The Tol2 transposon system mediates the genetic engineering of T-cells with CD19-specific chimeric antigen receptors for B-cell malignancies.

Engineered T-cell therapy using a CD19-specific chimeric antigen receptor (CD19-CAR) is a promising strategy for the treatment of advanced B-cell malignancies. Gene transfer of CARs to T-cells has widely relied on retroviral vectors, but transposon-based gene transfer has recently emerged as a suitable nonviral method to mediate stable transgene expression. The advantages of transposon vectors compared with viral vectors include their simplicity and cost-effectiveness. We used the Tol2 transposon system to stably transfer CD19-CAR into human T-cells. Normal human peripheral blood lymphocytes were co-nucleofected with the Tol2 transposon donor plasmid carrying CD19-CAR and the transposase expression plasmid and were selectively propagated on NIH3T3 cells expressing human CD19. Expanded CD3(+) T-cells with stable and high-level transgene expression (~95%) produced interferon-γ upon stimulation with CD19 and specifically lysed Raji cells, a CD19(+) human B-cell lymphoma cell line. Adoptive transfer of these T-cells suppressed tumor progression in Raji tumor-bearing Rag2(-/-)γc(-/-) immunodeficient mice compared with control mice. These results demonstrate that the Tol2 transposon system could be used to express CD19-CAR in genetically engineered T-cells for the treatment of refractory B-cell malignancies.

Electromyogram biofeedback training for daytime clenching and its effect on sleep bruxism.

Bruxism contributes to the development of temporomandibular disorders as well as causes dental problems. Although it is an important issue in clinical dentistry, no treatment approaches have been proven effective. This study aimed to use electromyogram (EMG) biofeedback (BF) training to improve awake bruxism (AB) and examine its effect on sleep bruxism (SB). Twelve male participants (mean age, 26·8 ± 2·5 years) with subjective symptoms of AB or a diagnosis of SB were randomly divided into BF (n = 7) and control (CO, n = 5) groups to undergo 5-h daytime and night-time EMG measurements for three consecutive weeks. EMG electrodes were placed over the temporalis muscle on the habitual masticatory side. Those in the BF group underwent BF training to remind them of the occurrence of undesirable clenching activity when excessive EMG activity of certain burst duration was generated in week 2. Then, EMGs were recorded at week 3 as the post-BF test. Those in the CO group underwent EMG measurement without any EMG BF training throughout the study period. Although the number of tonic EMG events did not show statistically significant differences among weeks 1-3 in the CO group, events in weeks 2 and 3 decreased significantly compared with those in week 1, both daytime and night-time, in the BF group (P < 0·05, Scheffé's test). This study results suggest that EMG BF to improve AB tonic EMG events can also provide an effective approach to regulate SB tonic EMG events.

Signature current of SO2-induced bronchitis in rabbit.

To investigate abnormalities of airway epithelial ion transport underlying chronic inflammatory airway diseases, we performed electrophysiological, histological, and molecular biological experiments using rabbits exposed to SO2 as a model of bronchitis. By comparison with control, the SO2-exposed trachea exhibited decreased short circuit current (Isc) and conductance associated with increased potential difference. In normal trachea, apical ATP induced a transient Isc activation followed by a suppression, whereas the bronchitis model exhibited a prolonged activation without suppression. This pathological ATP response was abolished by diphenylamine 2-carboxylate or Cl--free bath solution. A significant increase in net Cl- flux toward the lumen was observed after ATP in our bronchitis model. Isoproterenol or adenosine evoked a sustained Isc increase in SO2-exposed, but not in normal, tracheas. The Northern blot analysis showed a strong expression of cystic fibrosis transmembrane conductance regulator (CFTR) mRNA in SO2-exposed epithelium. The immunohistochemical study revealed a positive label of CFTR on cells located luminally only in SO2-exposed rabbits. We concluded that the prolonged ATP response in our bronchitis model was of a superimposed normal and adenosine-activated current. The latter current was also activated by isoproterenol and appeared as a signature current for the bronchitis model airway. This was likely mediated by CFTR expressed in the course of chronic inflammation.

Assessing age-related change in Japanese mental foramen opening direction using multidetector computed tomography.

OBJECTIVE: The purpose of this study was to investigate how the opening direction of the mental foramen (MF) changes with age in a Japanese population using multi-detector computed tomography (MDCT). METHODS: Post-mortem MDCT scans of 121 Japanese subjects (66 males and 55 females) were carried out where all subjects possessed at least twenty teeth, including molar teeth, in the upper and lower jaws. Two angles of the mental foramen opening were measured, namely the superior-inferior angle in the coronal plane and anterior-posterior angle in the transverse plane, on the CT reconstructed images. The associations between age and these two angles were evaluated using a multiple regression analysis. RESULTS: For male subjects, the relationship between the superior-inferior angle and age was a quadratic curve (p < 0.001). This angle increased until the subject reached their early 50s and then the angle decreased with age. In the transverse plane, there was a linear relation between the anterior-posterior angle and age (p=0.002).It was noted also that the angle decreased with age. By contrast, however, no significant associations between the two angles and age for either measurement were noted for female subjects. This study demonstrated that the opening direction of the mental foramen changes with age in Japanese male subjects. By contrast this change in the opening direction of the mental foramen was not demonstrated in Japanese female subjects. In male subjects, the opening direction moves superiorly until the individual reaches their early 50s, and then moves inferiorly with advancing age. It also shifts from a posterior to an anterior position with age. CONCLUSION: These observed change differ from the results of previous studies. The findings could be useful for forensic science as they demonstrate a change in the position of mental foramen in a sample of contemporaneous male Japanese subjects.

Effects of bronchoconstriction and external resistive loading on the sensation of dyspnea.

To determine whether the intensity of dyspnea at a given level of respiratory motor output differs between bronchoconstriction and the presence of an external resistance, we compared the sensation of difficulty in breathing during isocapnic voluntary hyperventilation in six normal subjects. An external resistance of 1.9 cmH2O.1-1.s was applied during both inspiration and expiration. To induce bronchoconstriction, histamine aerosol (5 mg/ml) was inhaled until airway resistance (Raw) increased to a level approximately equal to the subject's control Raw plus the added external resistance. To clarify the role of vagal afferents on the genesis of dyspnea during both forms of obstruction to airflow, the effect of airway anesthesia by lidocaine aerosol inhalation was also examined after histamine and during external resistive loading. The sensation of difficulty in breathing was rated at 30-s intervals on a visual analog scale during isocapnic voluntary hyperpnea, in which the subjects were asked to copy an oscilloscope volume trace obtained previously during progressive hypercapnia. Histamine inhalation significantly increased the intensity of the dyspneic sensation over the equivalent external resistive load at the same levels of ventilation and occlusion pressure during voluntary hyperpnea. Inhaled lidocaine decreased the sensation of dyspnea during bronchoconstriction with no change in Raw, but it did not significantly change the sensation during external resistive loading. These results suggest that afferent vagal activity plays a role in the genesis of dyspnea during bronchoconstriction.

Effects of pharyngeal lubrication on the opening of obstructed upper airway.

We examined the effect of electrical stimulation of the hypoglossal nerve and pharyngeal lubrication with artificial surfactant (Surfactant T-A) on the opening of obstructed upper airway in nine anesthetized supine dogs. The upper airway was isolated from the lower airway by transecting the cervical trachea. Upper airway obstruction was induced by applying constant negative pressures (5, 10, 20, and 30 cmH2O) on the rostral cut end of the trachea. Peripheral cut ends of the hypoglossal nerves were electrically stimulated by square-wave pulses at various frequencies from 10 to 30 Hz (0.2-ms duration, 5-7 V), and the critical stimulating frequency necessary for opening the obstructed upper airway was measured at each driving pressure before and after pharyngeal lubrication with artificial surfactant. The critical stimulation frequency for upper airway opening significantly increased as upper airway pressure became more negative and significantly decreased with lubrication of the upper airway. These findings suggest that greater muscle tone of the genioglossus is needed to open the occluded upper airway with larger negative intraluminal pressure and that lubrication of the pharyngeal mucosa with artificial surfactant facilitates reopening of the upper airway.

Rectal submucosal tumor-like lesion originating from intestinal tuberculosis.

We present the case of a 55-year-old man who underwent transsacral local excision for a rectal submucosal tumor-like lesion suspected to originate from tuberculosis. The lesion, 2 cm in size, was found incidentally in the posterior wall of the lower rectum during anal fistulectomy. The lesion was apart from the primary crypt of the anal fistula. Barium enema and colonoscopy revealed a protuberant submucosal growth with a shallow depression of the overlying mucosa. Although computed tomography and magnetic resonance imaging showed a well defined round mass within the rectal wall, digital rectal examination suggested extramural origin. Since repeated endoscopic biopsies were negative, we selected the transsacral approach for excisional biopsy to achieve histological diagnosis. The lesion was confined to the rectal wall and the full-thickness rectal wall was excised. Histologically, a foreign-body granuloma with acute inflammation was the main component of the lesion. Caseating granulomas and Langhans' giant cells, consistent with tuberculosis, were also found.

Transmitter biochemistry and histochemistry of the hypoplastic cerebellum in mice after neonatal administration of cytosine arabinoside.

Administration of cytosine arabinoside, a DNA polymerase inhibitor, to mice (50 mg/kg body weight) on postnatal days 2, 3 and 4 resulted in severe hypoplasia of the cerebellar cortex accompanied by a large decrease in the cerebellar DNA and RNA contents. On postnatal days 10, 20, 30 and 70, concentrations of the putative neurotransmitters, noradrenaline and serotonin, were significantly increased in the affected cerebellum. In contrast, gamma-aminobutyric acid concentration was little changed, although a decrease in the content of gamma-aminobutyric acid per cerebellum was observed from postnatal day 20. No changes were found in the concentrations of these transmitter candidates nor in dopamine in the cerebral cortex or the brain stem. Histofluorescence observations revealed abnormally dense noradrenergic fibers in a poorly organized cerebellar cortex. The distribution of these fibers was characterized by a honeycomb pattern in the Purkinje cell and granular layers, and a dense plexus in the lower part of the thin molecular layer. However, the significance of neither the density of the noradrenergic fibers nor their abnormal distribution was apparent from the results of this study.

Are lower legal blood alcohol limits and a combination of sanctions desirable in reducing drunken driver-involved traffic fatalities and traffic accidents?

In 1970 and 1978, a set of strict new countermeasures against drunk driving went in to effect in Japan. Analysis of official statistics of motor vehicle fatality data have indicated that alcohol involvement in fatal crashes has declined substantially in Japan since 1970. From the beginning of 1970 to the late 1980s and 1990s, public awareness of and tolerance for the problem of alcohol-impaired driving changed dramatically, as shown in this study. Further it seems that attitudes in Japan on drink driving have improved over the last 20 years or so, instep with a major program of government action. As well as being part of a long running campaign to reduce alcohol related road deaths and injuries, these accident savings are an important part of a national strategy which began in 1970, comprising a well structured legislative program introducing a lower legal limit, progressive penalties for those above the legal limit, over and above Police enforcement strategies underpinning the law and reinforcing the publicity massage. Enactment of the lower legal blood alcohol level with a combination of other severe sanctions is desirable for prevention of alcohol-related traffic casualties, DWI, and accidents, which is shown in this study. Finally, much of the current reduction in alcohol-related fatalities and morbidity reflects that Japanese society has largely endorsed alcohol impaired driving as a socially undesirable behavior. However, this study suggests that it is necessary for policy makers to understand that the DUI problem in Japan must be handled with diverse approaches, rather than relying exclusively on the deterrence based laws.

Spontaneous osteoarthritic lesions in a new mutant strain of the mouse.

In our laboratory, mice showing signs of osteoarthritic lesions with cinnamon colored (yellowish-brown) hair were discovered in a colony of B6C3F1 mice. This mouse is characterized by tiptoe walking and swelling and ankylotic changes in the ankle joint. As to radiographic findings, osteoarthritic changes, such as erosion and/or fusion of the bone tissue, were evident in the ankle joints. Histopathological characteristics included irregularity of articular surfaces caused by fissuring and/or erosion with degeneration of articular cartilage, as well as osteophytes with abnormal proliferation of chondrocytes in joint margin regions. Subsequently, ankylotic changes in the ankle joints were completed in the formation of a cartilaginous bridge and fusion of articular cavity with abnormal proliferation of cartilaginous or bone tissues. This mouse strain may provide an additional animal model that is valuable in the study of human osteoarthritis (OA).

Lichen planus with involvement of all twenty nails and the oral mucous membrane.

A 57-year-old man had had deformities of all ten fingernails for one and a half years before presentation and deformities of all ten toenails for the previous six months. The surfaces of the nails were rough, with excessive longitudinal striations. The bases of the nails were slightly hypertrophic, and the tips were atrophic and itchy. A longitudinal nail biopsy including the nail matrix revealed the typical histology of lichen planus. Reticulated pigmentation, maceration, and erosion on the buccal mucous membrane were also discovered. Histological analysis of the buccal mucous membrane revealed lichen planus intermingled with eosinophils. Immunological blood analysis revealed elevated CD4+ T cells and CD4/CD8 ratio. He worked as a tinsmith and had dental metal. The metal series patch test revealed positive reactions to chromate and tin. Treatment with systemic steroids was quite effective in treating the nail lesions.

Effect of propranolol on baroreflex control of heart rate in young patients with essential hypertension.

The effect of propranolol on baroreflex control of heart rate was studied in 12 young patients with essential hypertension and was compared with that of 12 age-matched normotensive subjects. Resting heart rate and cardiac index in patients with essential hypertension were significantly higher than those of control subjects (p less than 0.001, p less than 0.01, respectively). The reduction of heart rate and cardiac index after propranolol were significantly greater in patients with essential hypertension than in control subjects (p less than 0.01, p less than 0.05, respectively). In the control state, the baroreflex slope in young patients with essential hypertension was significantly less than that of control subjects (p less than 0.01). After propranolol, the baroreflex slope was significantly increased in control subjects (p less than 0.01), but remained unchanged in patients with essential hypertension. These results suggest, first that the antihypertensive action of propranolol must be due to a mechanism other than the augmented baroreflex function and, second that increased beta-adrenergic activity is not a factor contributing to impaired baroreflex function in young patients with essential hypertension.

Histopathological characterization of spontaneously developing osteoarthropathy in the BCBC/Y mouse established newly from B6C3F1 mice.

A new breed of mouse showing cinnamon-color of hair and osteoarthropathy was born from mouse of the B6C3F1 strain, and was named the BCBC/Y mouse. The incidence and severity of the articular lesions increased with aging, and by 10 to 22 months of age, many mice showed moderate-severe motor paresis with ankylotic changes of the limb joints. Abnormal radiographic changes were observed at many joints with increasing age. There was bone and joint deformity, followed by progressive osteoarthritic changes and ankylosis of the limb joints. Histopathologically, degenerative changes, followed by loosening, fissuring and erosion of the articular cartilage were observed at early stage of articular lesions. Unilateral progressive changes, characterized by joint fusion with abnormal osteophytes, were observed at late stage of articular lesions with aging. For successive breeding to establish a homogeneous strain, generation mice retained the symptoms of the primary mice. Thus, these BCBC/Y mice may be a useful as mode to elucidate the hereditary background and hyperplastic changes of the osteochondrocytes for pathogenesis of the osteoarthropathy.

[A case report of multiple pseudoaneurysms of the left ventricle following acute myocardial infarction].

Multiple pseudoaneurysms of the left ventricle following acute myocardial infarction were successfully repaired in a 50-year-old man. Three days after onset of acute myocardial infarction, we electively performed PTCA on the right coronary artery which was completely occluded at the proximal segment. One month after PTCA, the left ventriculogram revealed multiple pseudoaneurysms of the left ventricle. Primary closure of orifices of the pseudoaneurysms was performed using pledgeted mattress sutures under cardiopulmonary bypass. Histology of aneurysm confirmed the diagnosis of pseudoaneurysm.

[Risk factors for contrast nephropathy in diabetic patients undergoing cardioangiography].

Risk factors for contrast nephropathy were prospectively studied in 17 patients with non-insulin dependent diabetes mellitus undergoing cardioangiography. Contrast nephropathy, defined as a serum creatinine increase of greater than 25% at 3 day after angiography, occurred in 29.4% of diabetic patients. Patients who developed contrast nephropathy had significantly higher serum creatinine (Cr), fractional excretion of sodium (FENa), urinary albumin excretion rate (AER), and lower 24hr Ccr than patients who did not (Cr: 1.5 +/- 0.3 mg/dl vs. 0.8 +/- 0.1 mg/dl, FENa: 1.9 +/- 0.5% vs. 0.6 +/- 0.1%, AER: 522 +/- 335 micrograms/min vs. 27 +/- 13 micrograms/min, 24hr Ccr: 39.1 +/- 11.6 ml/min vs. 86.2 +/- 9.3 ml/min, P < 0.05). Contrast nephropathy developed in all of two patients with overt proteinuria (AER more than 200 micrograms/min), but none of eight patients with normoalbuminuria (AER below 15 micrograms/min). Three of seven patients with microalbuminuria developed contrast nephropathy, and two of them had advanced nephropathy. FENa obtained next day was significantly elevated over baseline in patients with contrast nephropathy (1.9 +/- 0.5% vs. 9.7 +/- 4.5%, P < 0.05), but unchanged in patients without contrast nephropathy. The rise in C beta 2-microglobulin/Ccr and enzymuria was noted in both group. Percentage decrease of Ccr on the next day was positively correlated with FENa before angiography (r = 0.645, p < 0.01). Of 24hr Ccr, AER, and FENa before angiography, FENa was revealed as a statistically significant discriminant factor for contrast nephropathy by stepwise discriminant analysis (p = 0.0008). These results suggest that contrast nephropathy develops predominantly in the stage not of incipient but of overt diabetic nephropathy indicated by a decline of glomerular filtration, overt proteinuria, and tubular dysfunction. Of them, tubular dysfunction may be the most important risk factor for contrast nephropathy.

Abnormal baroreflex control of heart rate in normotensive young subjects with a family history of essential hypertension.

This study was performed to determine whether baroreflex control of heart rate is impaired in young normotensive subjects with hypertensive relatives. Blood pressure, baroreflex function, pressor response to phenylephrine, aortic distensibility (pulse pressure/stroke volume) and minimal forearm vascular resistance (minFVR) were assessed in 13 normotensive subjects with hypertensive relatives (group A) and 11 normotensive subjects with no family history of hypertension (group B). Baroreflex sensitivity was significantly lower in group A than in group B (9.4 +/- 1.7 versus 17.5 +/- 1.7 ms/mmHg, P less than 0.01), although blood pressure, aortic distensibility and minFVR were not different between two groups. Pressor response to phenylephrine was identical for both groups. These results suggest that baroreflex control of heart rate is impaired in normotensive young subjects with hypertensive relatives and this defect may be inherited rather than the result of elevated blood pressure and decrease aortic distensibility.

Impact of the 1970 legal BAC 0.05 mg% limit legislation on drunk-driver-involved traffic fatalities, accidents, and DWI in Japan.

Official statistics of motor vehicle fatality data have indicated that alcohol involvement in fatal crashes has declined substantially in Japan since 1970. The national campaign against drunken driving in Japan provides a natural experiment in which to test the predictions of deterrence theory. Utilizing official data over the 1960-1995 period, we report conclusive evidence that 1970s legislation is having a measurable and long-term effect on alcohol-related motor vehicle fatalities. Enactment of the lower legal blood alcohol limit with a combination of other severe sanctions has been more desirable in the prevention of alcohol-related fatalities in the long term as shown in this study. Further extensive research is necessary to investigate performance at the lower legal limit in other societies in order to inspire a strong "international lobby" to support the reduced legal blood alcohol limit for drivers all over the world, as there is no doubt that a reduction in alcohol-impaired driving will result in a substantial savings of human lives and resource worldwide.

Effects of flutropium on experimental models of drug- and allergy-induced rhinitis in guinea pigs.

The effects of flutropium on histamine (Hist)-induced increase in intranasal pressure in non-sensitized guinea pigs and nasal mucosa capillary permeability in passively sensitized guinea pigs were investigated. Flutropium (0.3%), atropine (0.3%), diphenhydramine (0.01%) and cimetidine (0.1%) were directly inhaled into the nasal cavities by an ultrasonic nebulizer for 20 min, followed by inhalation of Hist (0.1%) for 10 min. Flutropium, atropine and diphenhydramine had an inhibitory action on the Hist-induced increase in intranasal pressure in guinea pigs. Cimetidine had no effect on this system. In passively sensitized guinea pigs (the challenge was performed 48 hr after sensitization), a 0.1-1.0 mg/kg injection of flutropium (i.v.) dose-dependently inhibited the allergic nasal mucosa capillary permeability. Atropine (10 mg/kg, i.v.) had no inhibitory action on this system. These results suggest that inhalation into the nasal cavities and i.v. injection of flutropium are effective in experimental models of drug- and allergy-induced rhinitis of the guinea pig.

Substrate specificity of 2-deoxy-scyllo-inosose synthase, the starter enzyme for 2-deoxystreptamine biosynthesis, toward deoxyglucose-6-phosphates and proposed mechanism.

A crucial enzyme in the biosynthesis of the 2-deoxystreptamine aglycon of clinically important aminocyclitol antibiotics is 2-deoxy-scyllo-inosose synthase (DOIS), which is responsible for the initial carbocycle formation of 2-deoxy-scyllo-inosose (1) from D-glucose-6-phosphate (G-6-P) (2). To get more insight into the mechanism and substrate specificity, deoxy-D-glucose-6-phosphates (deoxy-G-6-P) were chemically synthesized and subjected to the reaction with DOIS. The enzyme appeared to use 2-deoxy- and 3-deoxy-G-6-P as substrates, both of which were converted into the corresponding dideoxy-scyllo-inosose products, but 4-deoxy-G-6-P failed in cyclization by DOIS. These results clearly support the proposed reaction mechanism involving the initial oxidation at C-4 of the G-6-P substrate. Another implication is the potential use of DOIS for the preparation of useful dideoxyinososes.