Performance of spleen stiffness measurement to rule out high-risk varices in patients with porto-sinusoidal vascular disorder.

BACKGROUND AIMS: Baveno VII consensus suggests that screening endoscopy can be spared in patients with compensated cirrhosis when spleen stiffness measurement (SSM) by vibration-controlled transient elastography (VCTE) is ≤40 kPa as they have a low probability of high-risk varices (HRV). Conversely, screening endoscopy is required in all patients with porto-sinusoidal vascular disorder (PSVD). This study aimed to evaluate the performance of SSM-VCTE to rule out HRV in patients with PSVD and signs of portal hypertension. APPROACH RESULTS: We retrospectively included patients with PSVD, ≥1 sign of portal hypertension, without history of variceal bleeding, who underwent a SSM-VCTE within 2 years before or after an upper endoscopy in 21 VALDIG centers, divided into a derivation and a validation cohort. 154 patients were included in the derivation cohort; 43% had HRV. By multivariable logistic regression analysis, SSM-VCTE >40 kPa and serum bilirubin ≥1 mg/dL were associated with HRV. SSM-VCTE ≤40 kPa combined with bilirubin <1 mg/dL had a sensitivity of 96% to rule out HRV, and could spare 38% of screening endoscopies, with 4% of HRV missed, and a 95% negative predictive value (NPV). In the validation cohort, including 155 patients, SSM combined with bilirubin could spare 21% of screening endoscopies, with 4% of HRV missed and a 94% NPV. CONCLUSION: This study gathering a total of 309 PSVD patients showed that SSM-VCTE ≤40 kPa combined with bilirubin <1 mg/dL identifies patients with PSVD and portal hypertension with a probability of HRV <5%, in whom screening endoscopy can be spared.

Reproducibility and accuracy of a pocket-size ultrasound device in assessing liver steatosis.

BACKGROUND: This diagnostic prospective study compared the feasibility and diagnostic accuracy of Pocket-size Ultrasound Devices (PUDs) against standard ultrasound (US) in detecting liver steatosis using the controlled attenuation parameter (CAP) and liver biopsy as reference standards. MATERIALS AND METHODS: Consecutive patients with chronic liver diseases were assessed for the presence of steatosis using PUD and US. A CAP cut-off value >275 dB/m was applied to establish ≥S1. A 26-patient subgroup underwent liver biopsy. PUD reproducibility was evaluated using Cohen's k statistic. Diagnostic accuracy of PUD and US was given as Sensibility (Sn), Specificity (Sp), Positive and Negative Predictive Values (PPV, NPV), positive and negative Likelihood Ratio (LR+, LR-). RESULTS: 81 consecutive patients (69% males) with multiple etiologies were enroled. PUD inter-observer agreement was good (k 0.77, 95%CI 0.62-0.93). PUD and US identified ≥S1 according to CAP values respectively with Sn 0.87, Sp 0.61, PPV 0.49, NPV 0.91, LR+ 2.04, LR- 0.07, AUROC 0.74 and Sn 0.96, Sp 0.54, PPV 0.47, NPV 0.97, LR+ 2.10, LR- 0.07, AUROC 0.75. CONCLUSIONS: PUD shows good reproducibility and diagnostic accuracy in ruling liver steatosis out, representing a useful point-of-care tool to avail of hepatologists interested in excluding NAFLD, but with basic US skills.

Spike-specific humoral and cellular immune responses after COVID-19 mRNA vaccination in patients with cirrhosis: A prospective single center study.

BACKGROUND AND AIMS: COVID-19 mRNA vaccines were approved to prevent severe forms of the disease, but their immunogenicity and safety in cirrhosis is poorly known. METHOD: In this prospective single-center study enrolling patients with cirrhosis undergoing COVID-19 vaccination (BNT162b2 and mRNA-1273), we assessed humoral and cellular responses vs healthy controls, the incidence of breakthrough infections and adverse events (AEs). Antibodies against spike- and nucleocapsid-protein (anti-S and anti-N) and Spike-specific T-cells responses were quantified at baseline, 21 days after the first and second doses and during follow-up. RESULTS: 182 cirrhotics (85% SARS-CoV-2-naïve) and 38 controls were enrolled. After 2 doses of vaccine, anti-S titres were significantly lower in cirrhotics vs controls [1,751 (0.4-25,000) U/mL vs 4,523 (259-25,000) U/mL, p=0.012] and in SARS-CoV-2-naïve vs previously infected cirrhotics [999 (0.4-17,329) U/mL vs 7,500 (12.5-25,000) U/mL, (p<0.001)]. T-cell responses in cirrhotics were similar to controls, although with different kinetics. In SARS-CoV-2-naïve cirrhotics, HCC, Child-Pugh B/C and BNT162b2 were independent predictors of low response. Neither unexpected nor severe AEs emerged. During follow-up, 2% turned SARS-CoV-2 positive, all asymptomatic. CONCLUSION: Humoral response to COVID-19 vaccines appeared suboptimal in patients with cirrhosis, particularly in SARS-CoV-2-naïve decompensated cirrhotics, although cellular response appeared preserved, and low breakthrough infections rate was registered.

Jump Rope Training: Balance and Motor Coordination in Preadolescent Soccer Players.

General physical practice and multidimensional exercises are essential elements that allow young athletes to enhance their coordinative traits, balance, and strength and power levels, which are linked to the learning soccer-specific skills. Jumping rope is a widely-used and non-specific practical method for the development of athletic conditioning, balance and coordination in several disciplines. Thus, the aim of this study was to investigate the effects of a short-term training protocol including jumping rope (JR) exercises on motor abilities and body balance in young soccer players. Twenty-four preadolescent soccer players were recruited and placed in two different groups. In the Experimental group (EG), children performed JR training at the beginning of the training session. The control group (CG), executed soccer specific drills. Harre circuit test (HCT) and Lower Quarter Y balance test (YBT-LQ) were selected to evaluate participant's motor ability (e.g. ability to perform rapidly a course with different physical tasks such as somersault and passages above/below obstacles ) and to assess unilateral dynamic lower limb balance after 8 weeks of training. Statistical analysis consisted of paired t-test and mixed analysis of variance scores to determine any significant interactions. Children who performed jumping rope exercises showed a significant decrease of 9% (p < 0.01, ES = 0.50-0.80) in the performance time of HCT. With regard to the CG, no differences were highlighted (p > 0.05, ES = 0.05-0.2) from pre- to post-training. A training-by-group interaction was found for the composite score in both legs (p < 0.05, Part η(2) > 0.14). Our findings demonstrated that JR practice within regular soccer training enhanced general motor coordination and balance in preadolescent soccer players. Therefore, the inclusion of JR practice within regular soccer training session should encouraged to improve children's motor skills. Key pointsPerforming jumping rope exercises within a regular soccer program can be an additional method to improve balance and motor coordination.The performance improvement in the Harre Circuit Test associated with jump rope training can potentially be attributed to an enhancement of the inter-limb coordination and SSC ability.Results from the present study indicate that young soccer players should be encouraged to practice general physical activities together with sport-specific exercise during their training sessions.

Constitutive co-suppression of the GA 20-oxidase1 gene in tomato leads to severe defects in vegetative and reproductive development.

To dissect the role of gibberellins in tomato development, we have constitutively down-regulated the gene GA 20-oxidase1 (GA20ox1). Plants co-suppressed for GA20ox1 (referred to as CO-6 plants) showed vegetative defects typical of GA deficiency such as darker and mis-shaped leaves and dwarfism. CO-6 plants flowered as the controls, although their flowers had subtle defects in the pedicel and in organ insertion. Analysis of male development revealed defects before, during and after meiosis, and a final pollen viability of 22%. The development of female organs and gametes appeared normal. Pollination experiments indicated that the pollen produced by CO-6 plants was able to fertilize control ovaries, but the analysis of the progeny showed that the construct was not transmitted. Ovaries of CO-6 plants showed high fruit set and normal fruit development when pollinated with control pollen. However these fruits were completely seedless due to a stenospermocarpic behaviour that was evidenced by callose layering in the endothelium between 7 and 15 days after pollination. We conclude that GA20ox1 in tomato exerts specific developmental roles that are not redundantly shared with other members of this gene family. For reproductive male development, silencing of this gene is detrimental for pollen production and either gametophytically lethal or severely hampering seed germination. In the pistil, the co-suppression construct does not affect the progamic phase, nor fruit set and growth, but it interferes with seed development after fertilization leading to seed abortion.

Tomato fruit set driven by pollination or by the parthenocarpic fruit allele are mediated by transcriptionally regulated gibberellin biosynthesis.

We investigated the role of gibberellins (GAs) in the phenotype of parthenocarpic fruit (pat), a recessive mutation conferring parthenocarpy in tomato (Solanum lycopersicum L.). Novel phenotypes that parallel those reported in plants repeatedly treated with gibberellic acid or having a GA-constitutive response indicate that the pat mutant probably expresses high levels of GA. The retained sensitivity to the GA-biosynthesis inhibitor paclobutrazol reveals that this condition is dependent on GA biosynthesis. Expression analysis of genes encoding key enzymes involved in GA biosynthesis shows that in normal tomato ovaries, the GA20ox1 transcript is in low copy number before anthesis and only pollination and fertilization increase its transcription levels and, thus, GA biosynthesis. In the unpollinated ovaries of the pat mutant, this mechanism is de-regulated and GA20ox1 is constitutively expressed, indicating that a high GA concentration could play a part in the parthenocarpic phenotype. The levels of endogenous GAs measured in the floral organs of the pat mutant support such a hypothesis. Collectively, the data indicate that transcriptional regulation of GA20ox1 mediates pollination-induced fruit set in tomato and that parthenocarpy in pat results from the mis-regulation of this mechanism. As genes involved in the control of GA synthesis (LeT6, LeT12 and LeCUC2) and response (SPY) are also altered in the pat ovary, it is suggested that the pat mutation affects a regulatory gene located upstream of the control of fruit set exerted by GAs.

A H2O2-forming peroxidase rather than a NAD(P)H-dependent O2•- synthase may be the major player in cell death responses controlled by the Pto-Fen complex following fenthion treatment.

Four tomato (Lycopersicon esculentum Mill.) near-isogenic lines were treated by foliar spraying with the insecticide fenthion. Two, Riogrande and Rimone, differed from each other only for the presence in the latter of the Fen gene, conferring propensity to develop foliar symptoms upon exposure to fenthion. The other two, namely RC332 and RC131, were the transgenic versions of Riogrande and Rimone, respectively, harbouring the Gox gene encoding for glucose oxidase of Aspergillus niger. The production of H2O2 as well as the activities of H+-ATPase, NAD(P)H-dependent superoxide synthase, catalase, peroxidase, and Cu,Zn-superoxide dismutase were evaluated in the foliar tissues up to 24 h after exposure to fenthion. The Fen gene conferred sensitivity to fenthion, regardless of the expression of a Gox transgene. A prolonged accumulation of H2O2 was observed in the leaves of Rimone and of RC131, which was instead transient in Riogrande and in RC332. In all the tomato lines, exposure to fenthion induced rapid but transient changes in the activities of most enzymes. The only exception was peroxidase activity in the leaves of Rimone and of RC131, which steadily increased until the end of the sampling period. It is suggested that the sensitivity of Rimone to fenthion might be due to the sustained activity of a H2O2-forming peroxidase.

The Cf-2 / Rcr3esc gene interaction in tomato (Lycopersicon esculentum) induces autonecrosis and triggers biochemical markers of oxidative burst at cellular level.

A tomato plant (Lycopersicon esculentum Mill.) with necrotic leaf spots mimicking disease lesions was singled out in progeny under selection in Moscow (breeding material of Ignatova Svetlana). The progeny from spontaneous selfing of such a plant (V20368), in the presence of increasing temperature and high light intensity, exhibited spontaneous necrotic lesions on the leaves, with acropetal progression (autonecrosis). A similar phenotype, described in 1948 by Langford, appeared to be associated with the Cf-2 resistance gene, introgressed from L. pimpinellifolium. Recently, Kruger et al. (2002) demonstrated that the Cf-2 effect depends on a second gene (Rcr3pim) encoding a cysteine protease, and that autonecrosis is activated by the contemporary presence of Cf-2 and the L. esculentum allele Rcr3esc. In this work we characterised the V20368 mutant and verified that autonecrosis is caused by the presence of an interaction between Cf-2 and Rcr3esc. When the environmental conditions are favourable, this interaction triggers an oxidative burst, as evidenced by a strong increase in H2O2 production and activities of catalase (EC 1.11.1.6), peroxidase (EC 1.11.1.7) and ATPase (EC 3.6.1.3). In addition, by grafting the necrotic mutant on the cv. Riogrande and vice versa, we proved that the necrotic phenotype is not associated with the movement of a signal molecule, since the autonecrosis was not transmitted across the grafting point. Finally, the interaction between Cf-2 and Rcr3esc appeared to lower the threshold of stress perception, as evidenced by an increased sensitivity to the insecticide Fenthion.

Tomato transgenic lines and Tetranychus urticae: changes in plant suitability and susceptibility.

A critical aspect dealing with the use of transgenic plants is the global evaluation of their environmental impact. The polyphagous mite Tetranychus urticae can be considered a suitable species to investigate unpredictable and undesirable effects on phytophagous arthropods. Three tomato near isogenic lines, that is, the cv. Riogrande (RIG), the transgenic lines RC332 (containing the Gox gene and showing high glucose oxidase activity), and MS498 (containing the KTI3 gene and exhibiting a high trypsin inhibition) were used in laboratory and greenhouse trials. Trichomes and contents of C and N of the leaves, differences in development and oviposition of T. urticae and damage caused were evaluated for each line. The laboratory trials evidenced that (1) the intrinsic rate of increase of two strains of T. urticae (T from tomato, B from bindweed), reared on the lower surface of tomato leaflets, was significantly lower in RIG than in transgenic lines and doubling time ranged between 6.9 and 11.6 days in the first and between 3.9 and 5.3 days in the latter; (2) the glandular four-lobed trichomes were always higher in RIG than in other genotypes; (3) the N leaf content was from 1.3 to 1.9 fold lower and the C/N ratio from 1.3 to 1.9 fold higher in RIG than in other lines. The greenhouse experiment, that lasted over a month and was performed by inducing an initially equal infestation of strain T, evidenced: (1) no significant difference between plant lines in the final mite infestation (motile stages per plant), nevertheless an almost double number of spider mites was counted in RC332; (2) a significantly higher percentage of damaged leaves and a significant higher average damage index on RC332 than on RIG (79% and 2.3 in the former, and 62% and 2.1 in the latter, respectively), even if in both transgenics a higher level of the most severe damages and a shorter time to approach them were observed; (4) a comparable number of mites causing the same damage level in all genotypes and a strong linear relation between the first four levels of damage and mite infestation. Although in the laboratory studies both transgenic lines enhanced the T. urticae population increase, the glasshouse studies were not as conclusive and they only suggest the possibility of any real difference between the transgenic and non-transgenic genotypes.