Sulfate is Incorporated into Cysteine to Trigger ABA Production and Stomatal Closure.

Plants close stomata when root water availability becomes limiting. Recent studies have demonstrated that soil-drying induces root-to-shoot sulfate transport via the xylem and that sulfate closes stomata. Here we provide evidence for a physiologically relevant signaling pathway that underlies sulfate-induced stomatal closure in Arabidopsis (Arabidopsis thaliana). We uncovered that, in the guard cells, sulfate activates NADPH oxidases to produce reactive oxygen species (ROS) and that this ROS induction is essential for sulfate-induced stomata closure. In line with the function of ROS as the second-messenger of abscisic acid (ABA) signaling, sulfate does not induce ROS in the ABA-synthesis mutant, aba3-1, and sulfate-induced ROS were ineffective at closing stomata in the ABA-insensitive mutant abi2-1 and a SLOW ANION CHANNEL1 loss-of-function mutant. We provided direct evidence for sulfate-induced accumulation of ABA in the cytosol of guard cells by application of the ABAleon2.1 ABA sensor, the ABA signaling reporter ProRAB18:GFP, and quantification of endogenous ABA marker genes. In concordance with previous studies, showing that ABA DEFICIENT3 uses Cys as the substrate for activation of the ABSCISIC ALDEHYDE OXIDASE3 (AAO3) enzyme catalyzing the last step of ABA production, we demonstrated that assimilation of sulfate into Cys is necessary for sulfate-induced stomatal closure and that sulfate-feeding or Cys-feeding induces transcription of NINE-CIS-EPOXYCAROTENOID DIOXYGENASE3, limiting the synthesis of the AAO3 substrate. Consequently, Cys synthesis-depleted mutants are sensitive to soil-drying due to enhanced water loss. Our data demonstrate that sulfate is incorporated into Cys and tunes ABA biosynthesis in leaves, promoting stomatal closure, and that this mechanism contributes to the physiological water limitation response.

SARS-CoV-2: big seroprevalence data from Pakistan-is herd immunity at hand?

PURPOSE: Seroprevalence surveys from different countries have reported SARS CoV-2 antibodies below 20% even in the most adversely affected areas and herd immunity cannot be predicted till more than half of the population gets the disease. The purpose of this survey was to estimate the magnitude of community-based spread of the infection, associated immunity, and the future prospects and proximity to a 'herd community'. METHODS: The study was undertaken as a cluster randomized, cross-sectional countrywide survey. This largest community-based seroprevalence data of SARS-CoV-2 were collected between 15th and 31st July, 2020 from seven randomly selected cities belonging to the three most populous provinces of Pakistan. The FDA approved kit of ROCHE was used for detection of SARS-CoV-2 antibodies. RESULTS: Serum samples of 15,390 participants were tested for SARS CoV-2 antibodies with an overall seroprevalence of 42.4%. The seroprevalence ranged from 31.1% to 48.1% in different cities with the highest in Punjab province (44.5%). In univariable analysis, the odds of seropositivity was higher in men compared to women (OR: 1.10, 95% CI: 1.01-1.19, P < 0.05). In multivariable analysis, the risk of being seropositive was lower (OR 0.72, 95% CI: 0.60-0.87, P < 0.01) in younger group (≤ 20 years) than in those aged above 60 years. CONCLUSION: The study concluded that despite a reasonable seroprevalence, the country is yet to reach the base minimum of estimations for herd immunity. The durability of immunity though debated at the moment, has shown an evidenced informed shift towards longer side.

Incidence of SARS-CoV-2 re-infection in anti-nucleocapsid IgG-positive healthcare workers: a prospective cohort study.

BACKGROUND : Since the pandemic of SARS-CoV-2 began, our understanding of the pathogenesis and immune responses to this virus has continued to evolve. It has been shown that this infection produces natural detectable immune responses in many cases. However, the duration and durability of immunity and its effect on the severity of the illness are still under investigation. Moreover, the protective effects of antibodies against new SARS-CoV-2 variants still remain unclear. OBJECTIVES: To assess the incidence and associated demographic features of SARS-CoV-2 infection in anti-nucleocapsid IgG-positive and anti-nucleocapsid IgG-negative healthcare workers. MATERIAL AND METHODS: This prospective longitudinal cohort study was conducted in Peshawar Medical College group of hospitals of Prime Foundation. Anti-nucleocapsid IgG sero-positive and anti-nucleocapsid IgG sero-negative healthcare workers were followed for a period of 6 months (from 1 Aug 2020 to 31 Jan 2021), and the incidence of SARS-CoV-2 was confirmed by RT-PCR. RESULTS: A total number of 555 cohorts were followed for a period of 6 months; of them 365 (65.7%) were anti-nucleocapsid-negative (group A) and 190 (34.3%) were anti-nucleocapsid-positive (group B) healthcare workers. The mean age of the study cohort was 33.85 ± 9.80 (anti-N (-), 34.2 ± 10.58; anti-N ( +), 33.5 ± 9.50). The median antibody level in anti-nucleocapsid-positive HCWs was 15.95 (IQR: 5.24-53.4). Male gender was the majority in both groups (group A, 246 (67%), group B, 143 (48%)) with statistically significant difference (P < 0.05). Majority of the HCWs were blood group B in both groups (34% each). None of the 190 anti-nucleocapsid-positive HCWs developed subsequent SARS-CoV-2 re-infection, while 17% (n = 65) HCWs developed infection in anti-nucleocapsid-negative group during the 6-month follow-up period. CONCLUSION: In conclusion, none of the anti-nucleocapsid-positive HCWs developed SARS-CoV-2 re-infection in this study, and the presence of IgG anti-nucleocapsid antibodies substantially reduce the risk of re-infection for a period of 6 months.

Crosstalk Between Iron and Sulfur Homeostasis Networks in Arabidopsis.

The widespread deficiency of iron (Fe) and sulfur (S) is becoming a global concern. The underlying mechanisms regulating Fe and S sensing and signaling have not been well understood. We investigated the crosstalk between Fe and S using mutants impaired in Fe homeostasis, sulfate assimilation, and glutathione (GSH) biosynthesis. We showed that chlorosis symptoms induced by Fe deficiency were not directly related to the endogenous GSH levels. We found dynamic crosstalk between Fe and S networks and more interestingly observed that the upregulated expression of IRT1 and FRO2 under S deficiency in Col-0 was missing in the cad2-1 mutant background, which suggests that under S deficiency, the expression of IRT1 and FRO2 was directly or indirectly dependent on GSH. Interestingly, the bottleneck in sulfite reduction led to a constitutively higher IRT1 expression in the sir1-1 mutant. While the high-affinity sulfate transporter (Sultr1;2) was upregulated under Fe deficiency in the roots, the low-affinity sulfate transporters (Sultr2;1, and Sultr2;2) were down-regulated in the shoots of Col-0 seedlings. Moreover, the expression analysis of some of the key players in the Fe-S cluster assembly revealed that the expression of the so-called Fe donor in mitochondria (AtFH) and S mobilizer of group II cysteine desulfurase in plastids (AtNFS2) were upregulated under Fe deficiency in Col-0. Our qPCR data and ChIP-qPCR experiments suggested that the expression of AtFH is likely under the transcriptional regulation of the central transcription factor FIT.

Identifying higher risk subgroups of health care workers for priority vaccination against COVID-19.

Background: Health care workers (HCWs) are exposed to high risk of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection due to close contact with infected patients in hospital. The objective of this study was to estimate the seroprevalence and to identify the exposure risk of various subgroups among HCWs to prioritize them for early vaccination. Methods: This was a multicentre cross-sectional study conducted between 15 and 29 June 2020. A total of 987 HCWs were recruited randomly from two major tertiary-care hospitals of Peshawar city, Pakistan. The HCWs included doctors, nurses, paramedics and hospital support staff. The US Food and Drug Administration (FDA)-approved kit was used for the detection of SARS-CoV-2 antibodies. Results: Overall, 310 (31.4%) HCWs were seropositive for SARS-CoV-2 antibodies (95% confidence interval, CI: 28.5-34.4). Seroprevalence was higher in males (33.5%) and in age group 51-60 years (40.9%). Seropositivity increased with increasing age from 8.3% in age group ⩽20 to 40.9% in 51-60 years of age group (p < 0.05). The highest seroprevalence was identified in paramedical staff (42·5%, 95% CI: 36.6-48.6) followed by nursing staff (38·8%, 95% CI: 32.1-45.7). In logistic regression, being a male HCW led to higher risk of seropositivity (odds ratio, OR: 1.50, 95% CI: 1·06-2.13. p < 0.05) compared with female staff members. The odds of seropositivity was higher in nurses (OR: 3·47, 95% CI: 1.99-6.05. p < 0.01), paramedical staff (OR: 3·19, 95% CI: 1.93-5.28. p < 0.01) and hospital support staff (OR: 2·47, 95% CI: 1.29-4.7. p < 0.01) compared with consultants. Conclusion: Overall, our results concluded that nursing and paramedical staff are at higher risk and should be vaccinated on priority.

Sulfur metabolic engineering enhances cadmium stress tolerance and root to shoot iron translocation in Brassica napus L.

Serine acetyltransferase (SAT) (EC 2.3.1.30) is the rate-limiting enzyme of cysteine (Cys) biosynthesis, providing the decisive precursor for the ubiquitous defense thiol glutathione (GSH). Together with O-acetylserine (thiol) lyase (OAS-TL; EC 2.5.1.47) SAT generates Cys in the cytosol, plastids, and mitochondria of vascular plants. The current study aimed to overproduce Cys and GSH for enhanced stress tolerance via overexpression of the feedback-insensitive isoform of serine acetyltransferase from tobacco, i.e., NtSAT4. Constitutive overexpression of NtSAT4 in Brassica napus resulted in the 2.6-fold-4-fold higher SAT activity in different subcellular compartment-specific lines. This higher SAT activity led to a 2.5-fold-3.5-fold higher steady-state level of free Cys and 2.2-fold-5.3-fold elevated level of GSH in leaves compared with nontransformed plants. Among the compartment-specific lines, the mitochondrial targeted NtSAT4 overexpressor line M-182 showed the highest levels of Cys (3.5-fold) and GSH (5.3-fold) compared with wild-type plants. Overexpression of NtSAT4 conferred a physiological advantage in terms of enhanced tolerance against oxidative stress with hydrogen peroxide and the heavy metal cadmium (Cd). The NtSAT4 overexpressor lines showed a significantly higher amount of iron (Fe) translocation from roots to shoots compared with nontransformed plants. Overall, these results suggest that overexpression of NtSAT4 is a promising approach to creating plants with tolerance to heavy metals and oxidative stress and, in addition, may potentially improve plant nutrition in terms of enhanced Fe translocation from roots to shoots.

Antimicrobial Prophylaxis in Clean Pediatric Surgical Procedures: A Necessity or Redundancy?

Background The clinical utility of antimicrobial prophylaxis in clean pediatric surgical cases remains enigmatic. The present study aims to evaluate the prevalence of surgical site infections in instances where antibiotic prophylaxis is not employed prior to clean pediatric surgical procedures. Methods A retrospective cross-sectional study that included data of all pediatric clean surgical procedures from January 2018 till January 2020 was conducted. All children undergoing clean surgical procedures who did not receive antibiotics at least two weeks prior to the procedure were included in the study. The exclusion criteria included patients with congenital heart disease, ventriculoperitoneal shunt, nephrotic syndrome, immunodeficiency, and prior administration of antimicrobial prophylaxis. All patients were followed for two to four weeks for any signs of surgical site infections. Results Of the 178 patients included, 119 were male and 59 were female, with the mean age hovering at 8.19 ± 2.87 years. Orchidopexy and herniotomy were the most commonly performed surgical procedures, and were performed in 56 (31.46%) and 54 (30.33%) patients, respectively. Only one case of postoperative surgical site wound infection was reported, accounting for a prevalence rate of 0.56%. Conclusion In clean pediatric surgical procedures, the risk of surgical site infections is exceedingly low. The unnecessary use of antibiotics in children can cause deleterious adverse effects and promote antimicrobial resistance. In a carefully selected pediatric population, administration of antibiotic prophylaxis might confer no added benefit.

Sulfate-Induced Stomata Closure Requires the Canonical ABA Signal Transduction Machinery.

Phytohormone abscisic acid (ABA) is the canonical trigger for stomatal closure upon abiotic stresses like drought. Soil-drying is known to facilitate root-to-shoot transport of sulfate. Remarkably, sulfate and sulfide-a downstream product of sulfate assimilation-have been independently shown to promote stomatal closure. For induction of stomatal closure, sulfate must be incorporated into cysteine, which triggers ABA biosynthesis by transcriptional activation of NCED3. Here, we apply reverse genetics to unravel if the canonical ABA signal transduction machinery is required for sulfate-induced stomata closure, and if cysteine biosynthesis is also mandatory for the induction of stomatal closure by the gasotransmitter sulfide. We provide genetic evidence for the importance of reactive oxygen species (ROS) production by the plasma membrane-localized NADPH oxidases, RBOHD, and RBOHF, during the sulfate-induced stomatal closure. In agreement with the established role of ROS as the second messenger of ABA-signaling, the SnRK2-type kinase OST1 and the protein phosphatase ABI1 are essential for sulfate-induced stomata closure. Finally, we show that sulfide fails to close stomata in a cysteine-biosynthesis depleted mutant. Our data support the hypothesis that the two mobile signals, sulfate and sulfide, induce stomatal closure by stimulating cysteine synthesis to trigger ABA production.