Search

Among western African countries, the Republic of Ghana has maintained an economic growth rate of 5% since the 1980s and is now categorized as a middle-income country. However, as with other developing countries, Ghana still has challenges in the effective implementation of surveillance for infectious diseases. Facing public health emergencies of international concern (PHEIC), it is crucial to establish a reliable sample transportation system to the referral laboratory. Previously, surveillance capacity in Ghana was limited based on Integrated Disease Surveillance and Response, and therefore the “Surveillance and Laboratory Support for Emerging Pathogens of Public Health Importance in Ghana (SLEP)” was introduced to strengthen diarrhea surveillance. The SLEP project started with a sentinel diarrhea survey supported by SATREPS/JICA in collaboration with National Public Health Reference Laboratory (NHPRL) and Noguchi Memorial Institute of Medicine (NMIMR). The base-line survey revealed the limited capacity to detect diarrhea pathogens and to transfer samples from health centers to NHPRL. The involvement of private clinic/hospital facilities into the surveillance network is also crucial to strengthen surveillance in Ghana. The strong and interactive relationship between the two top referral laboratories, NHPRL under the Ministry of Health NMIMR and under the Ministry of Education, enables Ghana Health Services and is critical for the rapid response against PHEIC. In future, we hope that the outcome of the SLEP surveillance project could contribute to building a surveillance network with more timely investigation and transfer of samples to referral labs.

Background: The 95-95-95 UNAIDS global strategy was adapted to end the AIDS epidemic by 2030. The target is based on the premise that early detection of HIV-infected persons and linking them to treatment regardless of their CD4 counts will lead to sustained viral suppression. HIV testing strategies to increase uptake of testing in Western and Central Africa remain inadequate. Hence, a high proportion of people living with HIV in this region do not know their status. This report describes the implementation of a community-based health screening (also known as “Know Your Status” -KYS), as part of basic science research, in a way that contributed to achieving public health goals. Methods: A community-based health screening was conducted in 7 communities within the Eastern region of Ghana between November 2017 to April 2018, to recruit and match HIV seronegative persons to HIV seropositive persons in a case-control HIV gut microbiota study. Health assessments included blood pressure, body mass index, blood sugar, Hepatitis B virus, syphilis, and HIV testing for those who consented. HIV seronegative participants who consented were consecutively enrolled in an ongoing HIV gut microbiota case-control study. Descriptive statistics (percentages) were used to analyze data. Results: Out of 738 people screened during the exercise, 700 consented to HIV testing and 23 (3%) were HIV positive. Hepatitis B virus infection was detected in 4% (33/738) and Syphilis in 2 % (17/738). Co-infection of HIV and HBV was detected in 4 persons. The HIV prevalence of 3% found in these communities is higher than both the national prevalence of 1.7% and the Eastern Regional prevalence of 2.7 in 2018. Conclusion: Community-based health screening, such as undertaken in our study could be critical for identifying infected persons from the community and linking them to care. This will greatly contribute to achieving the first two 95s and working towards ending AIDS by 2030.

Expanding access to effective antiretroviral therapy (ART) is a major tool for management of Human Immunodeficiency Virus (HIV) infection. However, rising levels of HIV drug-resistance have significantly hampered the anticipated success of ART in persons living with HIV (PLWH), particularly those from Africa. Though great strides have been made in Ghana toward achieving the UNAIDS “95-95-95” target, a substantial number of PLWH receiving ART have not attained viral suppression. This study investigated patterns of drug resistance mutations in ART naïve as well as ART-experienced PLWH receiving first-line regimen drugs from Ghana. In a cross-sectional study, blood samples were collected from HIV-1 infected adults (≥18 years) attending HIV/AIDS clinic at the Eastern Regional Hospital, Koforidua, Ghana from September to October 2017. Viral RNA isolated from plasma were subjected to genotypic drug resistance testing for Protease Inhibitors (PI), Reverse Transcriptase Inhibitors (RTI), and Integrase Strand Transfer Inhibitors (INSTI). A total of 95 (84 ART experienced, 11 ART naïve) HIV-1 infected participants were sampled in this study. Sixty percent (50/84) of the ART-experienced participants were controlling viremia (viral load

An attenuated mutant strain of melon yellow spot virus (MYSV), an Orthotospovirus, designated as SA08-8, was obtained from a pathogenic isolate, C95S, via high temperature treatment, single local lesion isolation, and serial mechanical inoculation. Under greenhouse conditions, the SA08-8 strain conferred the cross-protection against wild-type MYSV but it was not transmissible by its known natural vector melon thrips. In SA08-8 genome, a total of 16 amino acid substitutions were found in the genes encoding RNA-dependent RNA polymerase, glycoprotein precursor, and nonstructural S protein. This attenuated mutant strain is a potential biocontrol agent for MYSV-induced crop diseases in the field.

ASEAN realized the ASEAN Economic Community (AEC) at the end of 2015. The AEC is an economic integration whose goals are free movement of goods, services, investment, and skilled workers, and freer movement of capital. Following the establishment of the ASEAN Free Trade Area (AFTA), ASEAN deepened its economic integration through the AEC. Evaluations of ASEAN’s economic integration are mixed. While highly praised for its steady integration progress, there are also assessments suggesting there is little progress in integration. This study examines the AEC’s economic integration and evaluates ASEAN economic integration, including the AFTA. The AEC is an “FTA-plus” economic integration, and its target and level of market integration are similar to those of the Economic Partnership Agreement. The AEC’s greatest achievement is using tariff elimination to realize a free trade area with a high level of trade liberalization. Liberalizing trade in services, eliminating non-tariff barriers, and facilitating trade have been delayed and have become issues for AEC2025. AEC2025 aims to participate in the global value chain, and its measure is the attraction of foreign direct investment. Although ASEAN’s economic integration has various issues, as a whole, it can be evaluated as successful because of its implementation of flexible liberalization over time. ASEAN’s economic integration is a successful example of economic integration by developing countries; other developing countries can learn lessons from ASEAN’s experiences.

Diarrheal disease remains a major global health problem particularly in children under 5 years and the emergence of antibiotic-resistant strains of causative pathogens could slow control efforts, particularly in settings where treatment options are limited. This surveillance study conducted in Ghana aimed to determine the prevalence and antimicrobial susceptibility profile of diarrhea-causing bacteria. This was a cross-sectional study carried out in five health facilities in the Ga West Municipality of Ghana between 2017 and 2021. Diarrheic stool samples from patients were collected and cultured on standard differential/selective media and isolates identified by standard biochemical tests, MALDI-TOF assay, and serological analysis. The antibiogram was determined using Kirby-Bauer disk diffusion and Microscan autoScan4 MIC panels which were used for extended-spectrum beta-lactamase (ESBL) detection. Bacteria were isolated from 97.5% (772/792) of stool samples, and 167 of the isolates were diarrheagenic and met our inclusion criteria for antimicrobial resistance (AMR) analysis. These included Escherichia coli (49.1%, 82/167), Salmonella species (23.9%, 40/167), Vibrio species (16.8%, 28/167), and Shigella species (10.2%, 17/167). Among 24 Vibrio species, we observed resistances to cefotaxime (21/24, 87.5%), ceftriaxone (20/24, 83.3%), and ciprofloxacin (6/24, 25%), including four multi-drug resistant isolates. All 13 Vibrio parahaemolyticus isolates were resistant to cefazolin. All 17 Shigella isolates were resistant to tetracycline with resistance to shigellosis drugs such as norfloxacin and ciprofloxacin. Salmonella isolates were highly susceptible to norfloxacin (40/40, 100%) and tetracycline (12/34, 35%). Two ESBL-producing E. coli were also identified with marked susceptibility to gentamicin (66/72, 91.7%) and amikacin (57/72, 79.2%) prescribed in the treatment of E. coli infections. This study showed the different bacteria implicated in diarrhea cases in Ghana and the need for differential diagnoses for better treatment outcomes. Escherichia coli, Shigella, Salmonella, and Vibrio have all been implicated in diarrhea cases in Ghana. The highest prevalence was E. coli and Salmonella with Shigella the least prevalent. Resistance to commonly used drugs found in these isolates may render bacteria infection treatment in the near future nearly impossible. Routine antimicrobial susceptibility testing, effective monitoring, and nationwide surveillance of AMR pathogens should be implemented to curb the increase of antimicrobial resistance in Ghana.

Accurate monitoring of epidemics is a key strategy for controlling human immunodeficiency virus type-1 (HIV-1) infection. To delineate the characteristics of newly diagnosed cases of HIV-1 infection, we assessed the proportion of recent HIV-1 infections using a recent infection-testing algorithm (RITA). In 2015, 248 cases were newly diagnosed with HIV infection at the Regional Hospital Koforidua, Ghana. Of these, 234 cases (94.4%) were infected with HIV-1 only, four (1.6%) were infected with HIV-2 only, and 10 (4.0%) were co-infected with HIV-1 and HIV-2. All HIV-1 single-seropositive samples were used in the HIV-1 LAg avidity assay for RITA. Our analysis revealed that 18 cases (7.7%) were recently infected, indicating that early diagnosis was not achieved in Ghana. This is the first report to assess the proportion of recent infections in Ghana using a biomarker approach. The accumulation of these data will contribute to the accurate estimation of HIV-1 incidence and prevalence in Ghana.

Polymorphisms in human leukocyte antigen (HLA) class I loci are known to have a great impact on disease progression in HIV-1 infection. Prevailing HIV-1 subtypes and HLA genotype distribution are different all over the world, and the HIV-1 and host HLA interaction could be specific to individual areas. Data on the HIV-1 and HLA interaction have been accumulated in HIV-1 subtype B- and C-predominant populations but not fully obtained in West Africa where HIV-1 subtype CRF02_AG is predominant. In the present study, to obtain accurate HLA typing data for analysis of HLA association with disease progression in HIV-1 infection in West African populations, HLA class I (HLA-A, -B, and -C) four-digit allele typing was performed in treatment-naïve HIV-1 infected individuals in Ghana (n = 324) by a super high-resolution single-molecule sequence-based typing (SS-SBT) using next-generation sequencing. Comparison of the SS-SBT-based data with those obtained by a conventional sequencing-based typing (SBT) revealed incorrect assignment of several alleles by SBT. Indeed, HLA-A*23:17, HLA-B*07:06, HLA-C*07:18, and HLA-C*18:02 whose allele frequencies were 2.5%, 0.9%, 4.3%, and 3.7%, respectively, were not determined by SBT. Several HLA alleles were associated with clinical markers, viral load and CD4+ T-cell count. Of note, the impact of HLA-B*57:03 and HLA-B*58:01, known as protective alleles against HIV-1 subtype B and C infection, on clinical markers was not observed in our cohort. This study for the first time presents SS-SBT-based four-digit typing data on HLA-A, -B, and -C alleles in Ghana, describing impact of HLA on viral load and CD4 count in HIV-1 infection. Accumulation of these data would facilitate high-resolution HLA genotyping, contributing to our understanding of the HIV-1 and host HLA interaction in Ghana, West Africa.

Acute gastroenteritis associated with diarrhea is considered a serious disease in Africa and South Asia. In this study, we examined the trends in the causative pathogens of diarrhea and the corresponding gut microbiota in Ghana using microbiome analysis performed on diarrheic stools via 16S rRNA sequencing. In total, 80 patients with diarrhea and 34 healthy adults as controls, from 2017 to 2018, were enrolled in the study. Among the patients with diarrhea, 39 were norovirus-positive and 18 were rotavirus-positive. The analysis of species richness (Chao1) was lower in patients with diarrhea than that in controls. Beta-diversity analysis revealed significant differences between the two groups. Several diarrhea-related pathogens (e.g., Escherichia-Shigella, Klebsiella and Campylobacter) were detected in patients with diarrhea. Furthermore, co-infection with these pathogens and enteroviruses (e.g., norovirus and rotavirus) was observed in several cases. Levels of both Erysipelotrichaceae and Staphylococcaceae family markedly differed between norovirus-positive and -negative diarrheic stools, and the 10 predicted metabolic pathways, including the carbohydrate metabolism pathway, showed significant differences between rotavirus-positive patients with diarrhea and controls. This comparative study of diarrheal pathogens in Ghana revealed specific trends in the gut microbiota signature associated with diarrhea and that pathogen-dependent dysbiosis occurred in viral gastroenteritis.

In human immunodeficiency virus type-1 (HIV-1) infections, cytotoxic T-lymphocyte (CTL) responses targeting human leukocyte antigen (HLA)-restricted viral epitopes exert strong suppressive pressure on viral replication and frequently select for mutations resulting in viral escape from CTL recognition. Numerous data on these HLA-associated mutations in HIV-1 subtypes B and C have been amassed with few reports described in other subtypes. In the present study, we investigated the HLA-associated mutations in HIV-1 subtype CRF02_AG prevailing in Ghana, Western Africa. We determined viral gag sequences in 246 out of 324 HIV-1-infected Ghanaians. Phylogeny analysis revealed that 200 (81.3%) individuals were infected with HIV-1 CRF02_AG. Full gag and vif sequences were obtained from 199 and 138, respectively, out of the 200 individuals infected with CRF02_AG and subjected to determination of HLA-associated mutations. The analysis found HLA-associated HIV-1 CRF02_AG non-synonymous polymorphisms at 19 sites; 13 in gag and six in vif, including those that were newly determined. Generation of this data is an important contribution to our understanding of HIV-1 CRF02_AG and host T cell interaction.

HIV-1 infected individuals under antiretroviral therapy can control viremia but often develop non-AIDS diseases such as cardiovascular and metabolic disorders. Gut microbiome dysbiosis has been indicated to be associated with progression of these diseases. Analyses of gut/fecal microbiome in individual regions are important for our understanding of pathogenesis in HIV-1 infections. However, data on gut/fecal microbiome has not yet been accumulated in West Africa. In the present study, we examined fecal microbiome compositions in HIV-1 infected adults in Ghana, where approximately two-thirds of infected adults are females. In a cross-sectional case-control study, age- and gender-matched HIV-1 infected adults (HIV+; n = 55) and seronegative controls (HIV-; n = 55) were enrolled. Alpha diversity of fecal microbiome in HIV+ was significantly reduced compared to HIV- and associated with CD4 counts. HIV+ showed reduction in varieties of bacteria including Faecalibacterium, the most abundant in seronegative controls, but enrichment of Proteobacteria. Ghanaian HIV+ exhibited enrichment of Dorea and Blautia; bacteria groups whose depletion has been reported in HIV-1 infected individuals in several other cohorts. Furthermore, HIV+ in our cohort exhibited a depletion of Prevotella, a genus whose enrichment has recently been shown in men having sex with men (MSM) regardless of HIV-1 status. The present study revealed the characteristics of dysbiotic fecal microbiome in HIV-1 infected adults in Ghana, a representative of West African populations.

Recent studies have indicated an association between gut microbiome composition and various disorders, including infectious diseases. The composition of the microbiome differs among ethnicities and countries, possibly resulting in diversified interactions between host immunity and the gut microbiome. Characterization of baseline microbiome composition in healthy people is an essential step for better understanding of the biological interactions associated with individual populations. However, data on the gut/fecal microbiome have not been accumulated for individuals in West Africa. In the present study, we examined the fecal microbiome composition in healthy adults in Ghana. Toward this, 16S rRNA gene libraries were prepared using bacterial fractions derived from 55 Ghanaian adults, which were then subjected to next-generation sequencing. The fecal microbiome of the Ghanaian adults was dominated by Firmicutes (Faecalibacterium, Subdoligranulum, and Ruminococcaceae UCG-014), Proteobacteria (Escherichia-Shigella and Klebsiella), and Bacteroidetes (Prevotella 9 and Bacteroides), consistent with previous observations in African cohorts. Further, our analysis revealed differences in microbiome composition and a lower diversity of the fecal microbiome in the Ghanaian cohort compared with those reported in non-African countries. This is the first study to describe substantial fecal microbiome data obtained using high-throughput metagenomic tools on samples derived from a cohort in Ghana. The data may provide a valuable basis for determining the association between the fecal microbiome and progression of various diseases in West African populations.

Purpose. There have been hardly any reports on the human immunodeficiency virus type 1 (HIV-1) drug-resistance profile from northern Ghana since antiretroviral therapy (ART) was introduced over a decade ago. This study investigated prevailing HIV-1 subtypes and examined the occurrence of drug resistance in ART-experienced patients in Tamale, the capital of the Northern Region of Ghana. Methodology. A cross-sectional study was carried out on HIV-infected adult patients receiving first-line ART. HIV viral load (VL) and CD4+ T-cell counts were measured. The pol gene sequences were analysed for genotypic resistance by an in-house HIV-1 drug-resistance test; the prevailing HIV-1 subtypes were analysed in detail. Results/Key findings. A total of 33 subjects were studied. Participants comprised 11 males (33.3 %) and 22 (66.7 %) females, with a median age of 34.5 years [interquartile range (IQR) 30.0–40.3]. The median duration on ART was 12 months (IQR 8.0–24). Of the 24 subjects successfully genotyped, 10 (41.7 %) viruses possessed at least one mutation conferring resistance to nucleoside or non-nucleoside reverse-transcriptase inhibitors (NRTIs/NNRTIs). Two-class drug resistance to NRTI and NNRTI was mostly detected (25 %, 6/24). The most frequent mutations were lamivudine-resistance M184V and efavirenz/nevirapine-resistance K103N. HIV-1 subtype CRF02_AG was predominant (79.2 %). Other HIV-1 subtypes detected were G (8.3 %), A3 (4.2 %) and importantly two (8.3 %) unique HIV-1 recombinant forms with CRF02_AG/A3 mosaic. Conclusion. HIV-1 shows high genetic diversity and on-going viral genetic recombination in the study region. Nearly 42 % of the patients studied harboured a drug-resistant virus. The study underscores the need for continued surveillance of HIV-1 subtype diversity; and of drug-resistance patterns to guide selection of second-line regimens in northern Ghana.

In human immunodeficiency virus type-1 (HIV-1) infections, cytotoxic T-lymphocyte (CTL) responses targeting human leukocyte antigen (HLA)-restricted viral epitopes exert strong suppressive pressure on viral replication and frequently select for mutations resulting in viral escape from CTL recognition. Numerous data on these HLA-associated mutations in HIV-1 subtypes B and C have been amassed with few reports described in other subtypes. In the present study, we investigated the HLA-associated mutations in HIV-1 subtype CRF02_AG prevailing in Ghana, Western Africa. We determined viral gag sequences in 246 out of 324 HIV-1-infected Ghanaians. Phylogeny analysis revealed that 200 (81.3%) individuals were infected with HIV-1 CRF02_AG. Full gag and vif sequences were obtained from 199 and 138, respectively, out of the 200 individuals infected with CRF02_AG and subjected to determination of HLA-associated mutations. The analysis found HLA-associated HIV-1 CRF02_AG non-synonymous polymorphisms at 19 sites; 13 in gag and six in vif, including those that were newly determined. Generation of this data is an important contribution to our understanding of HIV-1 CRF02_AG and host T cell interaction.

BackgroundAn outbreak of novel coronavirus (SARS-CoV-2)-associated respiratory infectious diseases (COVID-19) emerged in 2019 and has spread rapidly in humans around the world. The demonstration of in vitro infectiousness of respiratory specimens is an informative surrogate for SARS-CoV-2 transmission from patients with COVID-19; accordingly, viral isolation assays in cell culture are an important aspect of laboratory diagnostics for COVID-19.MethodsWe developed a simple and rapid protocol for isolating SARS-CoV-2 from respiratory specimens using VeroE6/TMPRSS2 cells, a cell line that is highly susceptible to the virus. We also investigated a correlation between isolation of SARS-CoV-2 and viral load detected by real-time RT-PCR (rRT-PCR) using N2 primer/probe set that has been developed for testing of COVID-19 in Japan.ResultsThe SARS-CoV-2 isolation protocol did not require blind passage of inoculated cells and yielded the results of viral isolation within 7 days after inoculation. Specimens with cycle threshold (Ct) values of 35 were virus isolation-positive, indicating that low viral loads (high Ct values) in upper respiratory specimens do not always indicate no risk of containing transmissible virus.ConclusionIn combination with rRT-PCR, the SARS-CoV-2 isolation protocol provides a means for assessing the potential risk of transmissible virus in upper respiratory specimens.

Resistance to the quinone outside inhibitor (QoI) fungicides in the tea gray blight-causing fungus Pestalotiopsis longiseta is a serious problem in Japanese tea cultivation. We conducted a population genetic analysis of QoI-resistant P. longiseta isolates on the Makinohara Plateau, Shizuoka Prefecture, Japan’s largest tea-growing area, to elucidate the disease’s epidemiology and the spread of QoI resistance. Inter simple sequence repeat (ISSR) analysis of 1,083 isolates from 395 fields collected from 2009 to 2012 detected 42 ISSR types, designated as PL01 to PL42. A total of 18, seven, and 38 ISSR types were detected in highly resistant, moderately resistant, and sensitive isolates, respectively. No distinct phylogenetic relationship corresponding to QoI sensitivity or sampling location was observed. No annual changes in the population genetic structure of highly resistant isolates were observed during the study period. A different ISSR type was predominant among QoI-resistant isolates in each region. Analysis of molecular variance revealed significant genetic differentiation in populations of highly resistant isolates among regions (FCT = 0.213) and farmers (FCT = 0.071). Consequently, we speculate that QoI-resistant P. longiseta strains occurred in a number of clonal lineages and spread by both natural and artificial transmission, such as rain splash and plucking machines, throughout each region on the Makinohara Plateau.

Numbers of HLA-associated polymorphisms have been reported on HIV-1 subtypes B and C, but few on other subtypes. Here, we analyzed HLA-associated gag and nef polymorphisms in HIV-1 subtype A/E prevalent in Vietnam. We determined HLA-A, B and C genotypes in 179 HIV-1-infected Vietnamese by next generation sequencing and analyzed proviral genome sequences in 144 of them, showing that 142 of the 144 were subtype A/E. Analysis revealed HLA-associated subtype A/E gag and nef polymorphisms at nineteen residues including those newly determined. Accumulation of these data would contribute to our understanding of HIV-1 subtype A/E and host immune interaction.

Antiretroviral therapy (ART) and drug resistance studies worldwide have focused almost exclusively on human immunodeficiency virus type 1 (HIV-1). As a result, there is limited information on ART and drug resistance in HIV-2 patients. In Ghana, the HIV epidemic is characterized by the domination of HIV-1, with cocirculating HIV-2. We, therefore, sought to determine viral load and drug resistance mutations in HIV-2 patients to inform the clinical management of such individuals in Ghana. We used purposive sampling to collect blood from 16 consented patients, confirmed as HIV-2 or HIV-1/2 dual infections by serology. A 2-step real-time RT-PCR assay was used to determine plasma HIV-2 RNA viral loads. For drug resistance testing, nucleic acids were extracted from plasma and peripheral blood mononuclear cells. The reverse transcriptase and protease genes of HIV-2 were amplified, sequenced and analyzed for drug resistance mutations and HIV-2 group. HIV-2 viral load was detected in 9 of 16 patients. Six of these had quantifiable viral loads (range: 2.62–5.45 log IU/mL) while 3 had viral loads below the limit of quantification. Sequences were generated from 7 out of 16 samples. Five of these were classified as HIV-2 group B and 2 as HIV-2 group A. HIV-2 drug resistance mutations (M184V, K65R, Y115F) were identified in 1 patient. This study is the first to report HIV-2 viral load and drug resistance mutations in HIV-2 strains from Ghana. The results indicate the need for continuous monitoring of drug resistance among HIV-2- infected patients to improve their clinical management.

Research on Olpidium virulentus, the vector of the serious lettuce big-vein disease, is difficult because its resting spores persist in soil and retain virus for years and the fungus is not culturable. We developed protocols to obtain purified, viable resting spores from infected roots using an enzymatic treatment and two-step density gradient centrifugation and to generate a polyclonal antibody against these spores. The antibody was species-specific in a direct immunostaining assay and in western blots produced two specific bands (ca. 30.5, 29.0 kDa) against resting spores. The detection limit for resting spores in an indirect ELISA was 200 spores/100 μL.

We improved a multiple rapid immunofilter paper assay (multi-RIPA) to simultaneously detect three major viruses of cucumber, Cucumber mosaic virus, Melon yellow spot virus and Zucchini yellow mosaic virus. When we increased the concentration of bovine serum albumin (BSA) in the extraction buffer, a positive band for each virus developed in the multi-RIPA, regardless of the order that the three sensitized white latex particles were immobilized on the strip. The increase in BSA concentration also strengthened the sensitivity of each RIPA. In addition, using a mixture of commercially available, red and blue, sensitized latex solutions as a tracer, purple bands could be produced on the strips. Because each virus yielded an unique color for a positive reaction, the three viruses were easy to distinguish.

Little has been reported on routine prophylactic abdominal drainage after gastrectomy, especially after laparoscopy-assisted distal gastrectomy (LADG). We conducted this retrospective study on patients undergoing LADG to evaluate the benefit of routine drainage in LADG procedures.The subjects were 21 patients who underwent surgery for early gastric cancer (EGC) between January 2004 and March 2008. They comprised 10 who underwent LADG with drainage before January 2006 and 11 who underwent LADG without drainage after February 2006. We compared patient and tumor characteristics, operative results, and postoperative outcomes between the groups.The no-drain group of patients were able to eat their first meal significantly sooner than the drain group patients (P0.01); however, the time to start ambulating, passing flatus, and drinking was similar in the two groups. There were no significant differences between the groups in the postoperative complication rate or the postoperative hospital stay. The drain did not seem to add benefit, and no complications due to the lack of drain placement were noted in the no-drain group.Routine prophylactic abdominal drainage after LADG for EGC may not be necessary.

Aim: As techniques in laparoscopic cholecystectomy (LC) have improved, the role of routine prophylactic abdominal drainage may be limited. A retrospective review was carried out of patients undergoing elective LC to evaluate the benefit of routine drainage in simple uncomplicated procedures. Methods: This study of 295 patients with cholecystolithiasis or gallbladder polyp included 145 patients who underwent LC with drainage and 150 patients who underwent LC without drainage between 2003 and 2007. Allocation to drain or not to drain was non-randomized and based on surgeon preference according to intraoperative findings. Patient characteristics, operative results, and postoperative outcomes were compared between the two groups with univariate analysis. Results: Time to first flatus and length of postoperative hospital stay in the LC without drainage group were shorter than in the LC with drainage group. There was no significant difference between the two groups with respect to postoperative complication rate. No complications were noted due to the lack of drain placement. Conclusion: The use of drain after simple elective uncomplicated LC could safely be limited to appropriate patients as judged by the operating surgeon.

Disruption of the blood-brain barrier (BBB) and/or the blood-cerebrospinal fluid barrier (BCSFB) is thought to be one of the major pathophysiological consequences of meningitis and contributes to the development of adverse neurological outcomes. In order to clarify this hypothesis further, we sequentially quantified the permeability of these barriers with magnetic resonance imaging (MRI) contrast enhancement using gadolinium-diethylene triamine pentaacetic acid (Gd-DTPA) in rats with various experimentally-induced meningitis. Meningeal inflammation was elicited by an intracisternal injection of interleukin (IL)-1beta, prostaglandin (PG) E(2), or lipopolysaccharide (LPS). Barrier permeability was calculated from the gadolinium-enhancement ratio (GER) in the subarachnoid space (SAS). The secretion of Gd-DTPA into the SAS was monitored by T1-weighted imaging after an intravenous injection of Gd-DTPA. As a significant linear correlation was observed between the GER and the standard solution, the concentration of the secreted Gd-DTPA were determined from the GER. The maximal intensity in SAS was detected at 5min after Gd-DTPA administration and it declined gradually. Among the inflammatory agents, IL-1beta was found to induce the most severe meningitis as determined from the GER. The concentration of Gd-DTPA in the SAS increased in a dose-dependent manner following IL-1beta intracisternal injection. On the other hand, no significant changes in signal intensity of the brain parenchymal areas due to IL-1beta injection were observed. The findings suggest that the permeability of the BCSFB can be evaluated quantitatively by calculating the GER. MRI with Gd-DTPA provides a useful method to monitor the change in the permeability to the brain barriers.

793 Background: We have reported the effectiveness of neoadjuvant radiotherapy (RT) combined with S-1 in terms of compliance and pathologic response for locally advanced rectal cancer. In the present study, we aimed to evaluate the safety and efficacy of laparoscopic surgery for locally advanced rectal cancer following neoadjuvant RT combined with S-1 in comparison with open surgery. Methods: Two multicenter prospective phase II trials were conducted (UMIN003396, UMIN003398). A total of 63 patients with locally advanced rectal cancer classified as T3-4, N0-3, and M0 were enrolled, and neoadjuvant RT combined with S-1 and total mesorectal excision with D3 lymphadenectomy was performed. Of these, 57 patients were analyzed and divided into a laparoscopic group (LAP, n = 43) and open group (OP, n = 14). We evaluated the short- and long-term outcomes of laparoscopic surgery compared with open surgery by univariate and multivariate analyses. Results: In the patient background, there were no significant differences between the two groups except that cases with T3 and N0 were significantly higher in the LAP compared with the OP. In the operative findings, operation time was longer (mean 447 min vs. 352 min, p = 0.007) and blood loss was lesser (median 220 ml vs. 485 ml, p = 0.033) in the LAP than those in the OP. Although there were no significant differences observed in the incidence of perioperative and late complications between the two groups, reoperation within 30 days was significantly less in the LAP compared with the OP (1 case vs. 5 cases, p = 0.0004). In the multivariate analysis, a distance of the tumor from anal verge within 3 cm was the independent risk factor for reoperation within 30 days. Furthermore, estimated 5-year disease-free survival (LAP 72 % vs. OP 73 %, p = 0.945) and 5-year overall survival (LAP 76 % vs. 75 %, p = 0.836) didn’t significantly differ between the two groups in the Kaplan–Meier curve. Conclusions: The findings of this study demonstrated that laparoscopic surgery for locally advanced rectal cancer following neoadjuvant RT combined with S-1 could be an optional procedure in terms of short- and long-term outcomes. Clinical trial information: UMIN000003396, UMIN000003398.

Sensitivity of the novolac positive resist THMR iP-3300 to violet and blue light emitting diodes with central wavelengths of 404-470 nm was investigated to utilize for developing a new numbering exposure system using a matrix array of such light emitting diodes. In addition, the spectral transmittance of the resist was measured, and the relationship between transmittance and sensitivity was discussed. It was clarified that the sensitivity greatly differed between exposures to violet and blue light rays as much as more than 30 times, and the resist was sensitized only by 3 s exposure to the violet light at minimum. Therefore, reasonable exposure time performance will be obtained if the relevant reduction projection lens is applied.

A case of retroperitoneal mucinous cystadenoma treated laparoscopically is reported. A 26-year-old woman, who complained of abdominal fullness, was referred to us after a large intra-abdominal mass was discovered at her local hospital. Ultrasound and computed tomography showed a fluid-filled mass, 9 cm in diameter, in the left retroperitoneal space. Laparoscopic surgery was performed. We found a smooth and thin-walled cystic tumor that displaced the descending colon to the right and arose from the retroperitoneum, loosely adhering to the psoas muscle. We successfully extirpated the tumor laparoscopically. The surgical specimen consisted of a multilocular, red to yellowish-gray, cystic mass with a smooth, glistening external surface, and inner lining. The mass was filled with thin, yellowish fluid. The histologic diagnosis was benign mucinous cystadenoma. The postoperative course was uneventful, and the patient remains free of recurrence 15 months after surgery. Retroperitoneal mucinous cystadenoma is a rare entity, and the treatment of choice is surgical excision. Laparoscopic extirpation should be considered for the treatment of selected retroperitoneal cystic lesions.

Objective Many studies have described the impact of population ageing on health care expenditures, but few have assessed its impact on specific diseases adjusted for severity and procedure. This study examined the relationship between an ageing population and resource use in patients with cerebrovascular disease (CVD). Methods A total of 13,856 CVD patients were treated in 82 academic and 92 community hospitals. Demographic variables, clinical variables, length of stay (LOS) and total charges were analysed by age group (under 65 years, 65-74 years and 75 years or older). The independent effects of age on LOS and total charge were determined using multivariate analysis. Results There were 5172 (37%) patients under 65 years of age, 4096 (30%) 65-74 years and 4588 (33%) 75 years or older. Intracranial infarction or ischaemia was diagnosed in 69% of the patients, haemorrhage in 23% and subarachnoid haemorrhage in 9%. The overall mortality was 6% (5% in under 65 years, 5% in 65-74 years and 9% in 75 years or older; P < 0.001). There were significant differences in the proportion of procedures performed in each age category. Age and procedure were significantly associated with LOS, particularly the latter. Age had no significant association with total charge, but procedure was highly associated. Conclusions Ageing has no significant impact on total charge. Instead policy-makers should acknowledge the effect of procedures on health care costs, conduct economic evaluations and monitor use of procedures.

High temperature deformation behavior of AZ31 and AZ91 magnesium alloys was examined by compression tests over a wide strain rate range from 10–3 to 103 s–1 with emphasis on the behavior at high strain rates. The dominant deformation mechanism in the low strain rate range below 10–1 s–1 was suggested to be climb-controlled dislocation creep. On the other hand, experimental results indicated that the deformation at a high strain rate of ~103 s–1 proceeds by conventional plastic flow of dislocation glide and twinning even at elevated temperatures. The solid-solution strengthening was operative for high temperature deformation at ~103 s–1.

Differential speed rolling (DSR) was applied to the AZ31 magnesium alloy, intended to modify the texture and thus to enhance the room temperature ductility. Especially, effect of DSR temperature on resulting room temperature tensile properties and texture was investigated at a fixed rolling speed ratio of 1.1. The strength was not affected by the rolling temperature: the yield strength and the ultimate tensile strength for all materials processed by DSR were approximately 240 and 290 MPa, respectively. On the other hand, elongation-to-failure increased from 13.6 to 18.5% with decreasing rolling temperature from 573 to 473 K. In addition, the material processed by DSR exhibited approximately 1.5 times larger ductility than that by conventional symmetric rolling at a rolling temperature of 473 K. The DSR at 523 and 473 K resulted in the basal plane orientation at the position inclined by ∼5 and 8° from the normal direction toward the rolling direction, respectively. It was suggested that the higher ductility of the material processed at lower temperatures is attributed to the slight change in the basal plane orientation and/or reduction in grain size.

Serologic screening for antibodies to the acquired immunodeficiency syndrome (AIDS) virus human immunodeficiency virus (HIV) has been carried out in Ghana since 1986 when the 1st AIDS case was detected. In general surveys have indicated a low HIV-1 seroprevalence rate among the general population. Of 1852 blood donors screened in 1986 only 2 (0.1%) were positive. None of 168 laboratory workers screened at the same time was seropositive. Also demonstrated has been a relatively low (2.1%) seroprevalence rate among local prostitutes in Accra. However of 151 Ghanaian women who worked as prostitutes in the Ivory Coast 49% were HIV-1 seropositive upon their return to Ghana. Similarly 215 males and females from the general population who had spent time living in the Ivory Coast and other West African countries such as Burkina Faso were found to be seropositive. In all 212 Ghanians have been tested as seropositive for HIV infection in the March 1986-July 1987 period. The greatest number of cases have been reported from Ghanas Eastern Region Greater Accra and Ashanti. The male to female ratio of HIV-positive cases was 1 to 11.6 in 1986 and 1 to 6.3 in 1987. The predominant presenting complaints of AIDS patients in Ghana are weight loss (89%) persistent or intermittent diarrhea (78%) respiratory symptoms (67%) fever and night sweats (56%) and central nervous system symptoms (50%). It is believed that more than 50% of prostitutes in the Ivory Coast are from Ghana. Many of these women return to their home country for holidays or to seek medical care when they develop symptoms of HIV infection. Unless effective intervention is implemented to limit the heterosexual spread of AIDS in Ghana its incidence may soon become as high as it is in Central Africa.

In gene expression analyses using a high-density oligonucleotide array in a rat ischemia model, two comparison methods, "pair-wise comparison" and "sample average comparison", were evaluated based on statistical methods. The reliability of the elements screened with a 1.2 to 10-fold threshold was also evaluated. In pair-wise comparisons, most of the elements were significantly independent of the threshold value, with the percentage of significant elements remaining above 95%, when screened at 2.5-fold or higher threshold value. Pair-wise comparison structurally provided strict screening, which resulted in genes that were not selected despite significant alterations in expression. Screening by "sample average comparison" resulted in elements with low probability of significance, which suggested the necessity for increasing the reliability by additional statistical analyses after screening. When genes with altered expression were screened using an oligonucleotide array, marked differences in the numbers and reliability were proved to exist among elements screened by each sample comparison method.

Lead (Pb) is one of the most effective alloying elements to improve the machinability of steels. However the movement of eliminating Pb contents is recently increasing because of reducing environmental impact. Therefore new types of free cutting steels without Pb are strongly required. Ti4C2S2 is one of the promissing candidates of free cutting elements. However the mechanism of improvement in machinability by Ti4C2S2 has not been confirmed yet. In this study we investgated the effect of Ti4C2S2 in 13 % Cr stainless steels. Ti4C2S2 had notch effect like other sulfides such as MnS. Thereby chip breakability was improved. Ti4C2S2 also had tool-protecting effect by forming stable adhesion. On the other hand Ti4C2S2 was not shown remarkable lubricant effect as Pb. The improvement of machinability in 13 mass%Cr ferritic stainless steels by including Ti4C2S2 attribute to the chip breakability and restraint of the tool-wear by forming stable adhesion.

The microstructures and mechanical properties of fullerene powder dispersed AZ91 magnesium alloy composites were examined. The AZ91 with 0, 1 and 5 mass% fullerene additions were produced by extruding a mixture of AZ91 machined chips and fullerene powder. The matrix grains of the extruded materials were equiaxed, and the size was � 5 mm irrespective of the fullerene content. The agglomerated fullerene powder was aligned parallel to the extrusion direction in the composites. The addition of 1 mass% fullerene had little effect on the elastic moduli, tensile strength and ductility, whereas the addition of 5 mass% fullerene slightly decreased them. A comparison between the tensile properties of the present composites and those of AZ91 alloys processed by various methods was also made. The damping capacity increased steeply by the fullerene addition. The effectiveness of the fullerene addition for the enhancement of the damping capacity of metallic alloys was verified. A good combination of damping capacity, elastic moduli, strength and ductility was attained in the present composites.

The human immunodeficiency virus type-1 (HIV-1)-encoded vif protein is essential for viral replication, virion production, and pathogenicity. HIV-1 Vif interacts with the endogenous human APOBEC3G protein (an mRNA editor) in target cells to prevent its encapsidation into virions. Some studies have established targets within the HIV-1 vif gene that are important for its biologic function; however, it is important to determine effective therapeutic targets in vif because of its critical role in HIV-1 infectivity and pathogenicity. The present study demonstrates that virions generated in transfected HeLa-CD4+ cells, especially from HIV-1 vif frame-shift mutant (3' delta vif; 5561-5849), were affected in splicing and had low infectivity in MT-4 cells. In addition, HIV-1 vif antisense RNA fragments constructed within the same region, notably the region spanning nucleic acid positions 5561-5705 (M-3'-AS), which corresponds to amino acid residues 96-144, significantly inhibited HIV-1 replication in MT-4 and reduced the HIV-1 vif mRNA transcripts and reporter gene (EGFP) expression. The generated virions showed low secondary infection in H9 cells. These data therefore suggest that the middle to the 3' end of vif is important for its biological activity in the target cells.

Liver metastasis of colorectal malignancies is an important prognostic factor. Several studies have demonstrated that carbon dioxide (CO2) pneumoperitoneum enhances liver metastasis in animal models. Little is known about intercellular adhesion molecule-1 (ICAM-1) and tumor necrosis factor-alpha (TNF-(alpha) mRNA expression in the liver after CO2 pneumoperitoneum.Forty-five male BALB/c mice were randomly divided into three groups after intra-splenic tumor cell (colon 26) inoculation and the following procedures were performed: CO2 pneumoperitoneum (n = 15), open laparotomy (n = 15), and anesthesia alone (n = 15). On day 7 after each procedure, the livers were excised and the number and diameter of the tumor nodules and the cancer index score were determined. Another 90 male BALB/c mice were randomly divided into three groups as described above, and they underwent each procedure (n = 30 each). After each procedure, the livers were excised on days 0, 1, 3, and ICAM-1 and TNF-alpha mRNA expression were examined by real-time RT-PCR using SYBR Green I.The number of tumor nodules and the cancer index score were larger in the CO2 pneumoperitoneum group than in the control group (p0.05). The mean diameter of the tumor nodules was not different among the three groups. The expression of ICAM-1 in the CO2 pneumoperitoneum group was higher than that in the other groups on day 1 (p0.05), and the TNF-alpha mRNA was higher than that in the control group on day 1 (p0.05).CO2 pneumoperitoneum enhances liver metastasis compared with anesthesia alone, and ICAM-1 expression in the liver after the pneumoperitoneum plays an important role in establishing liver metastasis in a murine model.

High-temperature compressive properties at a high strain rate of 103 s−1 were examined in solution treated AZ91 magnesium alloy. Analysis of the flow behavior and microstructural observations indicated that the deformation at high strain rates of ∼103 s−1 proceeds by conventional plastic flow of dislocation glide and twinning even at elevated temperatures.

RNA interference (RNAi) silences gene expression via short interfering 21–23 mer double-stranded RNA (siRNA) segments that guide cognate mRNA degradation in a sequence-specific manner. On the other hand, HIV-1 decoy TAR RNA are known to competitively interact with the HIV-1 Tat protein, to downregulate the enhanced gene expression from the long terminal repeat (LTR) promoters. Here we report that a novel expression construct, encoding both HIV-1 decoy TAR and Vif siRNA, as a single RNA substrate, was expressed under the control of the human U6 promoter, and later the TAR and siRNA were cleaved into their respective separate RNA by the endogenous RNase III-like enzyme. Each of the cleaved HIV-1 anti-genes then synergistically contributed toward enhancing the inhibition efficacy (> 80%) of HIV-1 replication in transduced Jurkat cells. These results suggest that targeting HIV-1 mRNA with simultaneously expressed intracellular decoy TAR and Vif-siRNA could lead to an effective gene therapy strategy for the co...

Background The purpose of this study was to investigate whether the functional liver volume can be measured from changes in expiratory 13 CO 2 levels after intravenous administration of L-[1, 2- 13 C] ornithine, using a rat model of hepatectomy. Materials and methods Under pentobarbital anesthesia, 30%, 70%, or 90% hepatectomy was performed. In the control group, simple laparotomy was performed. Then, 20 mg/kg l -[1, 2- 13 C] ornithine was administered to rats via the femoral vein. A breath test was conducted 20 min after laparotomy. We examined the correlation of the sum of 13 CO 2 output (S) or a single point of 13 CO 2 level (SP) with liver weight/body weight (LW/BW) (%) every 15 min. Results In all of the groups, the ornithine breath test (OBT) graph reached a plateau level at about 6 min. The correlation coefficient between S 15 and LW/BW was highest 0.952 ( P 14 and LW/BW was highest, 0.944 ( P 5–15 and SP 5–15 in all comparisons between groups. Conclusion In the breath test with intravenously administered l -[1, 2- 13 C] ornithine, functional liver volume could be evaluated accurately in a short period.

Liver metastasis is an important prognostic factor in advanced colorectal cancer. Several studies have demonstrated that carbon dioxide (CO2) pneumoperitonem enhances liver metastasis in an animal model. In the present study, we used scanning electron microscopy (SEM) to investigate morphological changes in hepatic vascular endothelium after CO2 pneumoperitoneum in a murine model. Thirty-three male BALB/c mice were randomized to undergo pneumoperitoneum (CO2, air, or helium ), open laparotomy, and anesthesia alone. After each procedure, the animals’ livers were excised at days 0, 1, and 3 and examined by SEM. In the CO2 pneumoperitoneum group, we observed rough surface and derangement of the hepatic vascular endothelial cells and intercellular clefts on day 1. In the other groups, no major morphologic changes were observed at any time. Hepatic vascular endothelium changes after CO2 pneumoperitoneum. Such characteristic changes may play an important role in establishing liver metastasis after CO2 pneumoperitoneum.

Cerebrospinal fluid (CSF) induced neurite retraction of differentiated PC12 cells; the action was observed in 15 min (a rapid response) and the activity further increased until 6 h (a long-acting response) during exposure of CSF to the cells. The CSF action was sensitive to monoglyceride lipase and diminished by homologous desensitization with lysophosphatidic acid (LPA) and by pretreatment with an LPA receptor antagonist Ki16425. Although fresh CSF contains LPA to some extent, the LPA content in the medium was increased during culture of PC12 cells with CSF. The rapid response was mimicked by exogenous LPA, and a long-acting response was duplicated by a recombinant autotaxin, lysophospholipase D (lyso-PLD). Although the lyso-PLD substrate lysophosphatidylcholine (LPC) was not detected in CSF, lyso-PLD activity and an approximately 120-kDa autotaxin protein were detected in CSF. On the other hand, LPC but not lyso-PLD activity was detected in the conditioned medium of a PC12 cell culture without CSF. Among neural cells examined, leptomeningeal cells expressed the highest lyso-PLD activity and autotaxin protein. These results suggest that leptomeningeal cells may work as one of the sources for autotaxin, which may play a critical role in LPA production and thereby regulate axonal and neurite morphological change.

特発性大腸穿孔の中でも稀な横行結腸穿孔の1例を経験した.症例は壊疽性膿皮症で加療中の65歳,女性.平成15年10月22日に出現し経過観察していた腹痛が28日夕方に突然増強し当科紹介となった.腹部所見とCT検査にて,穿孔性腹膜炎と診断し緊急手術となった.横行結腸中央に約3cmの穿孔を認めたため,横行結腸部分切除,腹腔内洗浄ドレナージ,および人工肛門造設を行った.病理組織像では,穿孔部に粘膜と筋層の急峻な断裂像を認めるのみで,潰瘍,腫瘍,憩室などの所見はなかった.特発性横行結腸穿孔は,比較的稀であるが,穿孔性腹膜炎の鑑別疾患の一つとして念頭に置く必要がある.

We investigated the expression of 50 cytochrome P450 (CYP) genes in the post-ischemic reperfused rat liver using a DNA microarray. Thirteen CYPs showed absent expression in all experiments and 2 CYPs were induced by pentobarbital, which was used as an anesthetic. Eight of the remaining 35 CYPs showed significantly decreased expression following ischemia-reperfusion. Monitoring CYP expression may be a powerful approach for elucidation of pathways regulating drug metabolism that may be involved in postischemic reperfusion liver injury. Keywords:: cytochrome P450, postischemic liver reperfusion, gene expression

Background and Aim: Research on gastric lesions developing in response to stress is essential to elucidating the pathogenesis of these lesions as well as the interplay with other factors, including Helicobacter pylori infection and non-steroidal anti-inflammatory drug use. Genes expressed individually or in sets, such as heat shock proteins, growth factors, proto-oncogenes and cyclooxygenases, have been investigated in the stomach. However, gene expression in the stomach after stress exposure have not yet been comprehensively examined. We investigated the gastric gene expression profile in response to stress. Methods: A high-density oligonucleotide array, representing approximately 850 genes, was used to determine gene expression changes in the stomachs of water immersion-restraint stress (WIRS) rats. Results: Fifty-eight genes including expressed sequence tag (EST) genes were upregulated more than twofold in the 30 min-WIRS rat stomach as compared with the control. Concomitantly, five genes were downregulated. Numbers of up- or downregulated genes decreased rapidly at 1 and 2 h of WIRS. Altered gene expression of heat shock proteins, cell cycle regulators, proto-oncogenes and metabolic enzymes were recognized. Several of these genes, including p38 mitogen-activated protein kinase, did not reportedly show gastric expression changes in response to stress. Conclusion: These results suggest that, in addition to the previously identified stress-induced genes, expression of a number of other genes in the stomach is also involved in stress response.

Lettuce big-vein virus (LBVV) is the type species of the genus Varicosavirus and is a two-segmented negative-sense single-stranded RNA virus. The larger LBVV genome segment (RNA1) consists of 6797 nt and encodes an L polymerase that resembles that of rhabdoviruses. Here, the nucleotide sequence of the second LBVV genome segment (RNA2) is reported. LBVV RNA2 consisted of 6081 nt and contained antisense information for five major ORFs: ORF1 (nt 210–1403 on the viral RNA), ORF2 (nt 1493–2494), ORF3 (nt 2617–3489), ORF4 (nt 3843–4337) and ORF5 (nt 4530–5636), which had coding capacities of 44, 36, 32, 19 and 41 kDa, respectively. The gene at the 3′ end of the viral RNA encoded a coat protein, while the other four genes encoded proteins of unknown functions. The 3′-terminal 11 nt of LBVV RNA2 were identical to those of LBVV RNA1, and the 5′-terminal regions of LBVV RNA1 and RNA2 contained a long common nucleotide stretch of about 100 nt. Northern blot analysis using probes specific to the individual ORFs revealed that LBVV transcribes monocistronic RNAs. Analysis of the terminal sequences, and primer extension and RNase H digestion analysis of LBVV mRNAs, suggested that LBVV utilizes a transcription termination/initiation strategy comparable with that of rhabdoviruses.

Dynamic elastic properties and damping properties from room temperature to 673 K were examined in AZ31 magnesium alloy. It was suggested that these properties at elevated temperatures were influenced by the occurrence of grain boundary sliding, which is dependent on the grain size and alloy additions.

The human immunodeficiency virus type-1 (HIV-1)-encoded vif protein is essential for viral replication, virion production, and pathogenicity. HIV-1 vif interacts with the endogenous human APOBEC3G protein (an mRNA editor) in target cells to prevent its virions from encapsidation. Although some studies have established targets within the HIV-1 vif gene that are important for its biologic function, it is however important to further screen for effective therapeutic targets in the vif gene that could interfere with the HIV-1 vif -dependent infectivity and pathogenicity. This report demonstrates that HIV-1 vif antisense RNA fragments constructed within mid-3 ′ region, notably the region spanning nucleic acid positions 5561–5705 (M-3 ′ -AS), significantly inhibited HIV-1 replication in MT-4 and H9-infected cells and reduced the HIV-1 vif mRNA transcripts. These data clearly suggest that the above vif fragment, which corresponds to amino acid residues 96–144, could be an effective novel therapeutic target site for gene therapy applications, for the control and management of HIV-1 infection, due to its strong inhibition of HIV-1 replication in cells.

Background: Accurate staging of lymph node metastasis by sentinel node biopsy is easily achieved in conventional open gastric surgery. Staging is not easily achieved in laparoscopic surgery, however, because of the technical difficulty in identifying sentinel nodes. We developed a laparoscopic method that involves lead shielding for detection of sentinel nodes in gastric cancer and examined the efficacy of this method. Methods: Laparoscopic sentinel node biopsy was performed in 18 patients with early gastric cancer. A combined dye- and radio-guided method was used in the first 10 patients; our radio-guided lead shield method was used in the subsequent eight cases. Laparoscopy-assisted distal gastrectomy was performed in all patients, and dissected nodes were examined by routine hematoxylin and eosin staining. The detection rate was compared between the two groups. Results: The detection rate for sentinel nodes was higher with the radio-guided lead shield method (88%) than with the combined dye- and radio-guided method (40%). Regional lymph node metastasis was recognized in one of 18 patients, and the sentinel node was positive in this case. Conclusions: Use of a lead shield is beneficial for accurate laparoscopic detection of sentinel lymph nodes.

From conventional relative gene expression analyses (Northern blotting, in situ hybridization, and RT-PCR), it has been reported that the expression of control genes, such as glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and β-actin, used as references may be affected by ischemia. Therefore, we extended searching and evaluation at the mRNA level of transcripts whose expression levels were not changed by cerebral ischemia, using a high-density oligonucleotide array and statistical analysis in a rat global cerebral ischemia and reperfusion model. We added a hyperthermic factor and localization factor to ischemia and identified transcripts with a stable expression level under conditions even more disadvantageous than ischemia only. Screening of more than 8,000 transcripts with the Rat Genome U34A array yielded 28 transcripts, which we listed and classified according to their expression level. Widely used control genes, GAPDH and β-actin, were not included, although cyclophilin A was included. In addition, we conducted a functional classification based on gene ontology. Under the functional classification of the 28 transcripts, many genes tended to be associated with metabolism. In conclusion, use of several transcripts is recommended, such as those we identified, as references in the analysis of gene expression in pathological models of ischemia. © 2004 Wiley-Liss, Inc.