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Treatment intensification has improved survival in patients with hepatoblastoma (HB); however, these treatments are associated with an increased risk of late effects, including second malignant neoplasms (SMNs). Data is limited regarding SMNs following HB treatment. Cases of SMNs following treatment for HB reported in the literature and from personal communication were analyzed to further assess this late effect. Thirty-eight patients were identified. The median age at diagnosis of HB was 16 months (range: 3 to 168 mo). All patients had received a platinum agent, and almost all had anthracycline exposure. The SMNs reported were hematopoietic malignancies (n=19), solid tumors (n=12), and post-transplant lymphoproliferative disorder (n=7). Of the 36 patients with outcome data, 19 survived. SMNs following HB treatment were primarily seen in patients with chemotherapy exposure, a history of liver transplantation, hereditary tumor predisposition syndromes, and/or a history of radiation treatment. Hematopoietic malignancies were the most common SMN reported in this cohort and were diagnosed earlier than other SMNs. Prospective collection of data through a companion late effects study or international registry could be used to further evaluate the rates and risks of SMNs as well as tumor predisposition syndromes in patients treated for HB.

Background. Telomerase is an enzyme that adds hexameric TTAGGG nucleotide repeats onto the ends of vertebrate chromosomal DNAs (i.e., telomeres) to compensate for losses that occur with each round of DNA replication. Somatic cells do not have telomerase activity and stop dividing when the telomeric ends of at least some chromosomes have been shortened to a critical length. It has been suggested that immortalized cells (including some, but probably not all, cancer cells) continue to proliferate indefinitely because they express telomerase. Purpose: To investigate whether expression of telomerase is a prerequisite for the development of naturally occurring human cancers, we assayed the levels of telomerase activity in specimens of human lung tumor and adjacent normal tissue. Methods: Using a polymerase chain reaction-based assay, we examined telomerase activity in 136 primary lung cancer tissues and 68 adjacent noncancerous tissues obtained by surgical resection. We also studied telomerase activity in four primary and 23 metastatic lesions obtained through biopsy (two patients) or autopsy (10 patients). Relative telomerase activity levels were estimated by serial dilutions of extracts prepared from the specimens. Telomerase activity was also assayed in extracts of cells present in pleural fluids from three patients with adenocarcinoma of the lung. Results: Among surgically resected samples, telomerase activity was detected in 109 (80.1%) of 136 primary lung cancer tissues and in three (4.4%) of 68 normal adjacent tissues. All 11 surgically resected specimens of primary small-cell lung cancer (from 11 patients) revealed high levels of telomerase activity, whereas the activity ranged from undetectable to high levels in the 125 surgically resected specimens of primary non-small-cell lung cancer tissue (from 125 patients). Generally, high levels of telomerase activity were observed in metastatic lesions and tumors with altered telomere length. A few primary and, surprisingly, some metastatic tumors did not appear to have detectable telomerase activity. Telomerase activity was, however, detected in cells present in all tested pleural fluids obtained (from three patients with adenocarcinoma of the lung). Conclusion: The subset of non-small-cell lung cancers that exhibits only low or undetectable levels of telomerase activity may contain primarily mortal cancer cells. Cancers that exhibit high levels of telomerase activity, such as all of the small-cell lung cancers examined in this study, are likely to consist mainly of immortal cells. Implications: Telomerase activity may be useful both as a diagnostic marker to detect the existence of immortal lung cancer cells in clinical materials and as a target for therapeutic intervention. [J Natl Cancer Inst 87:895-902, 1995]

Background. Most patients with neuroblastoma who present with opsomyoclonus have a good prognosis. Neuroblastomas from such patients have been reported to contain a single copy of the N-myc gene. The authors describe three cases of patients with advanced neuroblastoma with opsomyoclonus, which had poor outcomes despite multimodal therapy. Methods. Amplification and expression of the N-myc gene were examined in these three primary tumors using Southern and Northern blot analyses. Then, flow cytometric analysis of the cellular DNA contents of these tumors was performed. Results. N-myc amplification was observed in two tumors and N-myc RNA overexpression was observed in all three. Analysis of the cellular DNA contents of the tissue specimens revealed hyperdiploidy in all three tumors; one had a triploid index and the other two had hypotetraploid indexes. Conclusions. These findings suggest that a poor outcome for patients with opsomyoclonus may be associated with N-myc gene activation (amplification and/or overexpression) and that a hyperdiploid tumor is not always associated with a good prognosis. Cancer 1994; 74:1821-6.

The most recent advance in the care of children diagnosed with hepatoblastoma and hepatocellular carcinoma is the Pediatric Hepatic International Tumor Trial, which opened to international enrollment in 2018. It is being conducted as a collaborative effort by the pediatric multicenter trial groups in North America, Europe, and the Far East. This international effort was catalyzed by a new unified global risk stratification system for hepatoblastoma, an international histopathologic consensus classification for pediatric liver tumors, and a revised 2017 collaborative update of the PRE-Treatment EXTent of disease radiographic based staging system.

Background: Some neuroblastoma (NB) cases are suitable for minimally invasive surgery (MIS), but indication and technical issue are unclear. We assessed the current status of MIS for abdominal NB after mass screening period in Japan. Methods: Preliminary questionnaires requesting the numbers of NB cases that underwent MIS from 2004 to 2016 were sent to 159 Japanese institutes of pediatric surgery. The secondary questionnaires were then sent to the institutions that reported MIS cases of NB in order to collect detailed data. Results: One hundred and thirty-four (84.2%) institutions responded to the preliminary questionnaires, and 83 (52.2%) reported managing operative cases. The total number of operative cases was 1496. MIS was performed for 175 (11.6%) cases, of which the completed forms of 140 patients were returned, including 100 abdominal NB cases. The male/female ratio was 51/49. Forty-seven cases underwent a laparoscopic biopsy, and 2 (4.3%) cases were converted to laparotomy due to bleeding. Sixty-five cases underwent MIS for radical resection, and 7 (10.8%) were converted to laparotomy. The reasons for open conversion were bleeding and severe adhesion. Regarding open conversion, there were no significant relationships between conversion and neo-adjuvant chemotherapy, biopsies, stage, size, or MYCNamplification. We found no relationship between resectability and vascular encasement in this study. There was relationship between the resected tumor size and the patients’ height, which was expressed using the following formula: $y=0.0316x+1.4812$\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$y = 0.0316x + 1.4812$$\end{document}(x, patients height, y, tumor size; p= 0.004219, SE: 1.55566). Postoperative complications after radical resection were recognized in 7 (10.8%) cases. Conclusions: MIS was performed in limited cases of abdominal NB. A laparoscopic biopsy with careful attention to bleeding is feasible. The resected tumor size was shown to correlate with the patients’ height. Tumor size within 6 cm of maximum diameter can be resected safely.

Monitoring drug uptake, its metabolism, and response on the single-cell level is invaluable for sustaining drug discovery efforts. In this study, we show the possibility of accessing the information about the aforementioned processes at the single-cell level by monitoring the anticancer drug tamoxifen using live single-cell mass spectrometry (LSC–MS) and Raman spectroscopy. First, we explored whether Raman spectroscopy could be used as a label-free and nondestructive screening technique to identify and predict the drug response at the single-cell level. Then, a subset of the screened cells was isolated and analyzed by LSC–MS to measure tamoxifen and its metabolite, 4-Hydroxytamoxifen (4-OHT) in a highly selective, sensitive, and semiquantitative manner. Our results show the Raman spectral signature changed in response to tamoxifen treatment which allowed us to identify and predict the drug response. Tamoxifen and 4-OHT abundances quantified by LSC–MS suggested some heterogeneity among single-cells. A similar phenomenon was observed in the ratio of metabolized to unmetabolized tamoxifen across single-cells. Moreover, a correlation was found between tamoxifen and its metabolite, suggesting that the drug was up taken and metabolized by the cell. Finally, we found some potential correlations between Raman spectral intensities and tamoxifen abundance, or its metabolism, suggesting a possible relationship between the two signals. This study demonstrates for the first time the potential of using Raman spectroscopy and LSC–MS to investigate pharmacokinetics at the single-cell level.

Herein, we report two paediatric cases of ovarian teratoma-associated anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis. We performed surgical resection of the teratomas—enucleation in case 1 and salpingo-oophorectomy in case 2. The pathological diagnoses were immature teratomas with a small number of neuroglial elements, and cerebrospinal fluid (CSF) anti-NMDAR antibody test was positive in both cases. In case 1, the patient showed excellent recovery, and in case 2, neurological symptoms gradually improved. Paediatric cases of ovarian teratoma-associated anti-NMDAR encephalitis are rare, and early surgical resection and immunotherapy are important for good prognosis. We highlight the occurrence of paediatric cases of ovarian teratoma-associated anti-NMDAR encephalitis to create awareness and highlight the importance of surgical resection despite severe neurological symptoms.

Liver tumors are rare in children, and their diagnoses may be challenging particularly because of the lack of a current consensus classification system. Systematic central histopathological review of these tumors performed as part of the pediatric collaborative therapeutic protocols has allowed the identification of histologic subtypes with distinct clinical associations. As a result, histopathology has been incorporated within the Children’s Oncology Group (COG) protocols, and only in the United States, as a risk-stratification parameter and for patient management. Therefore, the COG Liver Tumor Committee sponsored an International Pathology Symposium in March 2011 to discuss the histopathology and classification of pediatric liver tumors, and hepatoblastoma in particular, and work towards an International Pediatric Liver Tumors Consensus Classification that would be required for international collaborative projects. Twenty-two pathologists and experts in pediatric liver tumors, including those serving as central reviewers for the COG, European Société Internationale d’Oncologie Pédiatrique, Gesellschaft für Pädiatrische Onkologie und Hämatologie, and Japanese Study Group for Pediatric Liver Tumors protocols, as well as pediatric oncologists and surgeons specialized in this field, reviewed more than 50 pediatric liver tumor cases and discussed classic and newly reported entities, as well as criteria for their classification. This symposium represented the first collaborative step to develop a classification that may lead to a common treatment-stratification system incorporating tumor histopathology. A standardized, clinically meaningful classification will also be necessary to allow the integration of new biological parameters and to move towards clinical algorithms based on patient characteristics and tumor genetics, which should improve future patient management and outcome.

As a rare pediatric tumor, hepatoblastoma presents challenges to the individual practitioner as no center will see more than a handful of cases each year.

The aim of this study was to investigate the relationship between histological findings of the pancreatic stump and postoperative pancreatic exocrine function.

Pancreatic islet transplantation requires multiple transplants to achieve insulin independence. Only one third of the islet mass is stably engrafted; one of the causes of which is ascribed to oxidative stress. We confirmed the hypothesis that administration of edaravone, a free radical scavenger, in the early posttransplantation period promotes islet cell engraftment.

Human telomerase reverse transcriptase (hTERT), one of the subunits of telomerase, is a promising diagnostic marker for pancreatic cancer. In the present study, we did a large-scale analysis of 115 preoperative pancreatic juice specimens to evaluate the feasibility of detection of hTERT expression by immunohistochemistry for preoperative diagnosis of pancreatic malignancy.

Background: Epithelial membrane protein 3 (EMP3), was recently reported to be a tumor suppressor gene for several solid tumors, and is drawing attention as a novel prognostic marker, since its expression level or hypermethylation of the promoter region is associated with clinical prognosis in neuroblastoma and esophageal cancer. However, some controversial data were also observed in gliomas and breast cancers, and there seems to be more than deletion/hypermethylation to its silencing mechanisms. Objective: To clarify the discrepancies in the biological behavior of EMP3 among the different organ-derived malignancies or histologies and validate the potential of EMP3as a tumor suppressor for solid tumors. Methods: Literature dealing with EMP3in the PubMed database was reviewed. Results/conclusions: EMP3is a novel tumor suppressor gene in some kinds of malignancies, but not all, at the step of cellular immortalization rather than carcinogenesis. It may become a potent prognostic marker and a therapeutic target in such tumors.

We studied the characteristics of methicillin (meticillin)-resistant Staphylococcus aureus (MRSA) strains that caused enteritis. In a previous report, we demonstrated that both phenotypic and genotypic changes were associated with MRSA enteritis; and we hypothesized that the accessory gene regulator (agr), which is a global regulator of staphylococcal virulence and upregulates several exoproteins, is the key factor associated with the development of MRSA enteritis. In this study, we examined 12 MRSA isolates associated with enteritis from stool samples and 17 MRSA isolates not associated with enteritis that had the following characteristics: the strains associated with enteritis had the same genotype (genotype A), as detected by pulsed-field gel electrophoresis, or the strains were isolated from stools. The differences between strains that caused enteritis and those that did not cause enteritis strains were examined by quantitative reverse transcription-PCR to assess RNAII, agrA, RNAIII, and tst expression and by sequencing of the agr locus. The levels of expression of agrA, RNAIII, and tst were higher by the MRSA isolates associated with enteritis than by the MRSA isolates not associated with enteritis, whether or not they were of the same genotype. The levels of expression of RNAII by almost all the clinical isolates were similar. Sequencing of the agr locus showed that all MRSA isolates that caused enteritis have agr mutations, whereas the MRSA isolates that did not cause enteritis, with three exceptions, did not. Many of the isolates associated with enteritis had the same mutation, especially at the C-terminal end of agrA. These results suggest a trend in which mutations in the agr locus modify the expression of agrA and RNAIII and the production of toxin, all of which may increase the virulence and influence the occurrence of MRSA enteritis.

ABSTRACTWe studied the characteristics of methicillin (meticillin)-resistant Staphylococcus aureus(MRSA) strains that caused enteritis. In a previous report, we demonstrated that both phenotypic and genotypic changes were associated with MRSA enteritis; and we hypothesized that the accessory gene regulator (agr), which is a global regulator of staphylococcal virulence and upregulates several exoproteins, is the key factor associated with the development of MRSA enteritis. In this study, we examined 12 MRSA isolates associated with enteritis from stool samples and 17 MRSA isolates not associated with enteritis that had the following characteristics: the strains associated with enteritis had the same genotype (genotype A), as detected by pulsed-field gel electrophoresis, or the strains were isolated from stools. The differences between strains that caused enteritis and those that did not cause enteritis strains were examined by quantitative reverse transcription-PCR to assess RNAII, agrA, RNAIII, and tstexpression and by sequencing of the agrlocus. The levels of expression of agrA, RNAIII, and tstwere higher by the MRSA isolates associated with enteritis than by the MRSA isolates not associated with enteritis, whether or not they were of the same genotype. The levels of expression of RNAII by almost all the clinical isolates were similar. Sequencing of the agrlocus showed that all MRSA isolates that caused enteritis have agrmutations, whereas the MRSA isolates that did not cause enteritis, with three exceptions, did not. Many of the isolates associated with enteritis had the same mutation, especially at the C-terminal end of agrA. These results suggest a trend in which mutations in the agrlocus modify the expression of agrAand RNAIII and the production of toxin, all of which may increase the virulence and influence the occurrence of MRSA enteritis.

Abstract: Telomerase, a critical enzyme responsible ‘for cellular immortality, is usually repressed in somatic cells except for lymphocytes and self-renewal cells, but is activated in approximately 85% of human cancer tissues. The human telomerase reverse transcriptase (hTERT) is the catalytic component of human telomerase. In cancers in which telomerase activation occurs at the early stages of the disease, telomerase activity and hTERT expression are useful markers for the detection of cancer cells. In other cancers in which telomerase becomes upregulated upon tumor progression, they are useful as prognostic indicators. However, careful attention should be paid to false–negative results caused by the instability of telomerase and of the hTERT mRNA and the presence of PCR inhibitors, as well as to false–positive results caused by the presence of alternatively spliced hTERT mRNA and normal cells with telomerase activity. Recently, methods for the in situ detection of the hTERT mRNA and protein have been developed. These methods should facilitate the unequivocal detection of cancer cells, even in tissues containing a background of normal telomerase-positive cells.

Abstract: Hepatic partial ischemic/reperfusion (I/R) injury, in which ischemic and nonischemic areas of the liver are likely to respond to each other after reperfusion, often occurs following hepatobiliary surgical procedures. Kupffer cells (KCs) are considered to play a major role in hepatic I/R injury. To study the activation of KCs in ischemic and nonischemic liver tissues following hepatic I/R, we investigated the superoxide generation and proinflammatory cytokine production of KCs in both liver parts in a rat model of partial hepatic I/R injury. KC superoxide generation in the ischemic and nonischemic lobes was upregulated 6 and 24 h after reperfusion, respectively, and then accelerated. The production of interleukin-1β (IL-1β) by KCs in the ischemic lobes increased during the early and late phases, 6 h and 48–72 h after reperfusion, respectively. A late increase in IL-1β production was also observed in the nonischemic lobes. Production of tumor necrosis factor-α (TNF-α) increased 6–24 h after reperfusion in both lobes. Upregulation of IL-1β mRNA in the ischemic lobes preceded the upregulation of TNF-α mRNA in both lobes. The hepatic partial I/R process results in activation of KCs in ischemic and nonischemic areas of the liver. The KCs are activated during the early phase after reperfusion in the ischemic areas, followed by activation in both the ischemic and nonischemic areas. This could be a cause of liver dysfunction after partial hepatic I/R during surgery.

Neuroblastoma shows remarkable heterogeneity, resulting in favorable and unfavorable outcomes. It is well known that almost all cases with MYCN amplification have a poor prognosis. We have previously reported that unfavorable tumors show high telomerase activity, whereas favorable tumors show low or nil activity. We also found that the unfavorable neuroblastoma often have a loss of heterozygosity (LOH) at the MYCL locus. To clarify the biological and clinical profiles of tumors with genetic abnormalities of the short arm of chromosome 1, we performed deletion mapping on 1p on 92 neuroblastoma tissues and corresponding noncancerous samples obtained from 92 cases for 24 micro- or minisatellite loci. LOH was detected in at least one locus of 1p in 43 (47%) cases. All samples were classified into four groups according to the deleted pattern: interstitial deletion (group I, n = 20), short terminal deletion (group ST, n = 6), large terminal deletion (group LT, n = 17), and without detectable deletion (group N, n = 49). All group I cases, whose SRO (shortest region of overlap) was at 1p36.1-2, survived disease free, and none of them showed MYCN amplification or high telomerase activity except for one case. On the other hand, in group LT cases, who showed a large terminal deletion from D1S162 (1p32-pter), including the SRO of group I, only 5 out of 17 have survived disease free, and 13 showed MYCN amplification or high telomerase activity. The six group ST cases showed small terminal deletion from 1p36.3 with modest prognosis, similar to the group N. Thus, we propose three loci, 1p36.1-2, 1p32-34, and 1p36.3, as the candidate loci of neuroblastoma suppressor genes on chromosome 1p responsible for groups I, LT, and ST, respectively. Among them, the 1p32-34 locus may be associated with aggressiveness of tumor progression, possibly due to MYCN amplification and/or telomerase reactivation, while the remaining two loci may not. Med. Pediatr. Oncol. 36: 67–74, 2001. © 2001 Wiley-Liss, Inc.

Telomerase activity in breast fine-needle aspiration (FNA) samples may have diagnostic utility. The purpose of this study was to compare in FNA samples of breast tumor the diagnostic accuracy as correlated with histologic final diagnosis. Fine-needle aspiration samples were obtained from 617 patients with palpable breast tumors. Slide preparation and cytology were performed according to a uniform approach. Extracts derived from 10³ cells from the residual cells in the syringe were used for the telomeric repeat amplification protocol (TRAP) assay. Of the original 617 patients, 220 underwent open biopsy or surgery, and 93 cancers and 127 patients' benign diseases were diagnosed by histologic examination. All 62 tumors that were diagnosed as “malignant” or “probably malignant” by FNA cytology were cancerous, and 50 cases (81%) showed detectable telomerase activity. Among 17 “atypical” or “indeterminate” cases, all 10 tumors with detectable telomerase activity subsequently were diagnosed as breast carcinoma whereas 6 of 7 tumors without telomerase activity were diagnosed as benign. Among the 141 “benign” or “unsatisfactory” samples, 12 of 21 cases with detectable telomerase activity subsequently were diagnosed as cancer. The diagnostic accuracy of telomerase activity in FNA samples is considered to be equivalent or slightly higher to that of cytology (86% vs. 70%). Detection of telomerase activity should be considered an alert for false-negative results of FNA cytology and may be useful as a diagnostic marker for breast malignancy, especially in samples cytologically undetermined to be malignant. Cancer (Cancer Cytopathol) 2000;90:235–8. © 2000 American Cancer Society.

Although multifocal neuroblastoma is rare, its incidence has increased because of recent improvements in diagnostic tools and the introduction of mass screening. Among the 106 neuroblastoma cases treated at the authors' hospital between 1984 and 1998, 8 were multifocal neuroblastoma. The authors examined clinicopathologic findings and biologic features, including MYCN amplification, NTRK1 and Ha-ras p21 expression, cellular DNA content, and telomerase activity in these 8 multifocal neuroblastoma cases. Moreover, clinicopathologic findings were investigated with a review of 53 published cases of multiple neuroblastoma in the literature published in English between 1966 and 1999. Among these eight cases, five were detected by mass screening and three were incidental neuroblastomas. Histologically, all tumors were classified as ganglioneuroma or favorable neuroblastoma except one advanced case. All tumors lacked the MYCN gene amplification and expressed NTRAK1 mRNA and Ha-ras p21 protein. Cellular DNA content showed that half of these tumors were near-triploid, and the proliferative index (%S-phase) of all tumors was less than 25%. High telomerase activity was detected in none of these cases. Four patients underwent multistage operation and five patients with bilateral adrenal neuroblastomas underwent tumor enucleation to preserve adrenal function. Currently, all patients are disease free and none have required corticosteroid replacement therapy. Among the previously reported 53 cases with multifocal neuroblastoma, 25 were incidentally detected, 18 had familiar history, and most patients without other major complications also had extremely good prognoses. These findings suggested that most multifocal neuroblastomas have favorable biologic features. Clinically, surgical approaches should be attempted to preserve organ function, especially adrenal function, and minimal invasive surgery should be performed. In cases of thoracoabdominal neuroblastoma, multistage surgery is effective and safe. Cancer 2000;88:1955–63. © 2000 American Cancer Society.

Direct trapping of a single floating cell, i.e.a white blood cell from a drop of blood, within a nanospray tip was followed by super-sonication after the addition of ionization solvent. Molecular detection of an increased number of peaks with a higher intensity and a wider m/zrange, which extends from metabolites to lipids, was acquired than of that without sonication. This method was applied to a few separated circulating tumor cells (CTC) from a neuroblastoma patient’s blood to obtain their lipido-metabolomic molecular profile at the single cell level. In addition to vital molecules such as amino acids, catechol amine metabolites, which are specific to neuroblastoma, and drugs included in the patient’s course of therapy were detected. This established “direct single-cell lipido-metabolomic method” seems to be useful for direct and wide range molecular detection not only for many live single-cells, but also for rare cells, such as CTCs, for future molecular diagnosis.

KAI1, a metastasis suppressor gene of prostate cancer, is located on human chromosome 11p11.2. Down‐regulation of KAI1mRNA during tumor progression and metastasis has been reported for several kinds of cancer, but the mechanism of this down‐regulation is not known. In the present study, our aim was to ascertain the relationship between down‐regulation of KAI1mRNA expression and KAI1gene alterations in lung cancer. Forty‐nine cases of adenocarcinoma of the lung were studied by reverse‐transcriptase polymerase chain reaction (RT‐PCR) assay of KAI1mRNA and by immunohistochemical detection of KAI1 protein. In addition, markers of the microsatellite loci D11S1344 and D11S1326 were used to investigate loss of heterozygosity (LOH) and replication errors (RERs) of the KAI1gene region. The RT‐PCR assay showed that there was no correlation between KAI1mRNA expression and either the age of the patients or tumor size. By contrast, KAI1mRNA expression was significantly correlated with gender (P=0.047), metastasis to the lymph nodes or other organs (P=0.004), the histological grade of the tumor (P=0.036) and the pathological stage (P=0.049). Immunohistochemical staining showed that in one case without metastasis, loss of KAI1mRNA was associated with invasion of the stroma by KAI1 protein‐negative cancer cells. The numbers of informative cases by microsatellite analysis were 14 (28.6%) of 49 at D11S1344 and 27 (55.1%) of 49 at D11S1326; none of 49 adenocarcinomas showed LOH or RERs at these loci. These results suggest that down‐regulation of KAI1mRNA expression rarely if ever involves LOH or RERs of the KAI1gene region in primary lung adenocarcinoma.

Abstract: We examined the pancreatic tissue concentrations of cefazolin in ten patients undergoing pancreatectomy, and determined the optimal intraoperative time to deliver a repeat dose of cefazolin. An intravenous bolus dose of 1 g cefazolin was administered at the time of skin incision. Peripheral blood, subcutaneous adipose tissue, and peritoneal samples were obtained intraoperatively every hour for 4h after the antibiotic was first administered, and pancreatic tissue samples were obtained at the time of pancreatectomy. To determine adquate tissue levels of cefazolin, minimum inhibitory concentrations (MIC) were measured for four bacterial species, namely 360 isolates of methicillin-sensitiveStaphylococcus aureus (MSSA), 204 isolates ofKlebsiella pneumoniae, 314 isolates ofEscherichia coli, and 30 isolates ofStreptococcus spp. The antibiotic concentrations in adipose tissue and peritoneum 3 h after the administration of cefazolin were lower than the MIC80 forK. pneumoniae, E. coli, andStreptococcus spp. Most pancreatic tissue samples showed antibiotic concentrations greater than the MIC80 for these bacterial species; however, those from four patients complicated by severe chronic pancreatitis, massive intraoperative bleeding, or obesity showed concentrations lower than the MIC80. Thus, we recommend that a second dose of cefazolin be given 3 h after thefirst administration to maintain adequate levels of antibiotic activity.

Abstract: Pancreatic ascites can occur in association with the rupture of a pseudocyst or the disruption of a pancreatic duct during the natural course of chronic pancreatitis. We report herein the successful treatment of three patients with pancreatic ascites by performing a surgical procedure after 4–6 weeks of total parenteral nutrition (TPN) proved ineffective. The principles of our surgical procedure for pancreatic ascites are as follows: (1) minimum pancreatic tissue is resected; (2) surgical intervention to repair leaking sites is not necessary; (3) pancreatic duct drainage is facilitated by an intestinal Rouxen-Y loop; (4) An external drainage tube is inserted through the Roux-en-Y loop into the main pancreatic duct. All three patients who underwent our surgical procedure had a good outcome. Although the mean follow-up time is still only 18.3 months, their condition has improved, with no evidence of recurrent ascites. Thus, our surgical procedure should be considered as an appropriate treatment for pancreatic ascites because it can be applied for all types of leakage, including leakage from the posterior wall of pancreas; it preserves pancreatif function, especially endocrine function; and it enables preservation of the spleen.

Transmembrane proteins of the CD44 family play roles in cell-cell and cell-matrix interactions, and their aberrant expression has been reported to be associated with the growth and metastasis of various tumors. The authors examined CD44 standard (CD44st) and CD44 variant 6 (CD44v6) expression in extrahepatic bile duct (EHBD)/ampullary carcinoma. In 36 EHBD/ampullary carcinomas, immunohistochemical analyses with monoclonal antibodies against the human CD44st protein or CD44v6 protein were performed by the streptavidin-biotin immunoperoxidase method. The relation between expression of the proteins and lymph node metastases or patient outcome was investigated. To verify the lack of CD44v6 mRNA expression in tumors negative for CD44v6 staining, reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot hybridization were performed. Immunohistochemical analyses revealed that 13 of 36 carcinomas (36.1%) expressed the CD44v6 protein. Only 2 of 13 CD44v6-expressing primary tumors (15.4%) had regional lymph node metastases, whereas 14 of 23 tumors showing no CD44v6 expression (60.9%) had lymph node metastases (Fisher exact test, P < 0.01). Moreover, a lack of CD44v6 expression was correlated significantly with poor prognosis (generalized Wilcoxon test, P < 0.05). Eleven of 13 CD44v6 positive tumors showed CD44st expression, which also was correlated with prognosis. RT-PCR and Southern blot hybridization revealed a lack of CD44v6 mRNA expression in the tumors that were negative for CD44v6 staining. The results of the current study suggest that a lack of expression of CD44, especially CD44v6, is correlated with lymph node metastases and poor prognosis, and may be a prognostic factor for patients with EHBD/ampullary carcinoma. Cancer 1999;86:1691–9. © 1999 American Cancer Society.

We examined the clinical features of 14 men (mean age 72 years) with postoperative enteritis caused by methicillin-resistantStaphylococcus aureus(MRSA). The patients had all undergone surgery for the treatment of digestive diseases and had received antibiotic prophylaxis consisting of an extended-spectrum cephem. Diarrhea appeared a mean of 3.3 days postoperatively and lasted for 5 days on average. In severe cases organ insufficiency was involved. Coagulase-positive staphylococci were the predominant organisms isolated from watery diarrhea. In 13 of 14 patients, coagulase type II isolates producing enterotoxins A, C and toxic shock syndrome toxin-1 (TSST-1) withenterotoxin A, C,andtstgenes were isolated. These strains were sensitive to vancomycin and arbekacin; however, they were highly resistant to many other antibiotics. We also investigated the effects of a glucocorticoid hormone and gamma globulin on production of tumor necrosis factor-α (TNF-α) and interleukin-2 (IL-2) obtained from healthy volunteers. TNF-α and IL-2 production was enhanced by TSST-1 and the supernatant of Iscove-modified dulbecco medium, in which coagulase type II isolates producing enterotoxins A, C and TSST-1 with enterotoxin A, C were cultured for 24h. Both glucocorticoid hormone and gama globulin suppressed TNF-α and IL-2 production, thus suggesting that these drugs may be effective in treating postoperative MRSA enteritis.

The association of human telomerase activity with an indefinite replicative capacity of cells in vitro and advanced tumors in vivo is gaining wide support. The increasing interest in studying various aspects of telomerase expression in cancer required the development of a sensitive and reliable protocol for the extraction and detection of telomerase activity in cell culture material, and from small tissue samples obtained from biopsy, surgical reaction of tumors, and autopsy. Recently a novel procedure for the extraction and detection of telomerase activity was developed (Science 1994; 266: 2011–2015) which resulted in an estimated 104 fold improvement in detectability compared with previous methods. The described procedures not only dramatically increased sensitivity but also allowed fast and efficient detection of telomerase activity in a large number of samples. A number of technical aspects which are of eritical importance for reproducibility and reliability of this assay using clinical material are addressed in this report. In addition, new methods to perform telomerase assays without the use of radioisotopes are described.

Telomerase activity was analysed in 100 neuroblastoma cases. Although telomerase activity was not detected in normal adrenal tissues or benign ganglioneuromas, almost all neuroblastomas (94%) did express it, suggesting an important role for telomerase in neuroblastoma development. Neuroblastomas with high telomerase activity had other genetic changes (for example, N-mycamplification) and an unfavourable prognosis, whereas tumours with low telomerase activity were devoid of such genetic alterations and were associated with a favourable prognosis. Three neuroblastomas lacking telomerase activity regressed (stage IVS). Thus telomerase expression may be required as a critical step in the multigenetic process of tumorigenesis, and two different pathways may exist for the development of neuroblastoma.

Telomerase activity is observed in most malignant tumors and germ cells, whereas normal somatic cells usually do not express it. Human endometrium is composed of glandular and stromal components and exhibits dramatic changes in proliferative activity during the menstrual cycle, which is exquisitely regulated by estrogen function. We previously reported that normal human endometrium expresses telomerase activity. However, it remains unclear which of the above components are the major sources of telomerase activity and how levels of telomerase activity are regulated over the menstrual cycle. Quantitative analysis of telomerase activity revealed that it changes dramatically over the course of the menstrual cycle and is strictly regulated in a menstrual-phase-dependent manner. Maximal activity equivalent to that in endometrial cancer was present in late proliferative phase, and minimal activity in late secretory phase. Postmenopausal endometrium and endometrium treated with anti-estrogen drugs exhibited decreased telomerase activity. Testing isolated epithelial glandular cells and stromal cells, we found that telomerase activity was localized to epithelial glandular cells.In situRNA hybridization analysis also revealed epithelial-specific expression of human telomerase RNA. In vitroanalysis of cultured epithelial cells demonstrated that telomerase activity is correlated with epithelial proliferation but not affected by estrogen treatment. These findings suggest that expression of telomerase activity is specific to epithelial cells and linked to cell proliferative status. The involvement of estrogen in telomerase regulation remains to be elucidated.

The severity and predicted outcome of postoperative Pseudomonas aeruginosa (P. aeruginosa) infections (PPAI) was evaluated using a severity scoring system based on a simplification and modification of the APACHE II system. A total of 86 patients in whom P. aeruginosa was isolated from various sources were examined. PPAI developed in 50 patients, resulting in an overall mortality rate of 24%. An increased severity score (SS) correlated with an increased risk of developing PPAI. Thus, PPAI developed in 33% of the patients with an SS of 0–1, in 66.7% of those with an SS of 2–3, and in 100% of those with an SS of 6 or higher. Moreover, the mortality rate of the patients with an initial score of 6 or higher was 50%. The mean (±SD) initial severity score was 5.4±2.9 for survivors and 2.9±2.6 for nonsurvivors (P<0.01). In the patients who subsequently died, the SS remained high throughout the clinical course despite therapy, whereas in the survivors the SS decreased progressively, reflecting a favorable clinical course. These results suggest that our severity scoring system was useful for predicting outcome and monitoring the response of PPAI to therapy.

We herein report a case of paratesticular cholesterol granuloma. A 5-year-old boy with a painless mass (10 mm in diameter) in the left scrotum. Although the mass shrank over the next eight months, it increased in size at six years of age. Surgical resection of the mass was performed. The mass originated from the tunica albuginea. The histological diagnosis was cholesterol granuloma. To our knowledge, this is the first case of cholesterol granuloma in the scrotum of a child.

The CHARGE association, choanal arresia or coloboma with multiple anomalies, is rare. A newborn boy with CHARGE association was referred to our hospital because of bilateral choanal atresia. Additionally, he had left renal aplasia, one of the anomalies associated with this syndrome. A right suprarenal neuroblastoma (stage IV) was diagnosed when the patient was 5 months old. At operation, great care was taken not to injure the right kidney, and resection of the primary tumor as well as radical dissection of the right renal hilar and para-aortic lymph nodes were performed without difficulty. The patient received postoperative chemotherapy with vincristine, cyclophosphamide, and adriamycin for 2 years and is alive without recurrence 5 years after the operation. No report of the CHARGE association with a malignancy has appeared in the literature. The relationship between the CHARGE association and malignant tumors is still obscure, so that further study is necessary to define it.