First principles calculation of interface interactions and photoelectric properties of ZnSe/SnSe heterostructure.

Heterostructure engineering is an effective technology to improve photo-electronic properties of two dimensional layered semiconductors. In this paper, based on first principles method, we studied the structure, stability, energy band, and optical properties of ZnSe/SnSe heterostructure change with film layer. Results show that all heterostructures are the type-II band arrangement, and the interlayer interaction is characterized by van der Waals. The electron concentration and charge density difference implies the electron (holes) transition from SnSe to monolayer ZnSe. By increasing the layer of SnSe films, the quantum effects are weakened leading to the band gap reduced, and eventually show metal properties. The optical properties also have obvious change, the excellent absorption ability of ZnSe/SnSe heterostructures mainly near the infrared spectroscopy. These works suggest that ZnSe/SnSe heterostructure has significant potential for future optoelectronic applications.

Crohn's disease treatment and memory T-cell subset changes: insights from a case series.

Background: Crohn's disease (CD) is a chronic inflammatory bowel disease with significant morbidity, affecting millions worldwide. The intricacies of immune responses in CD, especially post-treatment, remain a vital area of exploration. While memory T (Tm)-cell subsets play a pivotal role in adaptive immunity, their specific function in patients with CD after treatment is not well-understood. This study aims to investigate the effect and function of Tm-cell subsets in these patients, addressing a crucial knowledge gap in the context of CD therapeutics. Methods: A total of eight patients diagnosed with CD were selected based on predefined inclusion criteria. All patients were treated with either anti-inflammatory agents, immunosuppressive drugs, or a combination of both. For comparison, healthy donors were enrolled based on exclusion of autoimmune or inflammatory diseases. Peripheral blood mononuclear cells (PBMCs) and lymphocytes were isolated from blood and lymph node tissue respectively. The phenotype and cytokine production of T lymphocytes from both CD patients and healthy donors were analyzed using flow cytometry. Statistical comparisons of the outcomes between CD patients and healthy donors were made using Mann-Whitney test (two-tailed) and Student t-test. Results: Post-treatment CD patients exhibited an altered T cell distribution with a notable increase in CD8+ T cells in PBMCs (P=0.0005), and altered frequencies of CD4+ and CD8+ T cells in mesenteric lymph nodes (MLNs). Tm cells showed decreased interferon-γ (IFN-γ) and tumor necrosis factor-α (TNF-α) production, with significant alterations in the frequency of IFN-γ-producing CD8+ stem cell-like Tm (Tscm) cells in lesions of the MLNs from patients with CD (CD-M-Lys) compared to healthy MLNs from patients with CD (N-M-Lys) (P=0.0152). Differences in tissue-resident Tm (Trm)-cell subset frequencies were observed between the MLNs and small intestinal mucosa in CD patients. Conclusions: The treatments with anti-inflammatory agents and/or immunosuppressive drugs have a significant effect on the frequency and function of Tm-cell subsets. Clinically, these findings suggest a potential therapeutic avenue in modulating Tm-cell responses, which might be particularly beneficial for conditions where immune response modulation is crucial. Further clinical studies are warranted to explore the full therapeutic implications of these findings.

Utility of native T1 mapping and myocardial extracellular volume fraction in patients with nonischemic dilated cardiomyopathy: A systematic review and meta-analysis.

Background: Cardiac magnetic resonance imaging (CMR) based T1 mapping and extracellular volume fraction (ECV) are powerful tools for identifying myocardial fibrosis. This systematic review and meta-analysis aims to characterize the utility of native T1 mapping and ECV in patients with non-ischemic cardiomyopathy (NICM) and to clarify the prognostic significance of elevated values. Methods: A literature search was conducted for studies reporting on use of CMR-based native T1 mapping and ECV measurement in NICM patients and their association with major adverse cardiac events (MACE), ventricular arrhythmias (VAs), and left ventricular reverse remodeling (LVRR). Databases searched included: Ovid MEDLINE, EMBASE, Web of Science, and Google Scholar. The search was not restricted to time or publication status. Results: Native T1 and ECV were significantly higher in NICM patients compared to controls (MD 78.80, 95 % CI 50.00, 107.59; p < 0.01; MD 5.86, 95 % CI 4.55, 7.16; p < 0.01). NICM patients who experienced MACE had higher native T1 and ECV (MD 52.87, 95 % CI 26.59, 79.15; p < 0.01; MD 6.03, 95 % CI 3.79, 8.26; p < 0.01). There was a non-statistically significant trend toward higher native T1 time in NICM patients who experienced VAs. NICM patients who were poor treatment responders had higher baseline native T1 and ECV (MD 40.58, 95 % CI 12.90, 68.25; p < 0.01; MD 3.29, 95 % CI 2.25, 4.33; p < 0.01). Conclusions: CMR-based native T1 and ECV quantification may be useful tools for risk stratification of patients with NICM. They may provide additional diagnostic utility in combination with LGE, which poorly characterizes fibrosis in patients with diffuse myocardial involvement.

The electronic and optical properties of Cs2BX6 (B = Zr, Hf) perovskites with first-principle method.

The electronic structures and absorption properties of Cs2BX6 halide compounds are investigated with first principle calculation and exchange correlation functional of GGA-PBE. Pressure and halogen ion doping are employed to regulate band gap. All materials suffer transition from indirect to direct band gap semiconductors but with different phase transition pressure. Structural and band structure calculating results show that the value of phase transition pressure is mainly determined by the volume of octahedron. When the volume of vacancy octahedron is much less than B-ion octahedron, the lowest band point of B-d orbitals transforms to Γ point, then the indirect semiconductors transform into direct band gap semiconductors. Calculating results of optical absorption implied that the systems have obvious blue shift, which result in the optical properties reduced. Based on suitable band gap and higher absorption coefficient, Cs2ZrI4Br2 can be an ideal candidate for perovskites solar cells.

Immune Effect of T Lymphocytes Infiltrated by Tumors on Non-Small-Cell Lung Cancer.

Lung cancer is increasing every year and it has high morbidity and mortality. Antitumor immunotherapy is a new method for the treatment of lung cancer. Currently, tumor immunotherapy mainly includes classical immunotherapy and immune-targeted therapy To explore the influence of tumor T-lymphocyte (T-cell) infiltration in non-small-cell lung cancer (NSCLC) patients, 100 NSCLC patients diagnosed and treated in Changde Second People's hospital were recruited. Patients were followed up for 3 years. The subjects were divided into a survival group (group S) and a death group (group D). The patient's pathological tissue sections were made, and the degree of T-cell infiltration was counted by H&E (Hematoxylin and eosin) staining. The infiltration degree was graded, and the positive rate of T-cell subsets was calculated by immunohistochemical staining. The 3-year positive rate was 48%, with 48 cases in group S and 52 cases in group D. The positive rate of H&E staining of group S was 100%, including 0 cases of grade 0, 5 cases of grade 1 (10.42%), 16 cases of grade 2 (33.33%), and 27 cases of grade 3 (56.25%). The positive rate of group D was 86.54%, including 4 cases of grade 0 (8.89%), 10 cases of grade 1 (22.22%), 25 cases of grade 2 (55.56%), and 6 cases of grade 3 (13.33%). The total number of T-cell infiltrates in group S was much higher than that in group D (P < 0.05). Immunohistochemical results showed that the mean positive rate of CD8+ T-cell infiltration was 72.1% in group S and 47.6% in group D, with a considerable difference (P < 0.05). No remarkable difference was found in CD4+ and CD25+ (P < 0.05). CD8+ + CD4+, CD8+/CD4+, CD25+/CD8+, CD25+/CD4+, and CD25+/(CD8+ + CD4+) positive rates were calculated, and the difference between group S and group D was substantial in CD8+ + CD4+ (P < 0.05). The results showed that T cells infiltrated by tumors had an immunosuppressive effect on tumor cells.

Microbial community composition, co-occurrence network pattern and nitrogen transformation genera response to biochar addition in cattle manure-maize straw composting.

A better understanding of the microbial group influencing nitrogen (N) dynamics and cycling in composting matrix is critical in achieving good management to alleviate N loss and improve final compost quality. This study investigated the bacterial composition, structure, co-occurrence network patterns and topological roles of N transformation in cattle manure-maize straw composting using high-throughput sequencing. The two treatments used in this experiment were cattle manure and maize straw mixture (CM) and CM with 10% biochar addition (CMB). In both treatments, the bacterial community composition varied during composting and the major phyla included Actinobacteria, Firmicutes, Proteobacteria, Bacteroidetes and Chloroflexi. The phyla Actinobacteria and Proteobacteria were more abundant in CMB treatment while Firmicutes was abundant in CM piles. The metabolic functional profiles of bacteria was predicted using the "phylogenetic investigation of communities by reconstruction of unobserved states" (PICRUSt) which revealed that except for cellular processes pathway, CMB had slight higher abundance in metabolism, genetic information processing and environmental information processing than the CM. Pearson correlation revealed more significant relationship between the important bacteria communities and N transformation in CMB piles compared with CM. Furthermore, network pattern analysis revealed that the bacterial networks in biochar amended piles are more complex and harbored more positive links than that of no biochar piles. Corresponding agreement of multivariate analyses (correlation heatmap, stepwise regression, Path and network analyses) revealed that Psychrobacter, Thermopolyspora and Thermobifida in CM while Corynebacterium_1, Thermomonospora and Streptomyces in CMB were key bacterial genera affecting NH4+-N, NO3--N and total nitrogen (TN) transformation respectively during composting process. These results provide insight into nitrogen transformation and co-occurrence patterns mediating microbes and bacterial metabolism which could be useful in enhancing compost quality and mitigating N loss during composting.

Insight to key diazotrophic community during composting of dairy manure with biochar and its role in nitrogen transformation.

Analyzing diazotrophic community may help to understand nitrogen transformation in composting and improves the final compost quality. In this study, diazotrophic community dynamics were investigated in terms of nifH gene during dairy manure and corn straw composting with biochar addition using high-throughput sequencing. Biochar decreased the diversity of diazotrophic community and altered diazotroph community structure during composting. At phylum level, Proteobacteria, Actinobacteria and Firmicutes were dominant diazotrophic communities throughout composting process. Biochar addition registered higher correlation coefficient (R) between physicochemical factors (temperature, ammonium (NH4+-N) and nitrate (NO3--N)) and diazotroph community composition. Rhodopseudomonas and Pseudoxanthomonas was the key diazotrophic communities influencing NH4+-N transformation in control (CK) and biochar compost (BC), respectively, while for NO3--N transformation Clostridium and Bradyrhizobium in CK, Azospira and Methylocystis in BC served as predominant factors. These results indicated that addition of biochar altered the key diazotroph communities influencing nitrogen transformation. Furthermore, some diazotrophs (e.g. Rhodopseudomonas, Bradyrhizobium and Azospira) affecting NH4+-N and NO3--N transformation were also observed to be mediating total nitrogen (TN). Interestingly, interactions between diazotrophic communities were observed and these interactions could also influence nitrogen transformation.

Fungal community succession under influence of biochar in cow manure composting.

This study examined the influence of biochar addition on fungal community during composting of cow manure using high-throughput sequencing. Two treatments were set up, including compost of cow manure plus 10% biochar (BC) and cow manure compost without biochar (CK). Fungal community composition varied obviously during composting in both treatments, and main fungi included Aspergillus, Myriococcum, Thermomyces, Mycothermus, Scedosporium, Cladosporium, and unclassified Microascaceae. Fungal community composition was altered by biochar during composting, especially during the thermophilic and the cooling phase, promoting Aspergillus and Myriococcum while inhibiting unclassified Microascaceae and Thermomyces. Based on linear discriminant analysis effect size analysis, common indicator groups were detected in both composts; however, specific indicator groups were also found in BC treatment, including Clavicipitaceae, Tremellales, Gibberella, and Coprinopsis. Canonical correspondence analysis (CCA) indicated that moisture content, organic matter, C/N, and pH had significant correlation (p < 0.05) with fungal composition in both treatments. However, in compost added with biochar, temperature was not an important factor affecting fungal community (p > 0.05).

Differences in distribution of functional microorganism at DNA and cDNA levels in cow manure composting.

Denitrification and nitrification processes are the two prominent pathways of nitrogen (N) transformation in composting matrix. This study explored the dynamics of denitrifying and nitrifying bacteria at different composting stages of cow manure and corn straw using functional gene sequencing at DNA and cDNA levels. Corresponding agreement among OTUs, NMDS, mental test and network analyses revealed that functional bacteria community compositions and responses to physicochemical factors were different at DNA and cDNA levels. Specifically, some OTUs were detected at the DNA level but were not observed at cDNA level, differences were also found in the distribution patterns of nitrifying and denitrifying bacteria communities at both levels. Furthermore, co-occurrence network analysis indicated that Pseudomonas, Paracoccus and Nitrosomonas were identified as the keystone OTUs at the DNA level, while Paracoccus, Agrobacterium and Nitrosospira were keystone OTUs at the cDNA level. Mantel test revealed that TN, C/N and moisture content significantly influenced both the denitrifying bacteria and ammonia-oxidizing bacteria (AOB) communities at the DNA level. NO3--N, NH4+-N, TN, C/N, and moisture content only registered significant correlation with the nosZ-type denitrifiers and ammonia-oxidizing bacteria (AOB) communities at the cDNA level. Structural equation model (SEM) showed that TN, NH4+-N, and pH were direct and significantly influenced the gene abundance of denitrifying bacteria. Howbeit, TN, NH4+-N, and NO3--N had significant direct effects on amoA gene abundance.

Biochar reduces nutrient loss and improves microbial biomass of composted cattle manure and maize straw.

In this study, nutrient loss, the direct and indirect relationship between period, compost types, temperature, total nitrogen (TN), nitrate nitrogen (NO3 - -N), ammonium nitrogen (NH4 + -N), microbial biomass carbon (MBC), and microbial biomass nitrogen (MBN) were investigated during composting of cattle manure-maize straw mixture. This study findings revealed that biochar addition lowered NH4 + -N but did not increase NO3 - -N concentrations unlike no biochar piles during composting. The first-order kinetic models showed that biochar accelerated organic matter (OM) degradation and improved nitrogen mineralization, consequently reducing TN losses by 13.6% and OM losses by 12.66%. Transformation ratio of MBC/MBN, coupled with other chemical components of the entire microbial community, suggested a shift in the microbial succession and diversity during composting from the dominant bacteria and actinomycetes to fungi. The structural equation model and path coefficient revealed temperature to be the main factor mediating the evolution of MBC and MBN in composting. The physicochemical variables, phytotoxicity, and final product quality revealed that biochar incorporation to the composting feedstock is an ideal material for mitigating problems of TN and OM losses in composting and ultimately enhancing the fertility potential of the final compost product.

Genetic associations as indices for assessing nitrogen transformation processes in co-composting of cattle manure and rice straw.

In this study, mechanism of nitrogen transformation was investigated in terms of genetic associations (nitrogen-related gene groups) in co-composting of cattle manure and rice straw. Mutual validation among KEGG, Pearson correlation, stepwise regression, and Path analyses indicated that the functional genes synergistically affected on nitrogen transformation in composting process. NxrA/qnorB (0.9419 ±â€¯0.0334) and (amoA + anammox)/Bacteria (0.7187 ±â€¯0.0334) were the key functional gene groups mediating NH4+-N transformation. AmoA/(narG + napA) (-0.8400 ±â€¯0.0129), amoA/bacteria (0.8692 ±â€¯0.0273), and (nirK + nirS)/nosZ (1.1652 ±â€¯0.0089) determined NO3--N, NO2--N and N2O transformation, respectively. AmoA/(napA + narG) mediated both NO3--N and NO2--N transformation. AmoA/anammox (-0.7172 ±â€¯0.0591) and (nirK + nirS)/nosZ (-0.6626 ±â€¯0.0825) served as predominant factors for total nitrogen removal. These results provided a molecular-level insight that nitrification, anaerobic ammonia oxidation and denitrification (SNAD) might simultaneously contribute to nitrogen transformation during composting, rather than sequentially.

Microbial Community Succession and Response to Environmental Variables During Cow Manure and Corn Straw Composting.

In composting system, the composition of microbial communities is determined by the constant change in the physicochemical parameters. This study explored the dynamics of bacterial and fungal communities during cow manure and corn straw composting using high throughput sequencing technology. The relationships between physicochemical parameters and microbial community composition and abundance were also evaluated. The sequencing results revealed the major phyla included Proteobacteria, Bacteroidetes, Firmicutes, Chloroflexi and Actinobacteria, Ascomycota, and Basidiomycota. Linear discriminant analysis effect size (LEfSe) illustrated that Actinomycetales and Sordariomycetes were the indicators of bacteria and fungi in the maturation phase, respectively. Mantel test showed that NO3 --N, NH4 +-N, TN, C/N, temperature and moisture content significantly influenced bacterial community composition while only TN and moisture content had a significant effect on fungal community structure. Structural equation model (SEM) indicated that TN, NH4 +-N, NO3 --N and pH had a significant effect on fungal abundance while TN and temperature significantly affected bacterial abundance. Our finding increases the understanding of microbial community succession in cow manure and corn straw composting under natural conditions.

Bacterial community succession in dairy manure composting with a static composting technique.

This study applied high-throughput sequencing technology and PICRUSt (phylogenetic investigation of communities by reconstruction of unobserved state) to examine the microbial population dynamics during the composting of dairy manure and rice straw in a static (without turning) composting system. The results showed that the composition of the bacterial community varied significantly during the composting process. The dominant phyla included Firmicutes, Proteobacteria, Bacteroidetes, Actinobacteria, and Chloroflexi. Biodiversity indices showed that bacterial community diversity had the peak value during the mesophilic phase. Redundancy analysis indicated that nitrogen was the most important factor in the distribution of genera during the composting process. Finally, the Pearson correlation results suggested that Thermomonospora and Thermopolyspora could be the biomarkers of the composting maturation phase. The metabolic characteristics of the bacterial communities were predicted by PICRUSt. The result showed that metabolism of amino acids, lipids, and most of the carbohydrates increased during the whole composting treatment. However, methane metabolism, carbon fixation pathways in prokaryotes, and nucleotide metabolism decreased after the thermophilic phase. The present study provides a better understanding for bacterial community composition and function succession in dairy manure composting.

In-Hospital Outcomes After Transcatheter Versus Surgical Aortic Valve Replacement in Octogenarians.

Background Octogenarians have low physiologic reserve and may benefit more from transcatheter aortic valve replacement ( TAVR ) than surgical aortic valve replacement ( SAVR ). Methods and Results This retrospective cohort study based on the National Inpatient Sample included octogenarians who underwent TAVR or SAVR from 2012 to 2015. Crude and standardized-morbidity-ratio-weighted regression models were used to compare in-hospital outcomes. Among 19 145 TAVR and 9815 SAVR hospitalizations, TAVR patients had higher Charlson Comorbidity Index ( CCI ) scores (2.0 versus 0.8, P<0.0001) than SAVR patients. Before weighting, TAVR was associated with significantly shorter length of stay, more home discharges, and lower incidences of acute kidney injury, bleeding, and cardiogenic shock. Associations were consistent across Charlson Comorbidity Index, except for TAVR being associated with greater length of stay reductions among patients with Charlson Comorbidity Index ≥2, compared with Charlson Comorbidity Index <2 (change in estimate -3.56 versus -2.61 days, P=0.004). After weighting, TAVR patients had significantly shorter length of stay (change in estimate -3.29 days, 95% CI -3.82, -2.75) and lower odds of transfer to skilled nursing facility (odds ratio 0.34, 95% CI 0.29, 0.41), acute kidney injury (odds ratio 0.55, 95% CI 0.45, 0.68), bleeding (odds ratio 0.44, 95% CI 0.37, 0.53), and cardiogenic shock (odds ratio 0.55, 95% CI 0.33, 0.92), compared with SAVR patients. Odds of permanent pacemaker implantation, transient ischemic attack/stroke, vascular complications, and in-hospital mortality were not significantly different. Conclusions TAVR may be preferred over SAVR in high-risk octogenarians because of shorter length of stay, better discharge disposition, and less acute kidney injury, and bleeding. All octogenarians may benefit more from TAVR , irrespective of comorbidity burden, but additional research is needed to confirm our findings.

Relation of an Echocardiographic-Based Cardiac Calcium Score to Mitral Stenosis Severity and Coronary Artery Disease in Patients with Severe Aortic Stenosis.

Patients with calcific aortic stenosis (AS) often have diffuse cardiac calcification involving the mitral valve apparatus and coronary arteries. We examined the association between global cardiac calcification quantified by a previously validated echocardiographic calcium score (eCS) with the severity of mitral stenosis (MS) and coronary artery disease (CAD) in patients with a clinical diagnosis of severe calcific AS. In this sample of 147 patients (mean age 81 ± 9 years, 50% male), 81 patients (55%) were determined by echocardiography to have some degree of MS. Higher mean eCS was observed in patients with more severe MS (r = 0.54, p < 0.0001). Higher eCS was also inversely associated with mitral valve area (r = -0.31, p = 0.001) and positively associated with mitral valve mean pressure gradient (r = 0.46, p < 0.0001) and mitral valve peak flow velocity (r = 0.55, p < 0.0001). The area under the receiver operating characteristic curve for using eCS to predict the presence of MS was 0.76. An eCS ≥ 8 predicted MS with a sensitivity of 68%, specificity of 76%, positive predictive value of 77%, and negative predictive value of 66%. High eCS, relative to low eCS, was associated with 2.70 times the adjusted odds of CAD (odds ratio = 2.70, 95% confidence interval 1.02 to 7.17). In conclusion, global cardiac calcification is associated with MS and CAD in patients with severe calcific AS, and eCS shows ability to predict the presence of MS. This study suggests that a simple eCS may be used as part of a risk-stratification tool in patients with severe calcific aortic valve stenosis.

CTDP-32476: A Promising Agonist Therapy for Treatment of Cocaine Addiction.

Agonist-replacement therapies have been successfully used for treatment of opiate and nicotine addiction, but not for cocaine addiction. One of the major obstacles is the cocaine-like addictive potential of the agonists themselves. We report here an atypical dopamine (DA) transporter (DAT) inhibitor, CTDP-32476, that may have translational potential for treating cocaine addiction. In vitro ligand-binding assays suggest that CTDP-32476 is a potent and selective DAT inhibitor and a competitive inhibitor of cocaine binding to the DAT. Systemic administration of CTDP-32476 alone produced a slow-onset, long-lasting increase in extracellular nucleus accumbens DA, locomotion, and brain-stimulation reward. Drug-naive rats did not self-administer CTDP-32476. In a substitution test, cocaine self-administration rats displayed a progressive reduction in CTDP-32476 self-administration with an extinction pattern of drug-taking behavior, suggesting significantly lower addictive potential than cocaine. Pretreatment with CTDP-32476 inhibited cocaine self-administration, cocaine-associated cue-induced relapse to drug seeking, and cocaine-enhanced extracellular DA in the nucleus accumbens. These findings suggest that CTDP-32476 is a unique DAT inhibitor that not only could satisfy 'drug hunger' through its slow-onset long-lasting DAT inhibitor action, but also render subsequent administration of cocaine ineffectual-thus constituting a novel and unique compound with translational potential as an agonist therapy for treatment of cocaine addiction.

Xenon neuroprotection in experimental stroke: interactions with hypothermia and intracerebral hemorrhage.

BACKGROUND: Xenon has been proven to be neuroprotective in experimental brain injury. The authors hypothesized that xenon would improve outcome from focal cerebral ischemia with a delayed treatment onset and prolonged recovery interval. METHODS: Rats were subjected to 70 min temporary focal ischemia. Ninety minutes later, rats were treated with 0, 15, 30, or 45% Xe for 20 h or 0 or 30% Xe for 8, 20, or 44 h. Outcome was measured after 7 days. In another experiment, after ischemia, rats were maintained at 37.5° or 36.0°C for 20 h with or without 30% Xe. Outcome was assessed 28 days later. Finally, mice were subjected to intracerebral hemorrhage with or without 30% Xe for 20 h. Brain water content, hematoma volume, rotarod function, and microglial activation were measured. RESULTS: Cerebral infarct sizes (mean±SD) for 0, 15, 30, and 45% Xe were 212±27, 176±55, 160±32, and 198±54 mm, respectively (P=0.023). Neurologic scores (median±interquartile range) followed a similar pattern (P=0.002). Infarct size did not vary with treatment duration, but neurologic score improved (P=0.002) at all xenon exposure durations (8, 20, and 44 h). Postischemic treatment with either 30% Xe or subtherapeutic hypothermia (36°C) had no effect on 28-day outcome. Combination of these interventions provided long-term benefit. Xenon improved intracerebral hemorrhage outcome measures. CONCLUSION: Xenon improved focal ischemic outcome at 7, but not 28 days postischemia. Xenon combined with subtherapeutic hypothermia produced sustained recovery benefit. Xenon improved intracerebral hemorrhage outcome. Xenon may have potential for clinical stroke therapy under carefully defined conditions.