Tolerance Analysis Research Articles

A metallothionein from disk abalone (Haliotis discus discus): Insights into its functional roles in immune response, metal tolerance, and oxidative stress

Metallothioneins (MTs) are cysteine-rich metal-binding proteins whose expression is induced by exposure to essential and non-essential metals, making them potential biological markers for assessing metal pollution in various biomonitoring programs. However, the functional properties of these proteins are yet to be comprehensively characterized in most marine invertebrates. In this study, we identified and characterized an MT homolog from the disk abalone (Haliotis discus discus), referred to as disk abalone MT (AbMT). AbMT exhibited the same primary structural features as MTs from other mollusks containing two β-domains (β2β1-form). AbMT protein demonstrated metal-binding and detoxification abilities against Zn, Cu, and Cd, as evidenced by Escherichia coli growth kinetics, metal tolerance analysis, and UV absorption spectrum. Transcriptional analysis revealed that AbMT was ubiquitously expressed in all analyzed tissues and upregulated in gill tissue following challenge with Vibrio parahaemolyticus, Listeria monocytogenes, and viral hemorrhagic septicemia virus (VHSV). Additionally, overexpression of AbMT suppressed LPS-induced NO production in RAW264.7 macrophages, protected cells against H2O2-induced oxidative stress, and promoted macrophage polarization toward the M1 phase. Conclusively, these findings suggest an important role for AbMT in environmental stress protection and immune regulation in disk abalone.

GLP-1 receptor agonist improves metabolic disease in a pre-clinical model of lipodystrophy.

Individuals with lipodystrophies typically suffer from metabolic disease linked to adipose tissue dysfunction including lipoatrophic diabetes. In the most severe forms of lipodystrophy, congenital generalised lipodystrophy, adipose tissue may be almost entirely absent. Better therapies for affected individuals are urgently needed. Here we performed the first detailed investigation of the effects of a glucagon like peptide-1 receptor (GLP-1R) agonist in lipoatrophic diabetes, using mice with generalised lipodystrophy. Lipodystrophic insulin resistant and glucose intolerant seipin knockout mice were treated with the GLP-1R agonist liraglutide either acutely preceding analyses of insulin and glucose tolerance or chronically prior to metabolic phenotyping and ex vivo studies. Acute liraglutide treatment significantly improved insulin, glucose and pyruvate tolerance. Once daily injection of seipin knockout mice with liraglutide for 14 days led to significant improvements in hepatomegaly associated with steatosis and reduced markers of liver fibrosis. Moreover, liraglutide enhanced insulin secretion in response to glucose challenge with concomitantly improved glucose control. GLP-1R agonist liraglutide significantly improved lipoatrophic diabetes and hepatic steatosis in mice with generalised lipodystrophy. This provides important insights regarding the benefits of GLP-1R agonists for treating lipodystrophy, informing more widespread use to improve the health of individuals with this condition.

ISOLATION AND CHARACTERIZATION OF YEAST FROM FERMENTED SEMERU ARABICA WINE COFFEE CHERRIES

Wine coffee is fermented coffee cherries with a wine-like flavor and aroma. The purpose of this study was to isolate, characterize tolerantand thermotolerant yeast from Semeru Arabica wine coffee cherries and to determine the molecular identification of the selected yeast. In this study, the Semeru Arabica coffee cherries were fermented for 30 days.Yeast isolation was performed on samples collected each week during fermentation. The selected isolates were characterized for macroscopic and microscopic morphology, followed by ethanol and temperature tolerance analysis. The isolate with the most desirable characteristics, designated CW30, grew well at 30ºC but was sensitive to temperatures above 35ºC. The isolate grew well in media containing 5% ethanol but was sensitive to a 10% ethanol concentration. Based on nucleotide sequence analysis, CW30 exhibited 100% similarity with Candida ethanolica isolate 3-1-19.

An improved tolerance analysis method for gear mechanisms considering form deviations and elastic-plastic contact behaviors

The contact quality between geometric features significantly affects the assembly accuracy and dynamic performance of mechanical products. However, accurate and efficient contact problem solving is currently a common challenge in the fields of tolerance analysis and tooth contact analysis (TCA). In this paper, a tolerance analysis method for gear mechanisms considering form deviations and elastic-plastic contact behaviors is proposed. Specifically, tolerance modeling of systematic and random deviations on tooth surfaces is represented by skin model shapes (SMS). Elastic-plastic contact behaviors including contact deformation, status, and stress distribution are modeled and calculated by a modified boundary element method (BEM). The accuracy and efficiency of this BEM algorithm are verified by comparing with the finite element method (FEM), indicating that the BEM can greatly improve the computational efficiency while maintaining high solution accuracy. A case study for a pair of spur gears demonstrates the significant effects of contact behaviors on transmission performance. This research provides an effective method to introduce mechanical contact behaviors in tolerance analysis of gear mechanisms. In addition, the contact simulation results can also be adopted in the analysis of vibration, reliability, and other aspects.

Perampanel for Treatment of People with a Range of Epilepsy Aetiologies in Clinical Practice: Evidence from the PERMIT Extension Study.

It is important to assess the effectiveness of an antiseizure medication in treating different epilepsy aetiologies to optimise individualised therapeutic approaches. Data from the PERaMpanel pooled analysIs of effecTiveness and tolerability (PERMIT) Extension study were used to assess the effectiveness and safety/tolerability of perampanel (PER) when used to treat individuals with a range of epilepsy aetiologies in clinical practice. A post hoc analysis was conducted of PERMIT Extension data from individuals with a known aetiology. Retention was assessed after 3, 6 and 12months. Effectiveness was assessed after 3, 6 and 12months and at the last visit (last observation carried forward). Effectiveness assessments included responder rate (≥ 50% seizure frequency reduction) and seizure freedom rate (no seizures since at least the prior visit). Safety/tolerability was assessed by evaluating adverse events (AEs) and AEs leading to discontinuation. PERMIT Extension included 1945 individuals with structural aetiology, 1012 with genetic aetiology, 93 with an infectious aetiology, and 26 with an immune aetiology. Retention rates at 12months were 61.1% (structural), 65.9% (genetic), 56.8% (infectious) and 56.5% (immune). At the last visit, responder rates (total seizures) were 43.3% (structural), 68.3% (genetic), 37.0% (infectious) and 20.0% (immune), and corresponding seizure freedom rates were 15.8%, 46.5%, 11.1% and 5.0%, respectively. AE incidence rates were 58.0% (structural), 46.5% (genetic), 51.1% (infectious) and 65.0% (immune), and corresponding rates of discontinuation due to AEs over 12months were 18.9%, 16.4%, 18.5% and 21.7%, respectively. The types of AEs reported were generally consistent across aetiology subgroups, with no idiosyncratic AEs emerging. Although PER was effective and generally well tolerated when used to treat individuals with a range of epilepsy aetiologies in clinical practice, variability in its effectiveness and tolerability across the subgroups indicates that PER may be particularly useful for individuals with specific epilepsy aetiologies.

Gene expression analysis of drought tolerance and cuticular wax biosynthesis in diploid and tetraploid induced wallflowers

Whole-genome doubling leads to cell reprogramming, upregulation of stress genes, and establishment of new pathways of drought stress responses in plants. This study investigated the molecular mechanisms of drought tolerance and cuticular wax characteristics in diploid and tetraploid-induced Erysimum cheiri. According to real-time PCR analysis, tetraploid induced wallflowers exhibited increased expression of several genes encoding transcription factors (TFs), including AREB1 and AREB3; the stress response genes RD29A and ERD1 under drought stress conditions. Furthermore, two cuticular wax biosynthetic pathway genes, CER1 and SHN1, were upregulated in tetraploid plants under drought conditions. Leaf morphological studies revealed that tetraploid leaves were covered with unique cuticular wax crystalloids, which produced a white fluffy appearance, while the diploid leaves were green and smooth. The greater content of epicuticular wax in tetraploid leaves than in diploid leaves can explain the decrease in cuticle permeability as well as the decrease in water loss and improvement in drought tolerance in wallflowers. GC‒MS analysis revealed that the wax components included alkanes, alcohols, aldehydes, and fatty acids. The most abundant wax compound in this plant was alkanes (50%), the most predominant of which was C29. The relative abundance of these compounds increased significantly in tetraploid plants under drought stress conditions. These findings revealed that tetraploid-induced wallflowers presented upregulation of multiple drought-related and wax biosynthesis genes; therefore, polyploidization has proved useful for improving plant drought tolerance.

Characterization of the damage mechanism in CFRP composites under mode I based on comprehensive analysis of AE signals

This study utilises several methods centred on acoustic emission (AE) waveform characteristics to conduct in-depth analysis of the damage mechanism of composite materials under mode I. Combined with the peak frequency statistics and continuous wavelet transform (CWT) of AE signals, three damage modes of laminated composites during loading are identified. The cumulative energy of three damage modes separated by variational modal decomposition (VMD) is used to characterize their damage evolution process. Based on the findings, delamination and fiber–matrix debonding are proven to be the dominant damage mechanisms for mode I. The comprehensive analysis results from AE's accumulated energy, peak frequency distribution, CWT, VMD, and sentry function can accurately detect the onset time of delamination in laminates. The equivalent crack length is calculated by the load value determined at the initial time of delamination, and the strain energy release rate calculated by the load value and crack length at this time is used as the interlaminar fracture toughness value of mode I. This research method will provide a new idea for the damage tolerance analysis of mode I delamination in laminated structures.

Optical MTF and aberration predictions with the Helmholtz-Lagrange invariant

The paper presents a method for a rough estimation of the aberrations and of the MTF of an optical system by means of the Helmholtz-Lagrange invariant. The method allows for a rough understanding of an optical design and can find application in those cases where approximated performance predictions are needed, for example, before running complex and time-consuming analyses. The simple approach presented here is a rough approximation and should always be followed by the usual tolerance analyses.

Isolation and identification of non‐Saccharomyces yeasts and their flavour characteristics while brewing Yinhong plum wine

AbstractThe objective of this study was to screen non‐Saccharomyces yeast suitable for enhancing the aroma of Yinhong plum wine. Five non‐Saccharomyces yeasts with strong ester‐producing ability, namely, Wa3 (Wickerhamomyces anomalus), Hu12 (Hanseniaspora uvarum), Pk2A2, PkW2 and PkY2 (Pichia kluyveri), were selected from various naturally fermented fruit juices for molecular biological identification, analyses of physiological characteristics and tolerance and determination of their effects on the flavour of Yinhong plum wine. Analysis of brewing characteristics showed that they differed in their abilities of producing acetic acid, hydrogen sulfide, killer activity and tolerance. Among them, Wa3 showed the best tolerance to high concentrations of alcohol, sugar and SO2. Compared to the control Saccharomyces cerevisiae FX10, Yinhong plum wine fermented with non‐Saccharomyces yeast exhibited a decrease in alcohol content, while residual sugar, glycerol content and the types and quantities of volatile compounds all showed an increase. Among the fermented wines, PkY2 generated the highest content of ester compounds in the fermented wine, which was 5.66 times higher than that generated by the control. The compounds produced by PkY2 were closely related to various ethyl and acetate odour‐active compounds (odour‐active value >1), which could impart a rich floral and fruity aroma to Yinhong plum wine. Sensory analysis revealed that both floral and fruity aromas were significantly increased in Yinhong plum wine fermented by non‐Saccharomyces yeasts, with the highest scores of floral and fruity aromas for PkY2. In conclusion, a Pichia kluyveri strain, PkY2, with excellent brewing characteristics and potential for enhancing the aroma of Yinhong plum wine was identified.

Real-world Data on Ocrelizumab Treatment: Efficacy, Safety, and Tolerability Analysis in Patients with Relapsing Remitting and Primary Progressive Multiple Sclerosis in Southeastern Ontario (P2-6.014)

Real-world Data on Ocrelizumab Treatment: Efficacy, Safety, and Tolerability Analysis in Patients with Relapsing Remitting and Primary Progressive Multiple Sclerosis in Southeastern Ontario (P2-6.014)

A Novel Hole System Dimension Analysis Tool Based on the Combination of the Process Path and Design Path

In the calculation of process dimensions in the machining of porous parts, the traditional method is complicated in the establishment of the dimensional chain, it is easy to make mistakes in the judgment of increasing and decreasing loops, and there is difficulty in the calculation process; thus, this paper puts forward a method that combines the process path with the design path to analyze the hole system dimensions in response to these problems. First of all, the combination of the process path and design path is used to establish a system of vector equations and, according to the system of vector equations, to establish a matrix model to calculate the process dimensions based on the system of vector equations to construct the process dimensional chain and the dimensional chain function; then, the principle of differentiation is used to carry out a tolerance analysis of the process dimensions, which avoids the drawing of dimensional chain diagrams and the judgement of the incremental/decremental loops. Finally, the development of a process dimension analysis tool for hole system dimensions is completed based on CATIA CAA. In order to verify the effectiveness of the proposed method, the analysis tool was used to solve the three labeling methods of the hole system dimensions of porous parts and compared with the results calculated using the traditional method, and it was found that some of the tolerance results obtained by the proposed method were enlarged by about 50%, which is more in line with the actual production situation. Finally, in order to verify the accuracy of the tool’s calculation results, three porous parts were machined according to the process dimensions and tolerances calculated by the tool, and the center distance of each hole was measured by a coordinate meter. These measurements were compared and analyzed with the design dimensions specified in the engineering drawings, and it was found that the center distances of the holes were in full compliance with the design requirements.

Comprehensive performance domain tolerance analysis methodology for freeform imaging spectrometers

In recent years, attention has been directed towards cost-effective and compact freeform Schwarzschild imaging spectrometers with plane gratings. The utilization of tolerance analysis serves as a potent approach to facilitate the development of prototypes. Conventional tolerance analysis methods often rely solely on the modulation transfer function (MTF) criterion. However, for a spectrometer system, factors such as the keystone/smile distortion and spectral resolution performance also require consideration. In this study, a tailored comprehensive performance domain tolerance analysis methodology for freeform imaging spectrometers was developed, considering vital aspects such as the MTF, keystone/smile distortion, and spectral resolution. Through this approach, meticulous tolerance analysis was conducted for a freeform Schwarzschild imaging spectrometer, providing valuable insights for the prototype machining and assembly processes. Emphasis was placed on the necessity of precise control over the tilt and decenter between the first and third mirrors, whereas the other fabrication and assembly tolerances adhered to the standard requirements. Finally, an alignment computer-generated hologram (CGH) was employed for the preassembly of the first and third mirrors, enabling successful prototype development. The congruence observed between the measured results and tolerance analysis outcomes demonstrates the effectiveness of the proposed method.

Analysis and Optimization of Error Tolerance in an Incoherent Beam Combination System

Analysis and Optimization of Error Tolerance in an Incoherent Beam Combination System

Carbohydrase activity in proximal and distal segments of small intestine in health, irritated bowel syndrome and enteropathy with membrane digestion disturbance

Diagnostics of enteropathy with membrane digestion disturbance (EMDD) is based on the evaluation of carbohydrase activity in duodenal mucosa (DM). There is no information about diagnostic value of ileum mucosa (IM) carbohydrases.
 Aim. Assess the diagnostic value of carbohydrases activity in mucous membrane of duodenum and distal ileum in patients with EMDD and irritable bowel syndrome (IBS).
 Materials and methods. The study includes 12 patients with EMDD, 16 patients with IBS and 12 relatively healthy persons (control group). Study design included analysis of food tolerance and evaluation of activity of sucrase, glucoamylase, maltase and lactase in mucous membrane of duodenum and ileum based on the method of Dahlquist with N.I. Belostotsky modification. Microsoft Office Excel and Statistica 10.0 were used for statistical evaluation of material. Mann–Whitney criterion was used to evaluate the differences between quantitative parameters. The differences were considered to be significant at p0.05. Control group was used for comparison.
 Results. In control group the carbohydrases activity in duodenum mucosa was not significantly statistically different from the one in IM. In patients of the IBS group, the activity of glucoamylase and maltase in the IM was significantly higher than in the duodenum: glucoamylase by 2.2 times, maltase by 2 times. In patients with EMDD the activity of glucoamylase, maltase and sucrase in IM is higher than in DM by 5.2, 3.8 and 2 times respectively, which signifies the formation of compensatory reaction of adaptive shift, initially described by A.M. Ugolev in 1985.
 Conclusion. The evaluation of carbohydrase activity in IM provides more information about pathogenesis of EMDD and its differential diagnosis with IBS.

Research on the Calculation Method for the Load Distribution of the Dual-Input Counter-Rotating Transmission System

In order to solve the problem of an uneven load distribution in a double-input and double-output gearbox, based on the small-displacement Jacobi spin theory and considering the manufacturing and assembly errors of the gearbox, a Jacobi spin analysis model of the gearbox was established. In order to improve the accuracy of the three-dimensional tolerance analysis of the gearbox, on the basis of the processing method of parallel chain of gears and shafts and box and shafts, the serialization method of parallel chain of meshing gear pairs is proposed, and the gearbox was established. For the three-dimensional tolerance analysis model, three-dimensional tolerance analysis was carried out on the side-clearance dimensions of double-input and double-output gearboxes, and the side clearance values under the extreme value method and Monte Carlo method were deduced and combined with the theoretical side clearance to obtain the range of the side clearance of the gearboxes. Elastic deflection analysis was carried out on the gearbox shaft system, and the compensation relationship between the elastic deflection and the side clearance was investigated, so as to calculate the range of the value of the gearbox uniform load coefficient from 1 to 1.259. The error between the theoretically calculated uniform load coefficient and the experimentally tested uniform load coefficient was 6.61%, which verifies the reasonableness of the load distribution method of double-input and double-output gearboxes calculated using three-dimensional tolerance analysis combined with the elastic deflection angle, and provides a theoretical basis for the optimal design of the uniform load of multi-branch input gearboxes.

OsEIN2-OsEIL1/2 pathway negatively regulates chilling tolerance by attenuating OsICE1 function in rice.

Low temperature severely affects rice development and yield. Ethylene signal is essential for plant development and stress response. Here, we reported that the OsEIN2-OsEIL1/2 pathway reduced OsICE1-dependent chilling tolerance in rice. The overexpressing plants of OsEIN2, OsEIL1 and OsEIL2 exhibited severe stress symptoms with excessive reactive oxygen species (ROS) accumulation under chilling, while the mutants (osein2 and oseil1) and OsEIL2-RNA interference plants (OsEIL2-Ri) showed the enhanced chilling tolerance. We validated that OsEIL1 and OsEIL2 could form a heterxodimer and synergistically repressed OsICE1 expression by binding to its promoter. The expression of OsICE1 target genes, ROS scavenging- and photosynthesis-related genes were downregulated by OsEIN2 and OsEIL1/2, which were activated by OsICE1, suggesting that OsEIN2-OsEIL1/2 pathway might mediate ROS accumulation and photosynthetic capacity under chilling by attenuating OsICE1 function. Moreover, the association analysis of the seedling chilling tolerance with the haplotype showed that the lower expression of OsEIL1 and OsEIL2 caused by natural variation might confer chilling tolerance on rice seedlings. Finally, we generated OsEIL2-edited rice with an enhanced chilling tolerance. Taken together, our findings reveal a possible mechanism integrating OsEIN2-OsEIL1/2 pathway with OsICE1-dependent cascade in regulating chilling tolerance, providing a practical strategy for breeding chilling-tolerant rice.

Integrated analysis of transcriptomic and physiological responses to cold stress in Macrobrachium rosenbergii

The giant freshwater prawn (GFP), Macrobrachium rosenbergii, stands out as a vital species in freshwater shrimp aquaculture. However, its susceptibility to cold conditions has increased the cost of aquaculture and limited the growth of GFP cultivation. To furnish insights at the molecular level to enhance GFP's cold tolerance, transcriptome sequencing and physiological analysis of the hepatopancreas were performed in our study. Initially, we identified 4909 differentially expressed genes (DEGs) between experimental (15 °C, 24 h) and control groups (25 °C, 24 h). These DEGs were significantly enriched in the proteasome, spliceosome, lysosome, peroxisome, and lipid metabolism-related pathways including glycerolipid metabolism, sphingolipid metabolism, linoleic acid metabolism as well as steroid hormone biosynthesis. Histological examination following various stress durations revealed a notable increase in hepatocyte vacuoles after 6 h of cold stress. Hepatocyte structure deterioration occurred in the later phase (48 h), coinciding with a significant increase in cell apoptosis. Subsequent analyses demonstrated an initial increase followed by a decrease in superoxide dismutase (SOD), catalase (CAT), and glutathione (GSH) content, while malonaldehyde (MDA) content rose with stress duration. Furthermore, a significant increase in triglyceride content and no significant change in cholesterol content was observed. RT-qPCR results unveiled significant down-regulated expression of FAS, TAGL, ACOX1, ACS and significant up-regulated expression of GPAT and DEGS. These findings suggest that oxidative stress and impaired lipid metabolism underlie the poor cold tolerance of GFP.

Вплив зрошення мінералізованими водами на процеси засолення ґрунтів Південно-Бузької та Кам’янської зрошувальних систем

In Ukraine, irrigation development is hampered by a shortage of high-quality irrigation water. Irrigation with mineralized irrigation water causes soil salinization, which negatively affects crop productivity, soil quality, and surface water conditions. The purpose of this article is to analyze the impact of irrigation water of different mineralization on soil salinity in the Southern Bug and Kamianske irrigation systems. The field and laboratory studies used the methods and criteria of the salinity laboratory of the US Department of Agriculture and the Food and Agriculture Organization. The studies showed that irrigation is carried out with water that has an average risk of soil salinity (ECw = 0.7-3.0 dS/m). At the same time, there is some spatial differentiation in the quality of irrigation water. The lands of the Southern Bug irrigation system are irrigated with water with an electrical conductivity of 0.90-1.07 dS/m, and the lands of the Kamianska irrigation system are irrigated with water with an electrical conductivity of 1.86-2.65 dS/m. The mineralization of irrigation water determines the degree of soil salinity. While the soils of the Southern Bug irrigation system are not saline (ECe is less than 2.0 dS/m), the soils of the Kamianske irrigation system have a significant accumulation of salts (ECe reaches 5.5 dS/m). According to the Food and Agriculture Organization classification, such soils are classified as saline. Analysis of the salinity tolerance of individual crops showed that any crops of any salt tolerance can be grown on the irrigated lands of the Southern Bug irrigation system. And on the lands of the Kamianske irrigation system, only moderately sensitive and least sensitive crops to soil salinity can be grown without loss of yield. The conducted research allows us to adjust the structure of sown areas and the structure of crop rotations in terms of salt tolerance of certain crops.

Isolation and screening of stress tolerant and plant growth promoting root nodulating rhizobial bacteria from some wild legumes of Nagaland, India

Wild legumes are widely distributed and better adapted to extreme agro-climatic conditions because of their soil microbiota. In the present study, Root Nodulating Bacteria (RNB) were isolated from wild legumes and analysed for their abiotic stress tolerant and Plant Growth Promoting Traits (PGPT). Rhizobial strains were identified based on their 16S rRNA sequences. A total of 14 rhizobial species were characterized based, of which six were Rhizobium (AIS3, AIS9, AIR14, LUMCR3, LUMVRW4, LUMVRW8), four belongs to Burkholderia (AIS12, MOLKCS15, CHMP9, CHMP10) and one each of genera Cupriavidus (LUMMD12), Herbaspirillum (LUMVRW4), Paraburkholderia (CHMP1) and Ralstonia (LUMDes9). Stress tolerance analysis found most of the strains are to be tolerant to alkaline pH (9–11) where highest tolerance exhibited by Rhizobium sp. (strain AIS9) and Burkholderia territorii (strain MOKCS15). Seven Rhizobial strains were able to thrive at lower temperature stress (10 and 20 °C); while, strains AIS9, CHMP1 and LUMMD21 were able to grow at higher temperature (40 and 50 °C). Most of the isolate could tolerate salinity stress up to 1.5 %; but, only isolates AIS9, AIS12 and LUMMD12 could tolerate 2 and 2.5 % salt concentrations. The PGPT analyses found LUMCR3 having the highest IAA production potentiality (102.5 µg/ml); while, isolate MOKCS15 exhibited highest phosphate solubilizing activity (4.1 Phosphate Solubilization Index). The RNB isolates in the present study exhibited both stress tolerance and PGP traits indicating their potential use as biofertilizer under extreme agronomic conditions to improve productivity.

Riboswitch-controlled IL-12 gene therapy reduces hepatocellular cancer in mice.

Hepatocellular carcinoma (HCC) and solid cancers with liver metastases are indications with high unmet medical need. Interleukin-12 (IL-12) is a proinflammatory cytokine with substantial anti-tumor properties, but its therapeutic potential has not been realized due to severe toxicity. Here, we show that orthotopic liver tumors in mice can be treated by targeting hepatocytes via systemic delivery of adeno-associated virus (AAV) vectors carrying the murine IL-12 gene. Controlled cytokine production was achieved in vivo by using the tetracycline-inducible K19 riboswitch. AAV-mediated expression of IL-12 led to STAT4 phosphorylation, interferon-γ (IFNγ) production, infiltration of T cells and, ultimately, tumor regression. By detailed analyses of efficacy and tolerability in healthy and tumor-bearing animals, we could define a safe and efficacious vector dose. As a potential clinical candidate, we characterized vectors carrying the human IL-12 (huIL-12) gene. In mice, bioactive human IL-12 was expressed in a vector dose-dependent manner and could be induced by tetracycline, suggesting tissue-specific AAV vectors with riboswitch-controlled expression of highly potent proinflammatory cytokines as an attractive approach for vector-based cancer immunotherapy.