Myofibroblasts acquire retinoic acid-producing ability during fibroblast-to-myofibroblast transition following kidney injury.

Tubular injury and interstitial fibrosis are the hallmarks of chronic kidney disease. While recent studies have verified that proximal tubular injury triggers interstitial fibrosis, the impact of fibrosis on tubular injury and regeneration remains poorly understood. We generated a novel mouse model expressing diphtheria toxin receptor on renal fibroblasts to allow for the selective disruption of renal fibroblast function. Administration of diphtheria toxin induced upregulation of the tubular injury marker Ngal and caused tubular proliferation in healthy kidneys, whereas administration of diphtheria toxin attenuated tubular regeneration in fibrotic kidneys. Microarray analysis revealed down-regulation of the retinol biosynthesis pathway in diphtheria toxin-treated kidneys. Healthy proximal tubules expressed retinaldehyde dehydrogenase 2 (RALDH2), a rate-limiting enzyme in retinoic acid biosynthesis. After injury, proximal tubules lost RALDH2 expression, whereas renal fibroblasts acquired strong expression of RALDH2 during the transition to myofibroblasts in several models of kidney injury. The retinoic acid receptor (RAR) RARγ was expressed in proximal tubules both with and without injury, and αB-crystallin, the product of an RAR target gene, was strongly expressed in proximal tubules after injury. Furthermore, BMS493, an inverse agonist of RARs, significantly attenuated tubular proliferation in vitro. In human biopsy tissue from patients with IgA nephropathy, detection of RALDH2 in the interstitium correlated with older age and lower kidney function. These results suggest a role of retinoic acid signaling and cross-talk between fibroblasts and tubular epithelial cells during tubular injury and regeneration, and may suggest a beneficial effect of fibrosis in the early response to injury.

Intercontinental dispersal of HIV-1 subtype B associated with transmission among men who have sex with men in Japan.

UNLABELLED: Transmission clusters of HIV-1 subtype B uniquely associated with the epidemic among men who have sex with men (MSM) in East Asia have recently been identified. Using the Los Alamos HIV sequence database and the UK HIV drug resistance database, we explored possible links between HIV MSM epidemics in East Asia and the rest of the world by using phylogenetic and molecular clock analyses. We found that JP.MSM.B-1, a subtype B MSM variant that accounts for approximately one-third of the infections among Japanese MSM, was detected worldwide, in the United Kingdom (n=13), mainland China (n=3), the United States, Germany, Canada, and Taiwan (n=1 each). Interestingly, 10 United Kingdom samples plus two from Germany and the United States formed a distinct monophyletic subgroup within JP.MSM.B-1. The estimated divergence times of JP.MSM.B-1 and the latter subgroup were ∼1989 and ∼1999, respectively. These dates suggest that JP.MSM.B-1 was circulating for many years in Japan among MSM before disseminating to other countries, most likely through global MSM networks. A significant number of other Asian MSM HIV lineages were also detected in the UK HIV drug resistance database. Our study provides insight into the regional and global dispersal of Asian MSM HIV lineages. Further study of these strains is warranted to elucidate viral migration and the interrelationship of HIV epidemics on a global scale. IMPORTANCE: We previously identified several transmission clusters of HIV-1 subtype B uniquely associated with the epidemic among men who have sex with men (MSM) in East Asia. Using the Los Alamos HIV sequence database and the UK HIV drug resistance database, we explored the possible interplay of HIV MSM epidemics in the different geographic regions and found previously unrecognized interrelationships among the HIV-1 epidemics in East Asia, the United Kingdom, and the rest of the world. Our study provides insight into the regional and global dispersal of Asian MSM HIV lineages and highlights the importance of strengthening HIV monitoring efforts and the need for implementing effective control measures to reduce HIV transmission on a global scale.

[A case report of hepatitis B virus reactivation in a hepatitis B core antibody-positive, hepatitis B surface antigen-negative hemodialysis patient].

Reactivation of the hepatitis B virus (HBV) has been reported in patients receiving immunosuppressive therapy or chemotherapy. We report a case of HBV reactivation in a patient negative for hepatitis B surface antigen (HBsAg), positive for hepatitis B core antibody (anti-HBc), and positive for hepatitis B surface antibody (anti-HBs), who was undergoing chronic maintenance hemodialysis without immunosuppressive therapy or chemotherapy. The patient was an 85-year-old woman with end-stage renal disease due to nephrosclerosis who had undergone maintenance hemodialysis for a year. She had been HBsAg-negative, anti-HBc- and anti-HBs-positive previously, but biannual routine surveillance for HBV showed positivity for HBsAg, negativity for anti-HBs, and positivity for HBV DNA (5.9 log copies/mL). She was asymptomatic, and transaminases were within normal limits. She was dialyzed in an isolated room with a dedicated staff member for the control of infection. HBV is a blood-borne pathogen, which is highly infectious. Hemodialysis is a procedure associated with high risk for blood-borne infection. We should recognize the risk of reactivation of HBV in HBsAg-negative, anti-HBc-positive patients, and consider how to incorporate anti-HBc screening and infection control in isolated anti-HBc-positive hemodialysis patients in clinical practice.

Emergence in Japan of an HIV-1 variant associated with transmission among men who have sex with men (MSM) in China: first indication of the International Dissemination of the Chinese MSM lineage.

A survey of HIV-1 strains circulating in the Tokyo-Kanagawa metropolitan area of Japan during 2004 to 2011 (n = 477) identified six Japanese males (patients 1 to 6), who harbored viruses with genome segments derived from a distinct CRF01_AE variant uniquely found among men who have sex with men (MSM) in China (designated CN.MSM.01-1). These six HIV infections were diagnosed in 2010 and 2011 among MSM (3 of 75) and men with unknown risk factors (3 of 63) and differed from the vast majority of HIV infections among MSM in Japan, which are overwhelmingly characterized by subtype B (239 of 246 [97.2%]). Approximately one-third (91 of 239 [38.1%]) of subtype B strains from MSM in Japan belong to a large monophyletic cluster (designated JP.MSM.B-1). In addition, we identified a smaller subtype B cluster (n = 8) (designated JP.MSM.B-2) that also contains strains from two Chinese MSM living in Japan. Interestingly, patients 5 and 6 were found to be coinfected with CRF01_AE (CN.MSM.01-1) and subtype B (JP.MSM.B-2 or JP.MSM.B-1) variants that are unique to the HIV-1 epidemics among MSM in China and Japan, respectively. Our study demonstrates for the first time the effect of the expanding HIV epidemic among MSM in China on transmission in neighboring countries and shows the ongoing mixing of CRF01_AE and subtype B lineages unique to HIV-1 that cocirculate in MSM populations in East Asia. This finding highlights the importance of strengthening epidemiological surveillance in the region and the need for effective measures to limit transmission among MSM in East Asia.

[Evaluation of a new vesion of the human immunodeficiency virus antigen and antibody combination assay with improved sensitivity in HIV-1 p24 antigen detection].

The performance of a new version of the HIV p24 antigen and antibody combination assays (Genscreen Ultra HIV Ag-Ab) was evaluated by comparing it with three other fourth-generation enzyme immunoassays (Architect HIV Ag/Ab Combo assay, VIDAS HIV DUO Quick and Genscreen Plus HIV Ag-Ab). The assays were examined with 200 HIV positive samples, 1,000 HIV negative samples, 30 samples (28 positives including 24 samples of subtype A, B, B', C, D, F, G, B/D, CRF01_AE in HIV-1 group M, one sample of HIV-1 group O, three samples of HIV-2 and two negatives) of one worldwide HIV performance panel, 59 samples of ten HIV-1 seroconversion panels and the WHO international standard HIV-1 p24 antigen. Both the sensitivity and specificity of Genscreen Ultra HIV Ag-Ab were 100%. All of the 28 positive samples in the worldwide HIV performance panel were positive. The days of the earliest detection in the ten seroconversion panels were the same in three assays (Genscreen Ultra HIV Ag-Ab, Architect HIV Ag/Ab combo assay and VIDAS HIV DUO Quick). Genscreen Plus HIV Ag-Ab which is a former version of the Genscreen Ultra HIV Ag-Ab detected the earliest positive sample one bleed slower than the other three assays in 5 of 10 seroconversion panels. The p24 antigen limit of detection was determined in two ways, using the WHO international standard and three samples from HIV-1 antigen panels; the values obtained were 1IU/mL and 3.5-9.9 pg/mL for Genscreen Ultra HIV Ag-Ab, 1U/mL and 7.1-9.9 pg/mL for Architect HIV Ag/Ab combo assay, 0.5IU/mL and 4.0-7.1 pg/mL for VIDAS HIV DUO Quick, and 32.0-56.5 pg/mL for Genscreen Plus HIV Ag-Ab. In this study, we have shown that Genscreen Ultra HIV Ag-Ab has the sensitivity, specificity and p24 antigen limit of detection that is equal to those of two typical fourth-generation assays. This assay can be considered useful and reliable for HIV screening.

Association of demographics, HCV co-infection, HIV-1 subtypes and genetic clustering with late HIV diagnosis: a retrospective analysis from the Japanese Drug Resistance HIV-1 Surveillance Network.

INTRODUCTION: Late diagnosis of the human immunodeficiency virus (HIV) is a major concern epidemiologically, socially and for national healthcare systems. Although the association of certain demographics with late HIV diagnosis has been reported in several studies, the association of other factors, including clinical and phylogenetic factors, remains unclear. In the present study, we conducted a nationwide analysis to explore the association of demographics, clinical factors, HIV-1 subtypes/circulating recombinant form (CRFs) and genetic clustering with late HIV diagnosis in Japan, where new infections mainly occur among young men who have sex with men (MSM) in urban areas. METHODS: Anonymized data on demographics, clinical factors and HIV genetic sequences from 39.8% of people newly diagnosed with HIV in Japan were collected by the Japanese Drug Resistance HIV-1 Surveillance Network from 2003 to 2019. Factors associated with late HIV diagnosis (defined as HIV diagnosis with a CD4 count <350 cells/µl) were identified using logistic regression. Clusters were identified by HIV-TRACE with a genetic distance threshold of 1.5%. RESULTS: Of the 9422 people newly diagnosed with HIV enrolled in the surveillance network between 2003 and 2019, 7752 individuals with available CD4 count at diagnosis were included. Late HIV diagnosis was observed in 5522 (71.2%) participants. The overall median CD4 count at diagnosis was 221 (IQR: 62-373) cells/µl. Variables independently associated with late HIV diagnosis included age (adjusted odds ratio [aOR] 2.21, 95% CI 1.88-2.59, ≥45 vs. ≤29 years), heterosexual transmission (aOR 1.34, 95% CI 1.11-1.62, vs. MSM), living outside of Tokyo (aOR 1.18, 95% CI 1.05-1.32), hepatitis C virus (HCV) co-infection (aOR 1.42, 95% CI 1.01-1.98) and not belonging to a cluster (aOR 1.30, 95% CI 1.12-1.51). CRF07_BC (aOR 0.34, 95% CI 0.18-0.65, vs. subtype B) was negatively associated with late HIV diagnosis. CONCLUSIONS: In addition to demographic factors, HCV co-infection, HIV-1 subtypes/CRFs and not belonging to a cluster were independently associated with late HIV diagnosis in Japan. These results imply the need for public health programmes aimed at the general population, including but not limited to key populations, to encourage HIV testing.

A long-term survival case of Erdheim-Chester disease on maintenance hemodialysis.

Erdheim-Chester disease, a rare non-Langerhans histiocytosis, involves multiple organs, including kidney. Renal dysfunction sometimes occurs, and is attributed to ureteral obstruction and renal artery stenosis by histiocytic infiltration. However, to our knowledge, case reports of end-stage renal disease requiring renal replacement therapy due to Erdheim-Chester disease are very few. Here, we report a 69-year-old woman who was diagnosed with Erdheim-Chester disease 10 years ago. She had multiple organ involvement, such as bone, skin, heart, pituitary gland, kidney, and retroperitoneum. She had been treated with interferon-alpha, but discontinued after 2 years due to depression and repeated infection. She did not desire treatment with other drugs, so we continued supportive care. Her renal function gradually deteriorated, and hemodialysis was initiated 4 years ago. Subsequently, she is still doing well without any major symptoms. This report describes an unusual case of Erdheim-Chester disease requiring maintenance hemodialysis that longer prognosis than expected was obtained regardless of multiple organ involvement and no specific treatment after interferon-alpha cessation.

CD153/CD30 signaling promotes age-dependent tertiary lymphoid tissue expansion and kidney injury.

Tertiary lymphoid tissues (TLTs) facilitate local T and B cell interactions in chronically inflamed organs. However, the cells and molecular pathways that govern TLT formation are poorly defined. Here, we identified TNF superfamily CD153/CD30 signaling between 2 unique age-dependent lymphocyte subpopulations, CD153+PD-1+CD4+ senescence-associated T (SAT) cells and CD30+T-bet+ age-associated B cells (ABCs), as a driver for TLT expansion. SAT cells, which produced ABC-inducing factors IL-21 and IFN-γ, and ABCs progressively accumulated within TLTs in aged kidneys after injury. Notably, in kidney injury models, CD153 or CD30 deficiency impaired functional SAT cell induction, which resulted in reduced ABC numbers and attenuated TLT formation with improved inflammation, fibrosis, and renal function. Attenuated TLT formation after transplantation of CD153-deficient bone marrow further supported the importance of CD153 in immune cells. Clonal analysis revealed that SAT cells and ABCs in the kidneys arose from both local differentiation and recruitment from the spleen. In the synovium of aged rheumatoid arthritis patients, T peripheral helper/T follicular helper cells and ABCs also expressed CD153 and CD30, respectively. Together, our data reveal a previously unappreciated function of CD153/CD30 signaling in TLT formation and propose targeting the CD153/CD30 signaling pathway as a therapeutic target for slowing kidney disease progression.

Phylodynamic analysis reveals changing transmission dynamics of HIV-1 CRF01_AE in Japan from heterosexuals to men who have sex with men.

BACKGROUND: HIV-1 circulating recombinant form (CRF) 01_AE is the second major subtype in Japan. Our previous study indicated that CRF01_AE was predominantly circulating in heterosexuals/injecting drug users (IDUs). With implications of increased CRF01_AE infections among men who have sex with men (MSM), this study sought to investigate whether the transmission dynamics of CRF01_AE infections in Japan have changed. METHODS: Sequences from 8032 newly diagnosed HIV-1-infected individuals were analysed. For 614 (7.6%) of CRF01_AE cases, clusters were identified and categorised by transmission risks. Median times to the most recent common ancestors (tMRCA) were estimated. RESULTS: The individuals were predominantly Japanese (64%) and male (72%). MSM became the predominant transmission risk from 2014. Thirty transmission clusters (TCs) and 48 pairs, including 40% of individuals, were identified. MSM were approximately five times more likely to be in a TC compared to heterosexuals, and were the major contributors to TCs. tMRCA data suggest that MSM TCs emerged from 1996 and became predominant around 2000. CONCLUSIONS: CRF01_AE has spread among MSM, with frequent and continuous cluster formations, and MSM has become the predominant transmission risk. Our study suggested that CRF01_AE transmission has shifted from heterosexuals/IDUs to MSM. Prevention measures targeting key populations should be considered for controlling CRF01_AE spread.

Hepatitis A virus infection in an HIV-positive man with previously confirmed immunity against hepatitis A virus.

A Japanese man with human immunodeficiency virus (HIV) was detected 9 years ago to have a positive titer for hepatitis A virus (HAV) immunoglobulin (Ig) G, without a history of HAV infection or vaccination. His plasma HIV RNA was well-controlled on antiretroviral therapy for more than 6 years. He developed HAV infection with subsequent reduction of the HAV IgG titer. A decreasing HAV IgG in persons living with HIV might indicate the possibility of HAV reinfection and should prompt the consideration for additional vaccination.

Comparative evaluation of the Geenius HIV 1/2 Confirmatory Assay and the HIV-1 and HIV-2 Western blots in the Japanese population.

Accurate diagnosis of earlier HIV infection is essential for treatment and prevention. Currently, confirmation tests of HIV infection in Japan are performed using Western blot (WB), but WB has several limitations including low sensitivity and cross-reactivity between HIV-1 and HIV-2 antibodies. To address these problems, a new HIV testing algorithm and a more reliable confirmation and HIV-1/2 differentiation assay are required. The Bio-Rad Geenius HIV-1/2 Confirmatory Assay (Geenius) has recently been approved and recommended for use in the revised guidelines for diagnosis of HIV infection by the Center for Disease Control and Prevention (USA). We made comprehensive comparison of the performance of Geenius and the Bio-Rad NEW LAV BLOT 1 and 2 (NLB 1 and 2) which are WB kits for HIV-1 and HIV-2, respectively, to examine if Geenius is a suitable alternative to these WB assays which are now being used in HIV testing in Japan. A total of 166 HIV-1 positive samples (146 from patients with established HIV-1 infection and 20 from patients with acute infection), five HIV-1 seroconversion panels containing 21 samples and 30 HIV-2 positive samples were used. In addition, a total of 140 HIV negative samples containing 10 false-positives on screening tests were examined. The sensitivity of Geenius and NLB 1 for HIV-1 positive samples was 99.3% and 98.6%, respectively. Geenius provided more positive results in the samples from acute infections and detected positivity 0 to 32 days earlier in seroconversion panels than NLB 1. NLB 2 gave positive results in 12.3% of HIV-1 positive samples. The sensitivity of both Geenius and NLB 2 for HIV-2 positive samples was 100%. The specificity of Geenius, NLB 1 and NLB 2 was 98.5%, 81.5% and 90.0%, respectively. Geenius is an attractive alternative to WB for confirmation and differentiation of HIV-1 and HIV-2 infections. The adaptation of Geenius to the HIV testing algorithm may be advantageous for rapid diagnosis and the reduction of testing costs.

Drug-resistant HIV-1 prevalence in patients newly diagnosed with HIV/AIDS in Japan.

The increasing prevalence of drug-resistant HIV transmission has become a critical epidemic in the world today. Studies in developed countries reported 8-27% of newly diagnosed HIV/AIDS patients are infected by drug-resistant strains. To determine the prevalence of drug-resistant HIV-1 among newly diagnosed cases in Japan, eight HIV/AIDS clinical centers, three public health laboratories and the National Institute of Infectious Diseases conducted a nationwide survey. Between January 2003 and December 2004, 575 newly diagnosed HIV/AIDS patients with both acute and chronic infections were enrolled in the study. Twenty-three cases, including three recently infected patients, were infected with HIV-1 having major drug-resistance mutations, including M41L, D67N, L100I, K103N, V106A, M184I, M184V, L210W, and revertant mutations at the 215 codon in reverse transcriptase and M46I in protease encoding regions. In this newly diagnosed population, we also clarified the prevalence of hepatitis virus coinfection, which was 8.8% for HBV and 4.3% for HCV. In conclusion, the drug-resistant transmission rate was 4.0% in Japan. Although this rate is significantly lower than that of other developed countries, this rate almost reaches the threshold at which baseline genotypic resistance testing would be cost-effective for all infected persons before initiating therapy.

[Evaluation of a new real-time PCR assay kit for quantification of human immunodeficiency virus RNA in plasma].

We evaluated the use of COBAS TaqMan HIV-1 for highly pure system (COBAS TaqMan) recently developed as a second-generation quantification of human immunodeficiency virus type 1 (HIV-1) RNA. The linearity, sensitivity, reproducibility, and effects of possibly interfering materials were examined and results compared to those of AMPLICOR HIV-1 MONITOR Test version 1.5 (AMPLICOR) using 6 virus isolates that were all different subtypes. Excellent linearity was obtained at 1.67 x 10(2)-1.73 x 10(6) copies/mL (r2 = 0.991). Sensitivity was 40 copies/mL at a 100% hit rate. Intraexperimental CVs were 27.4-50.8%, and interexperimental CVs were 29.3-81.5%. Although COBAS TaqMan showed an excellent titer correlation with AMPLICOR (r2 = 0.960), mean titer obtained with COBAS TaqMan was 3.1 times higher than that with AMPLICOR (p = 0.002), and 7.1 times higher in a sample of subtype C, suggesting discrepancies in HIV-1 RNA quantification between the two kits. This point should be noted when AMPLICOR is replaced by COBAS TaqMan.

[Evaluation of new fourth-generation human immunodeficiency virus antigen and antibody detection assay with enzyme-linked fluorescent immunoassay].

We evaluated the fourth-generation HIV screening assay VIDAS HIV DUOII (DUOII) based on ELFA for simultaneous detection of anti-HIV-1 and anti-HIV-2 antibodies and HIV-1 p24 antigen through comparison with other HIV antigen-antibody detection assays. Materials were 1228 HIV-negative specimens, 95 HIV-antibody-positive specimens, and HIV commercial panels. The specificity of DUOII was 99.8% and sensitivity 100%, detecting all of HIV-1 group M subtype A, B, B', C, D, A/E, F, G, B/D, HIV-1 group O, and HIV-2. The sensitivity test to HIV-1 p24 antigen was 5pg/ mL, higher than other assays. DUOII was equivalent to or superior in detecting results earlier than other assays in an evaluation using 10 commercial HIV-1 seroconversion panels of primary infection. DUOII detects anti-HIV IgM antibody, so no negative sample was found in the second window between p24 antigen disappearance and raised anti-HIV IgG antibody. DUOII has sufficient specificity and sensitivity for HIV screening, and detects primary infection sooner than other assays. These results indicate that DUOII is useful and reliable in HIV screening.

Analysis of a long-term discrepancy in drug-targeted genes in plasma HIV-1 RNA and PBMC HIV-1 DNA in the same patient.

Drug-resistance genotypes were investigated in a patient under treatment with anti-HIV drugs. Since the drug resistance-associated mutations in plasma HIV-1 RNA and proviral DNA in peripheral blood mononuclear cells (PBMCs) were inconsistent, changes were followed over time, and the discrepancy was shown to persist for a long period. In plasma HIV-1 RNA, D67N, K70R, T215Y, and Y188L were present in the reverse transcriptase (RT) region, and two primary mutations, I84V and L90M, were noted in the protease (Pro) region. In contrast, in proviral DNA, no drug resistance-associated mutations were found in the RT region, and mutations such as L90L/M were only infrequently present in the Pro region. This situation persisted for more than 3 years. In addition, sequencing analysis of the V3 loop in the envelope gene showed that non-syncytium-inducing/macrophage-tropic viruses contribute to acquisition of drug resistance. In this study, drug-resistant viruses were produced primarily at macrophages, and drug-sensitive viruses were maintained in PBMCs as a reservoir.

[Introduction of rapid HIV tests to a public health center in Japan and evaluation of its effects].

OBJECTIVE: As an attempt to make HIV testing more accessible to many people, rapid HIV tests were introduced to a public health center. We evaluated logistic issues and effects of the introduction. METHODS: In January 2003, the Tochigi Prefectural Kennan Public Health and Welfare Center introduced rapid HIV tests simultaneously with conventional HIV tests. We studied the number and types of the clients before and after its introduction as well as comparing them with those of other public centers. Information on rapid HIV test practices was uploaded on the website, "HIV Kensa-Soudan Map" and its effects were also evaluated. RESULTS: The public health center had performed 130 conventional HIV tests in 2002 prior to the introduction of rapid HIV tests but performed 453 HIV tests in 2003 (250% increase). Of those, 404 clients (94%) preferred the rapid tests. Out of the 404 rapid HIV tests performed, five were tested positive but only one was confirmed to be positive (false positive rate 1%). The introduction of rapid HIV tests had a negative impact on other sexually transmitted infection (STI) tests. The access rates to TP antibody tests (rapid tests) and Chlamydia antibody tests (non rapid tests) offered to HIV test recipients declined from 77% to 63% and from 76% to 33%, respectively. However, the actual number of these STI tests increased due to the increased number of HIV tests. Sixty-one per cent of those who came to take the HIV tests referred to the website "HIV Kensa-Soudan Map". During the same time period, the increase rate in the number of those who took the non rapid HIV tests provided at other public health centers in Tochigi prefecture was 0% and the increase rate nationwide was 20%. CONCLUSIONS: The introduction of these tests to public health centers greatly increases access to HIV testing. It was found that the website "HIV Kensa-Soudan Map" was a useful information source for HIV testing. However, because of high false positive rate (about 1%), it is important to establish an effective pre- and post-test counseling system. It is also necessary to prevent the number of tests for other STIs from declining when rapid HIV tests are implemented simultaneously.

Qualitative Real-Time PCR Assay for HIV-1 and HIV-2 RNA.

Because western blotting occasionally causes cross-reactions between human immunodeficiency virus (HIV)-1 and HIV-2, it is difficult to distinguish a coinfection status from a false-positive result. Therefore, we developed a qualitative real-time PCR assay to detect HIV-1 and HIV-2 RNA that can be performed in parallel. Viral RNA extracted from 500 µl of plasma was examined using real-time PCR with minor groove binder probes. Bovine leukemia virus was used as an internal standard. The sensitivity was determined by probit regression analysis using the World Health Organization international standards for HIV-1 and HIV-2. The lower detection limits at a 95% hit rate were 54 IU/ml for HIV-1 and 5.0 IU/ml for HIV-2, which were lower than any HIV-2 assays reported previously. HIV-1 RNA was detected in 51 of 52 HIV-1 seropositive plasma samples. HIV-2 RNA was detected in 7 of 10 HIV-2 seropositive plasma samples. Non-specific signals and cross reactivity between HIV-1 and HIV-2 were not observed in 100 HIV seronegative samples. The assay developed in this study is highly sensitive and specific for the detection of HIV-1 and HIV-2 RNA. The test is expected to be useful for the differential diagnosis of HIV-1 and HIV-2 infections.

Characteristics of Transmitted Drug-Resistant HIV-1 in Recently Infected Treatment-Naive Patients in Japan.

OBJECTIVES: Progress in antiretroviral treatment has led to fewer virological failure cases, but 10%-20% of treatment-naive HIV/AIDS cases are reported to harbor drug-resistant strains, suggesting transmission of drug-resistant HIV. We aimed to determine the trend in prevalence of transmitted drug-resistant (TDR) HIV in Japan, particularly in recently infected patients. METHODS: Drug-resistance test was performed on 3904 HIV-1-infected cases newly diagnosed between 2007 and 2012. The number of cases infected within 6 months [recent seroconverters (RS)] was estimated by BED assay of 2700 plasma samples. Characteristics of RS cases were further analyzed. RESULTS: The overall prevalence of TDR was 9.1%, ranging from 7.3% in 2008% to 12.5% in 2010. Among 1403 subtype B/E/D cases with >50 CD4 T cell counts and >1000 HIV copies per milliliter, 468 (33.4%) were estimated to be RS. The prevalence of RS was significantly higher among cases who were male, Japanese, and men who have sex with men. The prevalence of TDR did not differ significantly between recent and long-term seroconverters (8.5% vs. 9.2%, respectively, P = 0.68). Common mutations in both groups were M46I/L and T215 revertants. Furthermore, sequences with these mutations, K103N and D30N/N88D formed clusters on phylogenetic trees. CONCLUSION: Our study clarified an increase in prevalence of TDR in Japan from 2007 to 2012. The phylogenetic clustering of cases with M46I/L or T215 revertants suggests that HIV with these mutations have become circulating strains. Furthermore, detailed analyses showed that Japanese men who have sex with men are more aware of their risk of HIV infection.

Nurses' Involvement in Patients' Dying and Death: Scale development and validation.

This study reports the development of a measurement scale, The Nurses' Involvement in Patients' Dying and Death Scale (NIPDYDS), which fully captures the experiences of nurses caring for patients' dying and death. Potential items were extracted from narrative data gathered systematically and comprehensively from in-depth interviews with nurses engaged in caring for patients' dying and death. Factor analyses revealed four factors, consisting of 40 total items, with two factors related to the positive aspects of the experience (Deep involvement in facing dying and death and Increased competence in facing dying and death) and two factors related to the negative aspects of the experience (Uncertainty and difficulty dealing with dying and death and Accustomed to dying and death). Validity and reliability of the scale were found to be acceptable. The factorial structure of the NIPDYDS was contrasted to Frommelt's (1991) FATCOD (The Frommelt Attitude Toward Care of the Dying Scale), and the usefulness and limitations of the NIPDYDS were discussed.