72 years (mean 58.2 years). The chief complaint in all cases was upper abdominal discomfort or pain. Mean cyst diameter was 14.1 +/- 2.3 cm. After insertion of an 8 or 9-French catheter into the cyst, 200 mg of MINO in 10 mL saline was injected and the catheter was flushed with 10 mL saline, for a total of 20 mL saline. The catheter was then clamped for 30 min. MINO injection was performed daily for 7-8 days, after which time the catheter was removed. RESULTS: One patient complained of moderate right subscapular pain immediately after the injection, and one patient complained of pain at the site of catheter insertion. Patients were followed for 42-153 months (mean 85 months). Cyst regression without recurrence was documented in all patients. CONCLUSIONS: Multiple injections of MINO are safe, definitive treatments for symptomatic solitary hepatic cyst.     

Long-term results of elective hepatectomy for the treatment of ruptured hepatocellular carcinoma

Background/Purpose

According to the General rules for the clinical and pathological study of primary liver cancer, compiled by the Liver Cancer Study Group of Japan, ruptured hepatocellular carcinoma (HCC) is classified as stage 4, even if the tumor is small and solitary. We examined the long-term results of elective hepatectomy for the treatment of ruptured HCC.

Methods

A first hepatectomy was performed without operative death in 193 patients with HCC. Ten patients had ruptured HCC (ruptured group) and 183 patients had nonruptured HCC (nonruptured group). The extension of HCC was macroscopically classified as stage 1 in 23 patients, stage 2 in 71, stage 3 in 53, and stage 4 in 46.

Results

Cumulative survival rates in the ruptured group at 1, 5, and 10 years were 90.0%, 67.5%, and 20.3%, respectively. The cumulative survival rate was lower in patients with stage 4 disease in the nonruptured group than that in patients in the ruptured group (P < 0.05). Cumulative survival rates did not differ significantly between patients in the ruptured group and those with stage 2 or stage 3 disease.

Conclusions

Survival rates after elective hepatectomy in patients with ruptured HCC are good, even if the disease is classified as stage 4.