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.

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.

Target-cell specificity of fusogenic liposomes: membrane fusion-mediated macromolecule delivery into human blood mononuclear cells.

Fusogenic liposome, a unique vector prepared by fusing ultraviolet-inactivated Sendai virus and liposome, is known to efficiently deliver content into various animal cells through membrane fusion. In this study, we examined the target-cell specificity of fusogenic liposome (FL)-mediated macromolecule delivery into human blood cells using diphtheria toxin fragment A (DTA) as a probe. Among the peripheral blood mononuclear cells (PBMC), FL was able to deliver its encapsulates into CD14+ monocytes and CD4-/CD8- T-cells, but not into CD19+ B-lymphocytes, CD4+ T-cells or CD8+ T-cells. The susceptibility of human leukemia cell lines to FL was similar to that of PBMC; the order of the reactivity was U937 (monoblastic leukemia)>MOLT4, Jurkat (T-lymphoma)>Daudi, BALL1 (B-lymphoma)>K562 (erythroblastic leukemia). Interestingly, FL showed similar binding activity to all of these leukemia cell lines. These findings indicate that, among blood cells, monocytes, monoblastic leukemia cells, CD4-/CD8- T-cells and T-lymphoma cells are preferable targets for FL-mediated macromolecule delivery. This is the first demonstration of the existence of non-permissive cells against FL. Our results also suggest that some molecules on target-cells other than the binding targets of SV-derived protein may participate in fusion between FL and cells.

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.

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.

Increase in the peripheral lymphocyte populations expressing CD54 (ICAM-1) after hyperthermic isolated limb perfusion in patients with malignant melanoma: an analysis of four cases.

The lymphocytes isolated from perfused or non-perfused circulations before, during, and after hyperthermic isolated limb perfusion (HILP) in the four patients with malignant melanoma were analysed for the expression of CD54 (ICAM-1), CD58 (LFA-3), CD4, CD8, HLA class I and class II in order to investigate the mechanism(s) of the activation of such immunocompetent cells as natural killer (NK)-cells or T-lymphocytes by HILP. It was thus found that the lymphocyte populations expressing CD54 increased significantly 1 day after HILP in the four patients examined. The lymphocyte populations expressing CD58 apparently increased. It was also found that the NK-cell and T-lymphocyte activities increased during or after HILP in the present four cases as observed previously in the other melanoma patients. These results indicate that our HILP system may augment the immunological activities through the mechanisms of the induction of CD54 or CD58 expression in the peripheral lymphocytes of the melanoma patients who receive HILP.

High rate of portal thrombosis after splenectomy in patients with esophageal varices and idiopathic portal hypertension.

Data on 126 consecutive patients who were admitted to our clinics from January 1979 to May 1989 were scrutinized to assess changes in portal hemodynamics following splenectomy. Two groups were classified: (1) a group of 106 patients with cirrhosis of the liver and (2) a group of 20 patients with idiopathic portal hypertension (IPH). Portal thrombosis was present in five (25.0%) of the 20 patients with IPH and in two (1.8%) of the 106 patients with cirrhosis of the liver. As seen on celiac arteriography, the mean (+/- SD) diameter of the trunk of the splenic artery and vein was 8.99 +/- 1.55 and 16.2 +/- 3.6 mm, respectively, in patients with IPH, while it was 7.94 +/- 1.28 and 14.2 +/- 3.1 mm, respectively, in patients with cirrhosis of the liver. Changes in portal venous pressure were 78.4 +/- 59.4 mm H2O in patients with IPH and 43.5 +/- 38.7 mm H2O in patients with cirrhosis of the liver. There were no significant differences in the maximum level of thrombocytes in patients with IPH or in those patients with cirrhosis of the liver. These events suggest that portal thrombosis can occur with a significantly higher incidence in patients with IPH than in those patients with cirrhosis of the liver after splenectomy, and a decrease in blood flow in the portal vein may be closely linked to the formation of portal thrombosis after splenectomy in patients with IPH. Preoperative examination of portal hemodynamics must be thorough.

Gene transfer vectors based on Sendai virus.

A gene delivery system is a fundamental technology used in human gene therapy. In order to treat patients suffering from incurable metabolic diseases, we must be able to deliver genes efficiently in situ and induce stable gene expression in non-dividing tissue cells. However, none of the current gene transfer systems (both viral and non-viral) satisfies this goal. In order to develop a novel gene delivery system that is free from the defects of existing gene transfer vectors, we analyzed natural biological phenomena that involve gene transfer and expression, and made artificial components that mimic the functioning of these systems. Our recent results shed light on three major aspects of gene transfer and expression: (1) the direct delivery of DNA into cytoplasm using fusogenic liposomes, (2) the transfer of DNA from cytoplasm to nucleus with a nuclear localization signal, and (3) the stabilization of DNA in the nucleus as an independent replicon. The possible development of a hybrid vector by combining these components is discussed.

HSP70 and c-Fos expression of brain stem hypoglossal nucleus in drowning.

The brain stem hypoglossal nucleus (HN) is the center of nerves innervating the upper respiratory tract and is related to control of mastication, deglutition, speech and respiration. To elucidate the relationship between asphyxia and the HN, we investigated the change of hypoglossal neurons in cases of hanging, strangulation, smothering, choking, drowning and respiratory failure. Using immunohistochemical techniques, we observed the brain stem HN with antibodies against microtubule-associated protein 2 (MAP2), muscarinic acetylcholine receptor (mAChR), c-fos gene product (c-Fos) and 72 kD heat-shock protein (HSP70). MAP2, a cytoskeletal protein of the neuron, is a marker of neuronal damage. Muscarinic AChR was used as a marker of neuronal membrane and ACh signaling. We employed both HSP70 and c-Fos as markers of stress- or damage-related events. We measured the percentage of immunopositive neurons in total neurons of HN. Drowning produced higher expression of HSP70 and c-Fos than other causes of asphyxia, suggesting that drowning induces more severe damage in HN neurons. Furthermore, it was suspected that neuronal changes in drowning might relate to functions of the HN. These observations indicate that immunohistochemical examination of the brain stem HN could provide useful information for determining the cause of asphyxia.

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.

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.