Changes in cerebrovascular disease-related deaths and their location during the COVID-19 pandemic in Japan.

OBJECTIVE: The COVID-19 pandemic placed an enormous strain on healthcare systems and raised concerns for delays in the management of patients with acute cerebrovascular events. In this study, we investigated cerebrovascular excess deaths in Japan. STUDY DESIGN: Vital mortality statistics from January 2012 to May 2022 were obtained from the Japanese Ministry of Health, Labour and Welfare. METHODS: Using quasi-Poisson regression models, we estimated the expected weekly number of cerebrovascular deaths in Japan from January 2020 through May 2022 by place of death. Estimates were calculated for deaths in all locations, as well as for deaths in hospitals, in geriatric health service facilities, and at home. The age subgroups of ≥75 and <75 years were also considered. Weeks with a statistically significant excess of cerebrovascular deaths were determined when the weekly number of observed deaths exceeded the upper bound of 97.5% prediction interval. RESULTS: Excess deaths were noted in June 2021 and became more pronounced from February 2022 onward. The trend was notable among those aged ≥75 years and for those who died in hospitals. With respect to the location of deaths, the excess was significant in geriatric health services facilities from April 2020 to June 2021, whereas no evidence of excess hospital deaths was observed during the same period. CONCLUSIONS: Beginning in the late 2021, excess cerebrovascular deaths coincided with the spread of the Omicron variant and may be associated with increased healthcare burden. In 2020, COVID-19 altered the geography of cerebrovascular deaths, with fewer people dying in hospitals and more dying in geriatric health service facilities and at home.

Excess deaths from COVID-19 in Japan and 47 prefectures from January through June 2021.

OBJECTIVES: In Japan, several studies have reported no excess all-cause deaths (the difference between the observed and expected number of deaths) during the coronavirus disease 2019 (COVID-19) pandemic in 2020. This study aimed to estimate the weekly excess deaths in Japan's 47 prefectures for 2021 until June 27. STUDY DESIGN: Vital statistical data on deaths were obtained from the Ministry of Health, Labour and Welfare of Japan. For this analysis, we used data from January 2012 to June 2021. METHODS: A quasi-Poisson regression was used to estimate the expected weekly number of deaths. Excess deaths were expressed as the range of differences between the observed and expected number of all-cause deaths and the 95% upper bound of the one-sided prediction interval. RESULTS: Since January 2021, excess deaths were observed for the first time in the week corresponding to April 12-18 and have continued through mid-June, with the highest excess percentage occurring in the week corresponding to May 31-June 6 (excess deaths: 1431-2587; excess percentage: 5.95-10.77%). Similarly, excess deaths were observed in consecutive weeks from April to June 2021 in 18 of 47 prefectures. CONCLUSIONS: For the first time since February 2020, when the first COVID-19 death was reported in Japan, excess deaths possibly related to COVID-19 were observed in April 2021 in Japan, during the fourth wave. This may reflect the deaths of non-infected people owing to the disruption that the pandemic has caused.

The relationship between fever rate and telework implementation as a social distancing measure against the COVID-19 pandemic in Japan.

OBJECTIVES: On March 28, the Japanese government decided on the "Basic Policies for Novel Coronavirus Disease Control" and called on the public to thoroughly implement social distancing measures (i.e., behavioral restrictions to limit the frequency and intensity of human contact), especially telework. METHODS: We used population-level questionnaire data from a social networking service (SNS), with 275,560 respondents from March 5 to April 6, to evaluate the relationship between telework implementation and the presence of a fever (body temperature higher than 37.5 °C) within 1 month as a surrogate indicator of COVID-19 infection, by occupation type and age-group. RESULTS: Among company employees, statistical significance was identified in the 15- to 29-year and 30- to 59-year age-groups, showing higher fever rates in the non-teleworker group (for the 15- to 29-year age-group, non-teleworkers: 7.64%; teleworkers: 6.45%; P = 0.02; for the 30- to 59-year age-group, non-teleworkers: 3.46%; teleworkers: 3.14%; P = 0.02). CONCLUSIONS: Telework remains a controversial topic in Japan as the government called for emergency measures. Although caution is warranted in interpreting our findings because our data are limited to the voluntary SNS users, they will be essential to push forward with more measures to promote social distancing measures in the midst of Japan's current tense political climate.

Trend change of the transmission route of COVID-19-related symptoms in Japan.

OBJECTIVES: The Japanese prime minister declared a state of emergency on April 7 2020 to combat the outbreak of coronavirus disease 2019 (COVID-19). This declaration was unique in the sense that it was essentially driven by the voluntary restraint of the residents. We examined the change of the infection route by investigating contact experiences with COVID-19-positive cases. STUDY DESIGN: This study is a population-level questionnaire-based study using a social networking service (SNS). METHODS: To assess the impact of the declaration, this study used population-level questionnaire data collected from an SNS with 121,375 respondents (between March 27 and May 5) to assess the change in transmission routes over the study period, which was measured by investigating the association between COVID-19-related symptoms and (self-reported) contact with COVID-19-infected individuals. RESULTS: The results of this study show that the declaration prevented infections in the workplace, but increased domestic infections as people stayed at home. However, after April 24, workplace infections started to increase again, driven by the increase in community-acquired infections. CONCLUSIONS: While careful interpretation is necessary because our data are self-reported from voluntary SNS users, these findings indicate the impact of the declaration on the change in transmission routes of COVID-19 over time in Japan.

Hepatocyte-specific Bid depletion reduces tumor development by suppressing inflammation-related compensatory proliferation.

Liver cancer is a major health-care concern and its oncogenic mechanisms are still largely unclear. Persistent hepatocyte cell death is a common feature among various chronic liver diseases, the blocking of which presents as logical treatment. Therefore, we aimed at investigating tumor development in mice with hepatocyte-specific Bid depletion--a BH3-only Bcl-2 family member that amplifies apoptotic death signals. Hepatocyte-specific conditional Bid-knockout mice (Bid(Δhep)) were injected with 25 mg/kg diethylnitrosamine (DEN) at 14 days of age, and liver tumorigenesis was investigated 9 months later. Additionally, different models of acute liver injury were used including: acute high-dose DEN challenge, 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) diet and carbon tetrachloride (CCL4) injection. Bid(Δhep) mice developed significantly fewer tumors, showed smaller maximal and average tumor size and reduced tumor incidence. In the acute DEN model, 48 h post injection we observed a significant reduction in liver injury in Bid(Δhep) animals, assessed via serum transaminases and liver histopathology. Furthermore, TNF-α, IL-1ß, cJUN and IL-6 mRNA expression was reduced. These findings were accompanied by reduced compensatory hepatocyte proliferation in Bid(Δhep) mice when compared with controls by immunohistochemistry for Ki67 and proliferating cell nuclear antigen 48 h after DEN injection. In the acute CCL4 model, Bid(Δhep) mice displayed reductions in liver injury and inflammation when compared with controls. No differences in liver injury and serum bilirubin levels were detected in Bid(Δhep) and Bid(flo/flo) mice fed with DDC, which induces bile duct injury and a ductular reaction. Our study demonstrates that in DEN-induced hepatocellular carcinoma, the inhibition of hepatocyte death pathways through Bid deletion protects animals from tumorigenesis. These results suggest that reducing hepatocyte cell death, liver inflammation and compensatory proliferation has a stronger beneficial effect than the potential side effect of enhancing tumor cell survival.

Serial magnetic resonance imaging study of posterior cruciate ligament reconstruction or augmentation using hamstring tendons.

PURPOSE: The purpose of this study was to analyze serial changes in the magnetic resonance imaging (MRI) signals of autograft hamstrings single bundle posterior cruciate ligament (PCL) reconstruction and the effects of remnant preservation (augmentation). MATERIAL AND METHODS: Twenty-two isolated PCL injuries were arthroscopically reconstructed or augmented with hamstring tendons. MRI scans were obtained at 3, 6, and 12 months, and prior to the second-look arthroscopy (average 20.7 months). The patients were divided into 2 groups by remnant preservation: five PCL reconstructions after PCL remnant resection (Group Rec) (23%), and 17 reconstructions preserving the remnant (Group Aug) (77%). The 22 patients were also divided in two groups depending on the location of the PCL tear. There were 9 knees with proximal tear (Type P) (41%) and 13 knees with distal tear (Type D) (59%). The signal intensity and fiber continuity of 4 zones (proximal, middle, distal intra-articular and tibial tunnel zones) were evaluated by the Mariani score. RESULTS: The average MRI evaluation score gradually increased from 6 months through the final MRI. The intra-articular part of the graft exhibited slower maturation (12 months - final scan) as compared with the tibial tunnel (6-12 months). The distal zone underwent better maturation than the proximal or middle zones at all points. In the proximal zone, the score for Group Aug was significantly higher than Group Rec. In the proximal zone, the Type D score with a proximally-preserved remnant was significantly higher than Type P without a proximal remnant. CONCLUSIONS: The hamstring tendons require more than 1 year to achieve low-signal intensity. PCL remnant has a beneficial effect on the maturation of the hamstring graft. LEVEL OF EVIDENCE IV: therapeutic case series.

Proprioceptive function after isolated single-bundle posterior cruciate ligament reconstruction with remnant preservation for chronic posterior cruciate ligament injuries.

INTRODUCTION: Posterior cruciate ligament (PCL) reconstruction using the remnant preserving technique may contribute to improved postoperative posterior stability, graft healing, and proprioception recovery. Although there have been several reports on remnant preserving PCL reconstruction, no study has yet evaluated the proprioceptive functions before and after PCL reconstruction with remnant preservation. The purpose of this study is to retrospectively evaluate the clinical outcomes and proprioceptive function after isolated single-bundle PCL reconstruction with remnant preservation for chronic PCL injuries. HYPOTHESIS: Isolated single-bundle PCL reconstruction with remnant preservation surgery for chronic PCL injuries provides satisfactory clinical outcomes and good recovery of the proprioceptive function. METHODS: Nineteen patients who had undergone isolated single-bundle PCL reconstruction with remnant preservation for chronic PCL injuries were followed up for more than 2 years. The posterior laxity was measured by the gravity sag view, stress radiography and the KT-2000 knee arthrometer. The proprioceptive function was defined as the threshold to detect passive motion (TTDPM). RESULTS: The average Lysholm score significantly improved from 63.7±13.2 preoperatively to 94.4±4.6 at final follow-up. The postoperative posterior laxity significantly improved. Regarding TTDPM, there were no significant differences between the preoperative score and the score at every given time point, regardless of the starting angles and the moving directions of the knees. CONCLUSIONS: The proprioceptive function, defined as TTDPM, is maintained after single-bundle PCL reconstruction with remnant preservation, and the postoperative clinical scores and posterior laxity significantly improve. LEVEL OF EVIDENCE: Level IV, therapeutic case series.

Influence of trunk muscle co-contraction on spinal curvature during sitting cross-legged.

In Asia, many activities of daily living (ADL) are performed while sitting cross-legged on the floor. This sitting posture rotates the pelvis in a more dorsal direction and lumbar lordosis is more flattened than while sitting on a chair. Sitting cross-legged induces a greater load on the intervertebral discs and spine, especially when in a slumped position that is known to increase disc pressure even more and to aggravate chronic low back pain (CLBP). Therefore, it is very important to instruct Asian people about the correct sitting posture. In addition, it is known that co-contraction of the deep spine-stabilizing muscles enhances lumbar segmental stability and the sacroiliac joint. However, little is known about the influence of co-contraction of the trunk deep muscles on spinal curvature while sitting cross-legged on the floor. The purpose of this study was to compare EMG (electromyographic) activity of the trunk muscles while slump cross-legged sitting with that during co-contraction of the trunk muscles and to investigate how this co-contraction influences spinal curvature. Ten healthy male volunteers (21.7 +/- 2.5 years old) without CLBP participated in the study. Bipolar surface electrodes were attached to the rectus abdominis, the obliquus externus abdominis, the obliquus internus abdominis, the lower back extensor muscles (L3) and the multifidus on the right side. EMG signals were continuously recorded while slump sitting cross-legged and during co-contraction of the trunk muscles. They were amplified, band-pass filtered, digitized and stored by a data acquisition system. The average muscle activity values over the five-second sample for each sitting posture were normalized to maximal voluntary contractions (%MVC). While the subjects performed both sitting postures, the curvature of the spine was measured using a skin-surface and hand-held device, the "Spinal Mouse". More significant activities of the trunk muscles, with the exception of the rectus abdominis, were observed during co-contraction of the trunk muscles than while slump sitting cross-legged. The co-contraction of the trunk muscles resulted in significantly less thoracic and lumbar curvature and more sacral angle than while slump sitting cross-legged. The results of this study indicated that co-contraction of the trunk muscles while sitting cross-legged could bring about the correct thoracic and lumbar curvature, and effectively stabilize the lumbopelvic region, and decrease focal stress on passive structures.

Adaptive changes in control of the head while lifting ten times.

We often observe when workers lift objects, they tend to move their heads upward and back. Their movement may be an efficient habit, but little is known about control of the head while lifting. Our primary aim was to investigate the hypothesis that several repetitive liftings would lead to an increase in neck extension. Fifteen healthy male volunteers (20.9 +/- 0.8 years old) were asked to lift a case 10 times without instructions about the starting posture or lifting technique. Motion data and electromyography (EMG) signals were continuously recorded. The lifting time, angles of joints, and distance between the case and the malleolus lateralis at the moment of lifting were calculated from the motion data. EMG signals of the initial 1/10 phase of each lifting time were averaged and normalized to maximal voluntary contractions(% AEMG), and the mean frequencies were calculated by wavelet transform analysis. One-way analysis of variance and post hoc analysis were performed with Dunnett's test (p < 0.05). The cervical extension angle significantly increased, and the distance between the case and the malleolus lateralis significantly decreased for the later lifting times. The % AEMG of the gluteus maximus and the biceps femoris significantly increased for the later lifting times. A moving upward and back of the weight of the head, a decrease in the distance between the body and the case, and increases in tension of the hip extensor muscles could facilitate a righting movement to maintain balance. Therefore, paying attention to head control may be efficient while lifting.

Influence of trunk muscle co-contraction on spinal curvature during sitting reclining against the backrest of a chair.

Today, many office workers frequently adopt a relaxed or slumped sitting posture for many hours, and often people tend to spend their leisure time reclining against the backrest of a chair while sitting for a long time, as when watching television. While sitting, the pelvis rotates backwardly, and lumbar lordosis is flattened. Simultaneously, the load on the intervertebral discs and spine increases. Sitting in a slumped position is known to increase disc pressure even more, and to aggravate chronic low back pain (CLBP). Therefore, it is very important to teach workers and often people about the correct sitting posture. In addition, it has been recognized that co-contraction of the deep spine-stabilizing muscles enhances lumbar segmental stability and the sacro-iliac joint. However, little is known about the influence of co-contraction of the trunk deep muscles on spinal curvature during sitting reclining against the backrest of a chair. The purpose of this study was to compare the EMG (electromyographic) activity of the trunk muscles during slump sitting with that during co-contraction and to investigate how this cocontraction influences spinal curvature. Ten healthy male volunteers (20.8 +/- 0.8 years old) without CLBP participated in the study. Bipolar surface electrodes were attached to the rectus abdominis, the obliquus externus abdominis, the obliquus internus abdominis, the lower back extensor muscles (L3) and the multifidus on the right side. The EMG signals were continuously recorded during slump sitting and co-contraction of the trunk muscles, reclining against the backrest of chair. They were amplified, band-pass filtered, digitized and stored by a data acquisition system. The average muscle activity values over the five-second sample for each sitting posture were normalized to maximal voluntary contractions (%MVC). While the subjects performed both sitting postures, the curvature of the spine was measured using a new skin-surface and hand-held device, the "Spinal Mouse". More significant activities of the trunk muscles, with the exception of the rectus abdominis and the lower back extensor muscles (L3), were observed during co-contraction of the trunk muscles than during slump sitting. The co-contraction of the trunk muscles resulted in significantly less lumbar curvature and more sacral angle than during slump sitting. The thoracic curvature showed no significant change during either sitting posture. The results of this study indicated that co-contraction of the trunk muscles during sitting reclining against the backrest of a chair could bring about the correct lumbar curvature, effectively stabilize the lumbopelvic region, and decrease focal stress on passive structures.

Influences of the position of the head on posture while lifting.

In clinical training of some lower back pain patients, teaching them to control their lumbar lordosis during lifting may be difficult. Therefore, another effective method for lifting technique is required. In standing, head cannot move without some compensating postural adjustment. The purpose of this study was to examine the influence of head position on lifting posture. Fourteen healthy male volunteers (22.6 +/- 4.4 years old) lifted a case while maintaining two different head positions; a downward position and an upright position. In the upright position, activities of the latissimus dorsi and vastus lateralis significantly increased, and these of the biceps femoris significantly decreased during the initial 100 msec phase of lifting. There were no differences in the activities of the upper trapezius, lumbar extensor muscles (L3, L5), and obliquus abdominis under the two conditions. There were also no differences in the lumbar angle when the case was lifted. The flexion angles of the hip, knee, and ankle significantly increased, and the lumbar spine moved closer to the case. Lifting posture was influenced by the head position. Advantages included being able to shift loads on the body from the lower back to the legs, to move the lumbar spine closer to the case, and to relatively increase the moment of lumbar extension. The weight of the head as it moved upward and back, and the weight of the rear part of the body as it moved downward and forward helped to maintain balance.

Influence of trunk muscle co-contraction on spinal curvature during sitting for desk work.

Nowadays, a lot of office workers are forced to sit at a desk for many hours while doing their jobs. While sitting, the pelvis rotates backwardly, and lumbar lordosis is flattened. At the same time, the load on the intervertebral discs and spine increases. Sitting in a slumped position is known to increase disc pressure even more, and to aggravate chronic low back pain (CLBP). Therefore, it is very important to teach workers about the correct sitting posture. In addition, it has been recognized that co-contraction of the deep spine-stabilizing muscles enhances lumbar segmental stability and the sacro-iliac joint. However, little is known about the influence of co-contraction of the trunk deep muscles on spinal curvature during sitting for while doing desk work. The purpose of this study was to compare EMG (electromyographic) activity of the trunk muscles during slump sitting with that during co-contraction of the trunk muscles and to investigate how this co-contraction influences spinal curvature. Ten healthy male volunteers (21.7 +/- 2.5 years old) without CLBP participated in the study. Bipolar surface electrodes were attached to the rectus abdominis, the obliquus externus abdominis, the obliquus internus abdominis, the lower back extensor muscles (L3) and the multifidus on the right side. EMG signals were continuously recorded during slump sitting and co-contraction of the trunk muscles, simulating a desk work sitting posture; i.e., slightly inclined forward. They were amplified, band-pass filtered, digitized and stored by a data acquisition system. The average muscle activity values over the five-second sample for each sitting posture were normalized to maximal voluntary contractions (%MVC). While the subjects performed both sitting postures, the curvature of the spine was measured using a new skin-surface and hand-held device, the "Spinal Mouse". More significant activities of the trunk muscles, with the exception of the rectus abdominis, were observed during co-contraction of the trunk muscles than during slump sitting The co-contraction of the trunk muscles resulted in significantly less lumbar curvature and more sacral angle than during slump sitting. The thoracic curvature showed no significant change during either sitting posture. The results of this study indicated that co-contraction of the trunk muscles during sitting while doing desk work could bring about the correct lumbar curvature, and effectively stabilize the lumbopelvic region, and decrease focal stress on passive structures.

Electromyographic activity of selected trunk muscles during bicycle ergometer exercise and walking.

Recently, active treatment such as exercise has been increasingly advocated for CLBP (chronic low back pain). Specially, exercise to improve fitness has been recommended for the prevention of back injuries. The bicycle ergometer or walking have often been used to improve the fitness of CLBP patients. However, little is known about the activity levels of the trunk muscles during such exercise. In this study, the electromyographic (EMG) activities of the trunk muscles during bicycle ergometer exercises and walking were compared and the load level on these muscles during such exercises was investigated. The present study provides basic information concerning fitness exercise in CLBP patients. Eleven healthy male volunteers (21.7 +/- 2.5 years old) without low back pain participated in the study. Bipolar surface electrodes were attached to the right side of the rectus abdominis, the obliquus externus abdominis and lower back extensor muscles (L3). EMG signals were continuously recorded while walking and during gradual loading exercises and normalized to maximal voluntary contractions (% MVC). One way analysis of variance (ANOVA) was performed on the % MVC from each exercise and walking for each of the three trunk muscle sites (p < 0.05). The rectus abdominis muscle showed activity of about 6% MVC during any grade of exercise and walking and no significant differences were found between these forms of exercise. The obliquus externus abdominis muscle showed about 30% MVC during any grade of exercise and walking, but no significant difference was found between them. The low back muscles showed activity of about 12% MVC while walking, whereas activity level increased as the exercise load using the bicycle ergometer increased. More significant low back muscles activity was observed while walking than during exercises of 25 w and 50 w. The results of this study indicated that exercise using the bicycle ergometer should be useful for maintaining or improving fitness in CLBP patients, because it results in less load on the trunk muscles and relatively more oxygen uptake than walking.

Effect of static stretch on fatigue of lumbar muscles induced by prolonged contraction.

Low back pain (LBP) is believed to be the result of fatigue of the back extensor muscles induced by prolonged contraction. Although static stretch has been considered to promote recovery from such muscle fatigue by relaxation, little is known about the effect of stretch on muscle fatigue, especially in the back extensor muscles. The purpose of this study was to investigate the effect of static stretch on prolonged contraction-inducedfatigue of the back extensor muscles using electromyographic (EMG) spectral analysis in ten healthy volunteers. Bipolar surface electrodes were placed on the longissimus and the multifidus muscles. EMG signals were collected during two trials of a Sorensen trunk-holding test for two minutes, with a five-minute rest period between the two trials (control test). All subjects were asked to perform the same trials, with one-minute of static stretch and a four-minute rest period (stretch test). Maintaining the knees toward the chest in supine position resulted in static stretch. The median frequency (MF) was calculated using a spectrum analysis program, and the MF slope over time was computed by linear regression analysis, and normalized with the y-intercept. The decreasing rate of the normalized MF slope between two trials in two tests was compared. The decreasing rate of the normalized MF slope of the second trial was larger than that of the first trial in the control test (p < 0.05), but no significant difference was observed in the stretch test. The results indicated that static stretch had a significant effect on the recovery from fatigue of the back extensor muscles, since it influenced the decreasing rate of the normalized MF slope.

Protein transduction domain of HIV-1 Tat protein promotes efficient delivery of DNA into mammalian cells.

The plasma membrane of mammalian cells is one of the tight barriers against gene transfer by synthetic delivery systems. Various agents have been used to facilitate gene transfer by destabilizing the endosomal membrane under acidic conditions, but their utility is limited, especially for gene transfer in vivo. In this article, we report that the protein transduction domain of human immunodeficiency virus type 1 Tat protein (Tat peptide) greatly facilitates gene transfer via membrane destabilization. We constructed recombinant lambda phage particles displaying Tat peptide on their surfaces and carrying mammalian marker genes as part of their genomes (Tat-phage). We demonstrate that, when animal cells are briefly exposed to Tat-phage, significant expression of phage marker genes is induced with no harmful effects to the cells. In contrast, recombinant phage displaying other functional peptides, such as the integrin-binding domain or a nuclear localization signal, could not induce detectable marker gene expression. The expression of marker genes induced by Tat-phage is not affected by endosomotropic agents but is partially impaired by inhibitors of caveolae formation. These data suggest that Tat peptide will become a useful component of synthetic delivery vehicles that promote gene transfer independently of the classical endocytic pathway.

Nuclear targeting of DNA.

The nuclear membrane is a tight barrier for cytoplasmic proteins, but nuclear proteins have the intrinsic ability to overcome this barrier by an active signal-mediated process. Specific cytoplasmic carrier proteins have the responsibility to escort these proteins into the nucleus through the nuclear pore. The nuclear membrane is also a tight barrier for exogenous DNA delivered by synthetic vehicles, while many of the karyophilic viruses have a mechanism to actively deliver their genome through the nuclear pore. Virus DNA and RNA cannot move into the nucleus by themselves and require the viral structural proteins for efficient nuclear transport. In this article, we review the recent progress in understanding the mechanism of the nuclear transport of proteins and the virus genome, and discuss the possibility of developing synthetic gene-delivery systems based on these outcomes.

A novel strategy for cancer therapy by mutated mammalian degenerin gene transfer.

Mammalian degenerin (MDEG) is a member of the amiloride-sensitive sodium ion channel family, and its site-directed active mutant (MDEG-G430F) induces massive Na+ influx into cells, leading to cell ballooning and cell bursting. We attempted a novel therapeutic approach for gastric cancers by transferring MDEG-G430F into cancer cells using tumor-specific promoters. In carcinoembryonic antigen (CEA)-producing gastric cancer cells, the level of cell death observed when MDEG-G430F was used with a CEA promoter was similar to that observed when using a potent nonspecific promoter such as the cytomegalovirus promoter. In an in vivo study, fusogenic liposome complexes containing MDEG-G430F driven by the CEA promoter were injected intraperitoneally into CEA-producing gastric cancer cells in a mouse peritoneal dissemination model. Although all 15 of the control mice were dead by 50 days postinoculation, 13 of the 15 mice treated with MDEG-G430F survived. These results indicate that transferring MDEG-G430F into cancer tissues using tumor-specific promoters can achieve striking and selective cancer cell death irrespective of the transcriptional efficiency of the promoters used in vivo, and suggest that this approach is a promising new strategy for cancer gene therapy.

TRF1 is a critical trans-acting factor required for de novo telomere formation in human cells.

The duplex telomere repeat (TTAGGG)(n) is an essential cis-acting element of the mammalian telomere, and an exogenous telomere repeat can induce chromosome breakage and de novo telomere formation at the site of a break (telomere seeding). Telomere seeding requires the telomere repeat (TTAGGG)(n) more stringently than does an in vitro telomerase assay, suggesting that it reflects the activity of a critical trans-acting element of the functional telomere, in addition to telomerase. Furthermore, telomere seeding is induced at a frequency fluctuating widely among human cell lines, suggesting variation in the activity of this hypothetical factor among cells. In this study, we investigated the cellular factor(s) required for telomere formation using the frequency of telomere seeding as an index and identified TRF1, one of the telomere repeat binding proteins, as an essential trans-acting factor. The exogenous telomere repeat induces telomere formation at a frequency determined by the availability of TRF1, even in telomerase-negative cells. Our study shows clearly that TRF1 has a novel physiological significance distinct from its role as a regulator of telomere length in the endogenous chromosome. The possible role of TRF1 in cell aging and immortalization is discussed.

Effect of toluene inhalation on astrocytes and neurotrophic factor in rat brain.

Toluene, an abused substance in Japan, is a neurotoxic chemical that has been shown to have neurobehavioral and electrophysiological effects. In previous work, both acute and chronic effects of toluene on cells have been studied extensively. However, although glial cells are thought to play an important role in the survival of neurons in the brain, the effect of toluene on glial cell function has not yet been characterized. To elucidate this, the effect of toluene inhalation on astrocytes in rat brain was examined. Toluene exposure (1500 ppm for 4 h on 4-10 days) augmented glial fibrillary acidic protein (GFAP) immunoreactivity, particularly in the hippocampus and cerebellum. Quantitative analysis showed that toluene inhalation markedly enhanced GFAP expression in the hippocampus and cerebellum. In both regions, proliferating cell nuclear antigen (PCNA) showed no obvious changes, but glutamine synthetase (GS)-immunoreactive cells were markedly increased by toluene exposure. Thus, the elevation of GFAP expression was induced by astrocyte activation rather than by cell proliferation. If toluene exposure activates astrocytes, astrocytes may play a role in the neurophysiological changes observed in toluene intoxication. A neurotrophic factor, basic fibroblast growth factor (b-FGF) was observed immunohistochemically in the capillary vessel walls in the hippocampus and the cerebellum of toluene-intoxicated rats. Basic-FGF may have induced GFAP expression both in the hippocampus and the cerebellum. So, other neurotrophic factors may affect the difference of GFAP elevation between the hippocampus and the cerebellum. These differences may relate to neurobehavioral function of each brain part after toluene exposure.