Dissociable roles of thalamic nuclei in the refinement of reaches to spatial targets.

Reaches are complex movements that are critical for survival, and encompass the control of different aspects such as direction, speed, and endpoint precision. Complex movements have been postulated to be learned and controlled through distributed motor networks, of which the thalamus is a highly connected node. Still, the role of different thalamic circuits in learning and controlling specific aspects of reaches has not been investigated. We report dissociable roles of two distinct thalamic nuclei - the parafascicular (Pf) and ventroanterior/ventrolateral (VAL) nuclei - in the refinement of spatial target reaches in mice. Using 2-photon calcium imaging in a head-fixed joystick task where mice learned to reach to a target in space, we found that glutamatergic neurons in both areas were most active during reaches early in learning. Reach-related activity in both areas decreased late in learning, as movement direction was refined and reaches increased in accuracy. Furthermore, the population dynamics of Pf, but not VAL, covaried in different subspaces in early and late learning, but eventually stabilized in late learning. The neural activity in Pf, but not VAL, encoded the direction of reaches in early but not late learning. Accordingly, bilateral lesions of Pf before, but not after learning, strongly and specifically impaired the refinement of reach direction. VAL lesions did not impact direction refinement, but instead resulted in increased speed and target overshoot. Our findings provide new evidence that the thalamus is a critical motor node in the learning and control of reaching movements, with specific subnuclei controlling distinct aspects of the reach early in learning.

Exploration biases how forelimb reaches to a spatial target are learned.

The brain can learn to generate actions, such as reaching to a target, using different movement strategies. Understanding how different variables bias which strategies are learned to produce such a reach is important for our understanding of the neural bases of movement. Here we introduce a novel spatial forelimb target task in which perched head-fixed mice learn to reach to a circular target area from a set start position using a joystick. These reaches can be achieved by learning to move into a specific direction or to a specific endpoint location. We find that mice gradually learn to successfully reach the covert target. With time, they refine their initially exploratory complex joystick trajectories into controlled targeted reaches. The execution of these controlled reaches depends on the sensorimotor cortex. Using a probe test with shifting start positions, we show that individual mice learned to use strategies biased to either direction or endpoint-based movements. The degree of endpoint learning bias was correlated with the spatial directional variability with which the workspace was explored early in training. Furthermore, we demonstrate that reinforcement learning model agents exhibit a similar correlation between directional variability during training and learned strategy. These results provide evidence that individual exploratory behavior during training biases the control strategies that mice use to perform forelimb covert target reaches.

Evaluation of at-home methods for N95 filtering facepiece respirator decontamination.

N95 filtering facepiece respirators (FFRs) are essential for the protection of healthcare professionals and other high-risk groups against Coronavirus Disease of 2019 (COVID-19). In response to shortages in FFRs during the ongoing COVID-19 pandemic, the Food and Drug Administration issued an Emergency Use Authorization permitting FFR decontamination and reuse. However, although industrial decontamination services are available at some large institutions, FFR decontamination is not widely accessible. To be effective, FFR decontamination must (1) inactivate the virus; (2) preserve FFR integrity, specifically fit and filtering capability; and (3) be non-toxic and safe. Here we identify and test at-home heat-based methods for FFR decontamination that meet these requirements using common household appliances. Our results identify potential protocols for simple and accessible FFR decontamination, while also highlighting unsuitable methods that may jeopardize FFR integrity.

[End tidal PCO(2) for evaluation of severity of disease in idiopathic pulmonary arterial hypertension and chronic thromboembolic pulmonary hypertension].

Objective: To explore the difference in end tidal PCO(2) (P(ET)CO(2)) between idiopathic pulmonary arterial hypertension (IPAH) and chronic thromboembolic pulmonary hypertension (CTEPH), and to analyze the correlation between P(ET)CO(2) and the indexes of disease severity in IPAH and CTEPH patients. Methods: Data were retrieved from 68 IPAH patients and 52 CTEPH patients who all had received right-heart catheterization, pulmonary function test and cardiopulmonary exercise testing at Shanghai Pulmonary Hospital from October 2011 to October 2014. In addition, other clinical parameters were also collected. Results: The IPAH group had a significantly higher mPAP, PVR [(60±16) mmHg (1 mmHg=0.133 kPa), (13±6) Wood U ] than the CTEPH group [(46±12) mmHg, (9±4) Wood U, t=4.90, 4.83, all P<0.01]. Meanwhile, the IPAH group had a lower percentage of predicted peakVO(2), oxygen pulse [(45±15)%, (60±22)%] compared with the CTEPH group [(53±16)%, (68±21)%, t=-2.42, -1.96, all P<0.05]. The value of P(ET)CO(2) at rest, AT, peak in the IPAH patients [(27±5), (28±7), (25±7) mmHg] were higher than those in the CTEPH patients [(24±4) mmHg, (23±6) mmHg, (21±6) mmHg, t=3.22-4.54, all P<0.01]. There was a significantly difference in P(ET)CO(2) at AT and peak between WHO-FC Ⅰ-Ⅱ and Ⅲ-Ⅳ subgroups in IPAH (t=2.55, 2.60, all P<0.05) and CTEPH (t=2.39, P<0.05), except for P(ET)CO(2) at peak in the CTEPH patients (t=1.71, P>0.05). A moderately inverse correlation was found between P(ET)CO(2) at AT and NT-proBNP in the IPAH group (r=-0.58, P<0.01), meanwhile P(ET)CO(2) at AT in the CTEPH group was weakly correlated with NT-proBNP (r=-0.34, P<0.05). Conclusions: Compared with the CTEPH patients, the IPAH patients had significantly decreased exercise capacity and increased P(ET)CO(2). P(ET)CO(2) could reflect the disease severity in both IPAH and CTEPH patients, being superior in IPAH than in CTEPH. Furthermore, P(ET)CO(2) at AT might be better than P(ET)CO(2) at peak in reflecting the ventilatory efficiency.

As2O3 induces apoptosis of the human B lymphoma cell line MBC-1.

OBJECTIVE: To see how arsenic trioxide (As2O3) affects proliferation of the human B lymphoma cell line MBC-1. METHODS: We studied the effect of As2O3 on MBC-1 cells and its mechanism by morphological observation, flow cytometry assay and DNA gel electrophoresis. RESULTS: As2O3 could upregulate p53 gene expression at protein level, inducing cell apoptosis and inhibiting the proliferation of MBC-1 cells. Upregulation of p53 expression appears to be important in the apoptosis of MBC-1 cells. CONCLUSIONS: As2O3 can inhibit the proliferation of MBC-1 cells by upregulating p53 gene expression, thus inducing apoptosis.

LOH and mutational analysis of p53 alleles in mouse urinary bladder carcinomas induced by N-butyl-N-(4-hydroxybutyl) nitrosamine.

In human urinary bladder carcinogenesis, alterations in the p53 tumor suppressor gene are common events. We have previously reported that they are also frequent in invasive urinary bladder carcinomas induced by N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) in NON/Shi mice. To further investigate the significance of the p53 gene status for mouse urinary bladder carcinogenesis, we examined both allele loss and mutational alterations in urinary bladder cancers of (NON/Shi x C3H/He/Shi) F1 hybrid mice exposed to the carcinogen for 12 weeks and then maintained for a further 9 weeks without treatment. An intragenic silent polymorphism within exon 7 of the p53 gene between NON/Shi and C3H/He/Shi mice allows assessment of allele loss of the p53 gene and determination of the parental origin of mutated and/or lost alleles. A tissue microdissection method was employed to obtain carcinoma samples without excessive contamination with normal tissue. Allele losses were detected in one of 14 tumors (7.1%) and nine mutations in eight of 14 (57%) tumors were found in exons 5-8 by polymerase chain reaction-single strand conformation polymorphism followed by DNA direct sequencing analysis. All mutations involved one base substitution with an amino acid change, although the types of base substitution were random. In conclusion, the high incidence of p53 alterations suggests a significant role in the genesis of invasive urinary bladder tumors in BBN-treated mice.

Promotion by sodium L-ascorbate in rat two-stage urinary bladder carcinogenesis is dependent on the interval of administration.

In our two-stage model of rat urinary bladder carcinogenesis employing N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN) as the initiator, sodium L-ascorbate (Na-AsA) exhibits dose-dependent promotion. In the present study, in order to assess the possible reversibility of the promoting effects, we investigated how different administration periods of Na-AsA influence its promoting activity. In experiment 1, rats were treated with 5% Na-AsA for different administration periods with or without withdrawal and injected with 5-bromo-2'-deoxyuridine (BrdU) to allow determination of the cell proliferation status. Replicative DNA synthesis in the urinary bladder epithelium was shown to return to normal after removal of the promoting stimulus. In experiment 2, rats were initially given BBN for 4 weeks and subsequently received 16 weeks of Na-AsA, alternating with basal diet, at intervals of 4, 8 or 16 weeks, within a total 32-week period. The longer the continuous exposure to Na-AsA, the greater the yield of papillomas and carcinomas in the urinary bladder. In experiment 3, Na-AsA was given for 4 or 8 weeks after BBN initiation and the animals were killed at weeks 8 and 12. Both promotion of lesion development and increase of DNA synthesis in the urinary bladder epithelium were dependent on the length of exposure to Na-AsA and the total period of exposure. The results indicate that the promoting effects of Na-AsA in urinary bladder carcinogenesis are reversible to a certain extent after its withdrawal, and the existence of a cumulative exposure time threshold seems likely.

Expression of HSP70 gene in rat brain after exposures to repeated +Gz.

OBJECTIVE: To study the changes of mRNA expression of heat shock protein 70 (HSP70) in the rat brain exposed to repeated +Gz. METHOD: The mRNA expression levels of HSP70 in rat brain were measured by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). RESULT: The HSP70 mRNA expression levels in rat brains taken 30 min and 6 h after repeated +Gz exposures were significantly higher than those in control group, while the difference between the levels of control group and those of experimental rat brains taken 24 h after +Gz exposure was not significant. CONCLUSION: It is suggested that HSP70 mRNA expression in rat brain can be induced by repeated +Gz exposures and the increased HSP70 mRNA expression may play an important role in self-protection against brain damage induced by +Gz exposures.

Concentration dependent promoting effects of sodium L-ascorbate with the same total dose in a rat two-stage urinary bladder carcinogenesis.

Sodium L-ascorbate (Na-AsA) has been demonstrated to be a strong promoter of rat urinary bladder tumor development initiated by N-butyl-N-(4-hydroxybutyl)nitrosamine (BBN). In the present study, we investigated variation in its promoting activity when the same total dose was given with different concentrations and exposure times. After 4 weeks administration of 0.05% BBN, group 1 served as a control without any post-initiation treatment. The rats in groups 2-4 received 1.25% Na-AsA diet for 36 weeks, 2.5% Na-AsA for 18 weeks and 5% Na-AsA for 8 weeks, respectively. Tumor number (papillomas and carcinomas) was greatest in group 3, and area in group 4 (P < 0.05). However, no enhancement was noted in group 2, although preneoplastic lesions were significantly increased. These results suggest that with the same total administration dose, high concentration of Na-AsA has the strongest promoting effects on tumor development in urinary bladder carcinogenesis.

Inhibition of interleukin-2 receptor (CD25) expression induced on T cells from children with acute lymphoblastic leukemia.

Peripheral blood lymphocytes obtained from children with acute lymphoblastic leukemia (ALL) at onset were studied for the expression of interleukin-2 (IL-2) receptor alpha-chain (CD25) by two-color flow-cytometric analysis. Stimulated with anti-CD3 monoclonal antibody (mAb) alone. CD25 expression was significantly suppressed in CD4+ T cells from 27 of 48 (56.3%) cases and in CD8+ T cells from 29 of 48 (60.4%) cases. When stimulated with anti-CD3 mAb plus phorbol 12-myristate 13-acetate (PMA), CD25 expression was clearly restored in certain cases of ALL. When PMA plus ionomycin were used for stimulation of T cells. CD25 was inducible in a majority of cases. Interestingly CD25 expression upon anti-CD3 mAb stimulation was recovered after complete remission had been achieved. These observations suggest the presence in ALL children at onset of an in vitro defect in the signal transduction pathway of the T-cell-receptor/CD3 complex, resulting in inefficient CD25 expression. However, immune-staining analysis indicated that protein kinase C was normally translocated from the cytosol fraction to the cell membrane fraction. The mobilization of cytoplasmic free calcium is also normal.

Fas/APO-1 (CD95)-mediated cytotoxicity is responsible for the apoptotic cell death of leukaemic cells induced by interleukin-2-activated T cells.

Apoptotic cell death is induced by the cross-linking of Fas/APO-1 receptor (CD95) in acute myelogenous leukaemia (AML) cells. Since CD95 ligand (CD95L) is expressed on interleukin-2 (IL-2)-activated T cells, we investigated the involvement of CD95-CD95L pathway in T cell-mediated cytotoxicity against AML cells. Activated CD8+ T cells could efficiently kill a parental CD95-sensitive AML cell line, MML-1 and, to a lesser extent, a CD95-resistant clone, MML-1R. Neither MML-1 nor MML-1R cells were killed by activated CD4+ T cells. The blocking monoclonal antibody (MoAb) against CD95, ZB4, caused a significant inhibition of T-cell-mediated cytotoxicity against MML-1 cells but not against MML-1R cells. Interestingly, MML-1 cells underwent the classic nuclear morphologic changes and DNA fragmentation characteristic of apoptosis when cultured with activated T cells. Enumeration of apoptotic and necrotic nuclei showed that both apoptosis and necrosis were induced in MML-1 cells, whereas necrosis was exclusively observed in MML-1R cells. Apoptosis of MML-1 cells was completely blocked in the presence of ZB4 MoAb. Similarly, blocking by ZB4 MoAb significantly inhibited T-cell-mediated lysis of fresh AML cells in a CD95-sensitive group, but not in a CD95-refractory group. In addition CD8+ T cells expressed CD95L mRNA more abundantly than CD4+ T cells upon activation by IL-2. These findings suggest that T-cell-mediated cytotoxicity against AML cells requires participation of CD95-CD95L pathway for cytotoxic signal transduction leading to target apoptosis.