SifA SUMOylation governs Salmonella Typhimurium intracellular survival via modulation of lysosomal function.

One of the mechanisms shaping the pathophysiology during the infection of enteric pathogen Salmonella Typhimurium is host PTM machinery utilization by the pathogen encoded effectors. Salmonella Typhimurium (S. Tm) during infection in host cells thrives in a vacuolated compartment, Salmonella containing vacuole (SCV), which sequentially acquires host endosomal and lysosomal markers. Long tubular structures, called as Salmonella induced filaments (SIFs), are further generated by S. Tm, which are known to be required for SCV's nutrient acquisition, membrane maintenance and stability. A tightly coordinated interaction involving prominent effector SifA and various host adapters PLEKHM1, PLEKHM2 and Rab GTPases govern SCV integrity and SIF formation. Here, we report for the first time that the functional regulation of SifA is modulated by PTM SUMOylation at its 11th lysine. S. Tm expressing SUMOylation deficient lysine 11 mutants of SifA (SifAK11R) is defective in intracellular proliferation due to compromised SIF formation and enhanced lysosomal acidification. Furthermore, murine competitive index experiments reveal defective in vivo proliferation and weakened virulence of SifAK11R mutant. Concisely, our data reveal that SifAK11R mutant nearly behaves like a SifA knockout strain which impacts Rab9-MPR mediated lysosomal acidification pathway, the outcome of which culminates in reduced bacterial load in in vitro and in vivo infection model systems. Our results bring forth a novel pathogen-host crosstalk mechanism where the SUMOylation of effector SifA regulated S. Tm intracellular survival.

Coconut Water Induces Clinical Remission in Mild to Moderate Ulcerative Colitis: Double-blind Placebo-controlled Trial.

BACKGROUND & AIMS: Coconut water (CW) is anti-inflammatory, can manipulate the gut microbiome, and is a rich source of potassium. Gut microbiome modulation improves outcomes in ulcerative colitis (UC), and potassium possesses in vitro anti-inflammatory property. We evaluated the effect of CW as an adjunct therapy for patients with mild-moderate UC. METHODS: This single-center, double-blind, placebo-controlled trial randomized patients with mild to moderate (Simple Clinical Colitis Activity Index [SCCAI]: 3-9) endoscopically active UC (Ulcerative Colitis Endoscopic Index of Severity [UCEIS] >1) in 1:1 ratio to CW + standard medical therapy (SMT) vs placebo + SMT. Four hundred mL of CW was administered for 8 weeks. Primary outcome measure was clinical remission (SCCAI ≤2), and secondary outcome measures were clinical response (SCCAI decline ≥3) and adverse events at 8 weeks. Microbiome was analyzed at baseline and 8 weeks. RESULTS: Of 121 patients screened, 95 were included for modified intention to treat analysis (CW, n = 49; placebo, n = 46) (mean age, 37.2 ± 11.2 years; males, 54.1%; disease duration, 48 months [interquartile range (IQR), 24-90 months]; pancolitis, 26.1%; SCCAI, 5 [IQR, 4-6]; UCEIS, 4 [IQR, 3-5]). Clinical response (57.1% vs 28.3%; odds ratio [OR], 3.4; 95% confidence interval [CI], 1.4-7.9; P = .01), remission (53.1% vs 28.3%; OR, 2.9; 95% CI, 1.2-6.7; P = .02), and proportion of patients with fecal calprotectin (FCP) <150 µg/g (30.6% vs 6.5%; OR, 6.3; 95% CI, 1.7-23.6; P = .003) were significantly higher in CW. The relative abundance of bacterial taxa that had a significant or trend towards negative correlation with SCCAI, UCEIS, or FCP increased at 8 weeks in CW, and this effect was independent of disease activity and dietary fiber. Adverse events were comparable, and no patient developed hyperkalemia. CONCLUSIONS: CW was more effective than placebo for induction of clinical remission in patients with mild to moderate UC. The trial was prospectively registered on Clinical Trials Registry of India (ctri.nic.in, Number: CTRI/2019/03/01827).

Synthesis, Reactivity, and Complexation with Fe(0) of a Tight-bite Bis(N-heterocyclic silylene).

The synthesis, reactivity, and complexation with Fe(0) precursor of a tight-bite bis(N-heterocyclic silylene) (bis(NHSi)) ligand 1 are reported. The reaction of 1 with p-toluidine led to the activation of both N-H bonds across Si(II) atoms to afford a four-membered heterocyclic cyclodisilazane 2, with hydride substituents attached to five-coordinate Si atoms. A 1 : 2 reaction of 1 with Fe(CO)5 led to an intriguing dinuclear complex 3 featuring a five-membered (N-Si-Fe-Fe-Si) ring with a Fe-Fe bond distance of 2.6892(13) Å. All compounds (1-3) were thoroughly characterized by various spectroscopic methods and X-ray diffraction studies conclusively established their molecular structures. DFT calculations were carried out to shed light on bonding and energetic aspects in 1-3.

Modeling and tuning the electronic, mechanical and optical properties of a recently synthesized 2D polyaramid: a first principles study.

This work delves into a methodology of modeling 2D materials and their structural engineering, considering an example of a recently synthesized 2D polyaramid (2DPA-1). A bottom-up approach similar to experimental techniques is implemented for modeling, and then its electronic structures and phonon spectrum and the quadratic nature of flexural phonons are analyzed. Furthermore, boron and nitrogen atoms are substituted for the carbon atom of the amide group of 2DPA-1, and their effects on its electronic properties, phonon spectrum, and mechanical properties are compared with those of pristine 2DPA-1 using density functional theory calculations. The ab initio molecular dynamics (AIMD) simulations validate the thermal stability of our system at high temperatures. The spin-polarized electronic structures reveal the transformation of pristine 2DPA-1 from a semiconductor to a half-metal and its magnetic behaviour upon nitrogen substitution. Constraining the quadratic nature of flexural phonons using the Born-Huang criteria significantly enhances the phonon spectra, leading to more accurate and reliable simulations. For modulated 2DPA-1, the elastic modulus varies between 17 and 27 N m-1, and the absorption peaks shift from ∼5.15 eV to 2.42 eV, enabling the application of polymeric 2D nanomaterials in photocatalysis and sensing, where light absorption in the near-infrared region is important. Finally, validation of our methodology is confirmed, as computed Young's modulus (11.26-11.76 GPa) of 2DPA-1 matches excellently with the experimental value (12.7 ± 3.8 GPa). Overall, this study reveals the modeling of a newly synthesized polymeric 2D material, and tuning its properties results in smaller bandgaps and half-metallic and magnetic behaviours.

Effect of prostaglandin treatment on the estrus behaviour, follicular and luteal morphometry and serum hormone profile in sub-estrus buffaloes during non-breeding season.

Sub-estrus buffaloes do not exhibit estrus signs despite being cyclic contributing to extended service periods and inter-calving intervals causing significant economic loss. The present study described the effect of synthetic prostaglandin (PGF2α) on estrus behaviour, follicular and luteal morphometry, and serum estradiol (E2) and progesterone (P4) profile in sub-estrus buffaloes during the non-breeding season. The incidence of sub-estrus was 38.4% during the non-breeding season. The sub-estrus buffaloes (n = 33) were divided into two groups, viz., Control (n = 16) and PGF2α treatment (Inj. Cloprostenol 500 µg, i.m., n = 17). Estrus induction response was significantly greater in the treatment (100 vs. 18.75%, p < .001), and a relatively greater proportion of animals conceived in the treatment group (29.41 vs. 6.25%, p = .08). The time elapsed to induction of estrus and insemination following treatment was significantly lower in the treatment group than control. A significant increment in the follicle diameter (9.72 ± 0.45 vs. 13.00 ± 0.45 mm, P < .0001) and serum estradiol (E2) concentration (66.01 ± 11.92 vs. 104.9 ± 13.21 pg/mL, p = .003) observed at the post-treatment period in the PGF2α treatment group. At the same time, CL diameter was reduced significantly at a higher regression rate in the PGF2α treated buffaloes than those of control. Of the responded buffaloes, only 30% showed high-intensity estrus attributed to the expulsion of cervico-vaginal mucus (CVM), uterine tonicity, micturition, and mounting response by a teaser bull. From this study, it can be concluded that the administration of PGF2α could induce estrus in the sub-estrus buffaloes during the non-breeding season. Behavioural changes, along with sonographic observation of POF, regressing CL, and serum E2 and P4 concentration would be useful to determine the right time of insemination in sub-estrus buffaloes during non-breeding season.

Stabilization of NH- Group Adjacent to Naked Silicon(II) Atom in Base Stabilized Aminosilylenes.

Aminosilylene, comprising reactive NH- and Si(II) sites next to each other, is an intriguing class of compounds due to its ability to show diverse reactivity. However, stabilizing the reactive NH- group next to the free Si(II) atom is challenging and has not yet been achieved. Herein, we report the first examples of base stabilized free aminosilylenes Ar*NHSi(PhC(Nt Bu)2 ) (1 a) and Mes*NHSi(PhC(Nt Bu)2 ) (1 b) (Ar*=2,6-dibenzhydryl-4-methylphenyl and Mes*=2,4,6-tri-tert-butylphenyl), tolerating a NH- group next to the naked Si(II) atom. Remarkably, 1 a and 1 b exhibited interesting differences in their reactivity upon heating. With 1 a, an intramolecular C(sp3 )-H activation of one of the benzhydryl methine hydrogen atoms to the Si(II) atom produced the five-membered cyclic silazane 2. However, with 1 b, a rare 1,2-hydrogen shift to the Si(II) atom afforded a silanimine 3, with a hydride ligand attached to an unsaturated silicon atom. Further, the coordination capabilities of 1 a were also tested with Ru(II) and Fe(0) precursors. Treatments of 1 a with [Ru(η6 -p-cymene)Cl2 ]2 led to the isolation of a η6 -arene tethered complex [RuCl2 {Ar*NHSi(PhC(t BuN)2 )-κ1 -Si-η6 -arene}] (4), whereas with the Fe(CO)5 precursor a Fe(0) complex [Fe(CO)4 {Ar*NHSi(PhC(t BuN)2 )-κ1 -Si}] (5) was obtained. Density functional theory (DFT) calculations were conducted to shed light on the structural, bonding, and energetic aspects in 1-5.

2D BN-biphenylene: structure stability and properties tenability from a DFT perspective.

With state-of-the-art density functional theory and ab initio molecular dynamics, we explored the BN-analog of a recently synthesized 2D biphenylene sheet. Its dynamical, thermal, and mechanical stability has been confirmed with phonon spectrum calculations, ab initio molecular dynamics (AIMD), and Born criteria, and its synthesis feasibility has been ascertained in terms of cohesive energy. The phonon spectrum and AIMD results show all positive frequencies and negligible variations in structure (bond length), kinetic and total energy for 5 ps, respectively. Having found this stable BN-BPh, we have analyzed its electronic, mechanical, optical, and vibration properties. Electronic property results (density of states and band structure) show that the BN-equivalent of metallic BPh has a large band gap of 3.12 eV and 4.48 eV with the PBE and HSE06 DFT functionals, respectively, similar to the graphene BN-equivalent (hBN) with a band gap of 6.1 eV. The mechanical strength of BN-BPh is close to that of BPh with a difference of ∼15 N m-1 and ∼10 N m-1 along the x- and y-directions, and its Young's modulus indicates that BN-BPh is stiffer than BN-graphyne. The optical absorption coefficient of BN-BPh shows a first peak at 4.48 eV (3.12 eV) with the HSE06 (PBE) functional. Infrared and Raman spectrum intensity and irreducible representation are computed, and the first IR (Raman) peak is found at 1300 cm-1 (1000 cm-1) for BN-BPh. Finally, substitutions of external atoms such as P, Al, As, C, Li, Mg, S, and Si for B and N atoms in BN-BPh tune its electronic properties to semiconducting, semi-metallic, and even metallic. Considering its mechanical, optical, and band gap tunability, BN-BPh might be useful in electronic, optical, and spintronics devices, even in high-pressure environments.

Functional analysis of metalloenzymes from human gut microbiota and their role in ulcerative colitis.

AIM: Metalloenzymes produced by gut microbiota play an essential role in various physiological processes, and maintains homeostasis of gastrointestinal tract. Our study includes functional analysis of microbial metalloenzymes using metagenomics and metatranscriptomics data from Inflammatory Bowel Disease Multiomics Database. METHODS AND RESULTS: The distance matrix calculated by using metalloenzymes data produced significant results for bacterial taxonomy, with higher variance compared to HMP analysis in both Western and Indian population. Differential gene expression analysis revealed altered expression of ulcerative colitis (UC)-associated enzymes, increased folds changes in Prevotella and Megamonas transcripts; whereas, low transcripts of Alistipes genera. Further, docking and simulation studies performed on screened UC-associated enzymes revealed changes in catalytic efficiency and ligand interacting residues. CONCLUSION: The ß-diversity using microbes containing metalloenzymes suggests considering small group of specific genes or enzymes for understanding the diversity between UC and healthy individuals. The docking and differential gene expression analysis collectively indicate the probable role of metalloenzymes and few UC-associated enzymes in the severity of UC.

Impact of body condition on sow and litter performance, postpartum physiological, hematological, and biochemical parameters in Landlly crossbred pigs.

A total of 32 Landlly crossbred sows were categorized into three groups based on their body condition score (BCS) on the 90th day of gestation viz. low, moderate, and high body condition groups. BCS assessments were subsequently conducted on the 102nd day, at farrowing, and on the 7th, 17th, 27th, and 42nd day postpartum, along with measurements of ultrasonic backfat thickness and body weight. Key reproductive performance traits, including total number born (TNB) and alive (TLA), stillbirths, mummification, and litter weight at birth and weaning, were recorded. Several physiological, health, biochemical, and hematological parameters were also estimated at different lactation stages. Analysis of the data revealed that TNB, TLA (P<0.04), and weaned piglets (P<0.01), as well as litter weight at birth (P<0.08) and weaning (P<0.08), were significantly higher in the low BCS group. Meanwhile, the weaning to estrous interval (P<0.04) was optimized in the high and moderate BCS groups. However, sows with high body condition exhibited higher stillbirths and pre-weaning mortality. Additionally, the order of increasing body condition correlated with superior colostrum quality, characterized by higher IgG levels (P<0.02), and increased average milk yield during early and mid-lactation (P<0.03). A high occurrence of postpartum dysgalactia syndrome (PDS) was evident in obese sows (P<0.001). Moreover, a positive correlation was found between the body condition of sows at late gestation and cortisol concentration throughout lactation across the three BCS groups (P<0.0001). Furthermore, a strong positive correlation was observed between the postpartum serum concentration of T3 (thyroid hormone) (P<0.002) and the prepartum body condition of sows. Based on these findings, maintaining sows with a moderate body condition (BCS of 3) and a backfat thickness of 21 mm in the breeding herd is recommended for enhancing profitability and productivity at the farm level.

Efficient Recycling of Gold and Copper from Electronic Waste by Selective Precipitation.

The recycling of metals from electronic waste (e-waste) using efficient, selective, and sustainable processes is integral to circular economy and net-zero aspirations. Herein, we report a new method for the selective precipitation of metals such as gold and copper that offsets the use of organic solvents that are traditionally employed in solvent extraction processes. We show that gold can be selectively precipitated from a mixture of metals in hydrochloric acid solution using triphenylphosphine oxide (TPPO), as the complex [(TPPO)4 (H5 O2 )][AuCl4 ]. By tuning the acid concentration, controlled precipitation of gold, zinc and iron can be achieved. We also show that copper can be selectively precipitated using 2,3-pyrazinedicarboxylic acid (2,3-PDCA), as the complex [Cu(2,3-PDCA-H)2 ]n ⋅ 2n(H2 O). The combination of these two precipitation methods resulted in the recovery of 99.5 % of the Au and 98.5 % of the Cu present in the connector pins of an end-of-life computer processing unit. The selectivity of these precipitation processes, combined with their straightforward operation and the ability to recycle and reuse the precipitants, suggests potential industrial uses in the purification of gold and copper from e-waste.

Faecal microbiota transplantation with anti-inflammatory diet (FMT-AID) followed by anti-inflammatory diet alone is effective in inducing and maintaining remission over 1 year in mild to moderate ulcerative colitis: a randomised controlled trial.

OBJECTIVE: Microbiome and dietary manipulation therapies are being explored for treating ulcerative colitis (UC). We aimed to examine the efficacy of multidonor faecal microbiota transplantation (FMT) and anti-inflammatory diet in inducing remission followed by long-term maintenance with anti-inflammatory diet in patients with mild-moderate UC. DESIGN: This open-labelled randomised controlled trial (RCT) randomised patients with mild-moderate (Simple Clinical Colitis Activity Index (SCCAI) 3-9) endoscopically active UC (Ulcerative Colitis Endoscopic Index of Severity (UCEIS)>1) on stable baseline medications in 1:1 ratio to FMT and anti-inflammatory diet (FMT-AID) versus optimised standard medical therapy (SMT). The FMT-AID arm received seven weekly colonoscopic infusions of freshly prepared FMT from multiple rural donors(weeks 0-6) with anti-inflammatory diet. Baseline medications were optimised in the SMT arm. Clinical responders (decline in SCCAI>3) at 8 weeks in both arms were followed until 48 weeks on baseline medications (with anti-inflammatory diet in the FMT-AID arm). Primary outcome measures were clinical response and deep remission (clinical-SCCAI <2; and endoscopic-UCEIS <1) at 8 weeks, and deep remission and steroid-free clinical remission at 48 weeks. RESULTS: Of the 113 patients screened, 73 were randomised, and 66 were included in (35-FMT-AID; 31-SMT) modified intention-to-treat analysis (age-35.7±11.1 years; male-60.1%; disease duration-48 (IQR 24-84) months; pancolitis-34.8%; SCCAI-6 (IQR 5-7); UCEIS-4 (IQR 3-5)). Baseline characteristics were comparable. FMT-AID was superior to SMT in inducing clinical response (23/35 (65.7%) vs 11/31 (35.5%), p=0.01, OR 3.5 (95% CI 1.3 to 9.6)), remission (21/35 (60%) vs 10/31 (32.3%), p=0.02, OR 3.2 (95% CI 1.1 to 8.7)) and deep remission (12/33 (36.4%) vs 2/23 (8.7%), p=0.03, OR 6.0 (95% CI 1.2 to 30.2)) at 8 weeks. Anti-inflammatory diet was superior to SMT in maintaining deep remission until 48 weeks (6/24 (25%) vs 0/27, p=0.007). CONCLUSION: Multidonor FMT with anti-inflammatory diet effectively induced deep remission in mild-moderate UC which was sustained with anti-inflammatory diet over 1 year. TRIAL REGISTRATION NUMBER: ISRCTN15475780.

AnAb-initiostudy of the Y decorated 2D holey graphyne for hydrogen storage application.

Expanding pollution and rapid consumption of natural reservoirs (gas, oil, and coal) led humankind to explore alternative energy fuels like hydrogen fuel. Solid-state hydrogen storage is most desirable because of its usefulness in the onboard vehicle. In this work, we explored the yttrium decorated ultra porous, two-dimensional holey-graphyne for hydrogen storage. Using the first principles density functional theory simulations, we predict that yttrium doped holey graphyne can adsorb up to seven hydrogen molecules per yttrium atom resulting in a gravimetric hydrogen weight percentage of 9.34, higher than the target of 6.5 wt% set by the US Department of Energy. The average binding energy per H2and desorption temperature come out to be -0.34 eV and ∼438 K, respectively. Yttrium atom is bonded strongly on HGY sheet due to charge transfer from Y 4d orbital to C 2p orbital whereas the adsorption of H2molecule on Y is due to Kubas-type of interactions involving charge donation from H 1s orbital to Y 3d orbital and back donation with net charge gain by H 1s orbital. Furthermore, sufficient energy barriers for the metal atom diffusion have been found to prevent the clustering of transition metal (yttrium) on HGY sheet. The stability of the system at higher temperatures is analyzed usingAb-initiomolecular dynamics (AIMD) method, and the system is found to be stable at room and the highest desorption temperature. Stability of the system at higher temperatures, presence of adequate diffusion energy barrier to prevent metal-metal clustering, high gravimetric wt% of H2uptake with suitable binding energy, and desorption temperature signifies that Y doped HGY is a promising material to fabricate high capacity hydrogen storage devices.

Effect of Vitamin D Supplementation on CD4 Count in HIV-Infected Children and Adolescents in North India: A Non-Randomized Comparative Study.

BACKGROUND: HIV infection is still a serious public health issue globally. Suboptimal vitamin D status is highly prevalent in HIV-infected children and adolescents throughout the world. OBJECTIVES: To evaluate the outcome of vitamin D supplementation on CD4 count in HIV-infected children and adolescents with suboptimal vitamin D status. METHODS: Vitamin D level of HIV-infected children and adolescents were measured at enrolment. Suboptimal vitamin D level was defined as 25(OH)D < 30 ng/ml. Vitamin D insufficiency and deficiency were defined as 21-29 and <20 ng/ml, respectively. Children with suboptimal vitamin D levels were supplemented with vitamin D. RESULTS: This was a single-centre, non-randomized comparative study enrolling 50 eligible participants. There were 20 patients who were vitamin D sufficient, 7 were vitamin D insufficient and 23 were found to be vitamin D deficient at enrolment. However, after supplementation, the status of sufficient remained same and 7 insufficient become sufficient, whereas in 23 deficient, 18 (78.3%) become sufficient and 5 (21.7%) become insufficient and this change was found statistically significant among the groups (χ2 = 6.52, p = 0.038). There was a significant improvement of CD4 count from baseline to 4 months in deficient group on vitamin D supplementation (p value < 0.001; 1.2-fold rise). No significant change was seen in vitamin D insufficient (p value = 0.791) and sufficient groups (p value = 0.168). CONCLUSION: Vitamin D should be supplemented in HIV-infected children on ART with low CD4 counts.

Nebulized Salbutamol with or without Magnesium Sulphate in the Management of Acute Asthma in Children in India: A Randomized Controlled Trial.

BACKGROUND: There is inconclusive evidence on the role of nebulized magnesium sulphate (MgSO4) in the management of acute asthma in paediatric population. OBJECTIVES: Whether the use of nebulized salbutamol with or without MgSO4 in the management of acute asthma results in clinically significant improvement in lung function in Indian children? The primary outcome measure was to assess improvements in peak expiratory flow rate (PEFR), heart rate, respiratory rate and SpO2. METHODS: This was a single centre; prospective double-blind randomized control trial conducted in paediatric intensive care unit of a tertiary care centre. Ninety children of 6-14 years with acute exacerbations of bronchial asthma were enrolled to receive either inhaled magnesium sulphate (95 mg) with salbutamol (5 mg) or inhaled salbutamol (5 mg) alone. All patients got three nebulizations done during the first hour at 20 min intervals, two nebulizations during the second hour at 30 min intervals, hourly for the next 2 h and then at 24 and 48 h. RESULTS: Eighty-five patients were finally analysed as per protocol analysis. The trial showed that PEFR increased gradually in both groups over the study duration, but it was statistically not significant. Heart rate decreased significantly in both groups over the study duration. Respiratory rate decreased significantly between the groups at 24 and 48 h only. SpO2 improved too in both groups but was not significant statistically. CONCLUSION: The addition of nebulized MgSO4 to salbutamol does not seem to result in improvement in lung function in the management of acute asthma in Indian children.

Effect of milking environment enrichment through music on production performance and behaviour in cattle.

Enrichment of milking environment through music has been proposed to help animals to cope with divergent stressors. In sight of the above, a study was conducted to evaluate the effect of Indian instrumental music-based environmental enrichment played in yaman raga on milk production performance and behaviour in cattle. A total of 21 lactating dairy cattle (Vrindavani crossbred cows) having similar parity and stage of lactation were selected in three groups - T1, T2 and T3, each consisting of seven animals. The T1 and T2 groups were exposed to instrumental flute and sitar, respectively, 10 min prior to the start of milking and continued till completion of milking; while the T3 group served as control. Musical enrichment of the environment was done using recorded-tape of flute and sitar was played in yamen raga at 40-60 (dB) decibel intensity. The results revealed a non-significant difference in milk yield, rectal temperature, respiration rate, T3 (triiodothyronine) and T4 (thyroxine) hormones. However, there exhibited a significant (p < 0.05) difference in milking time, milking speed, cortisol hormones and behavioural parameters such as milk let-down in the animals exposed to music compared to the control group. Thus, the results have significant implications relating to the behavioural fitness and welfare of dairy animals and reducing residual milk.

A SUMOylation-dependent switch of RAB7 governs intracellular life and pathogenesis of Salmonella Typhimurium.

Salmonella Typhimurium is an intracellular pathogen that causes gastroenteritis in humans. Aided by a battery of effector proteins, S. Typhimurium resides intracellularly in a specialized vesicle, called the Salmonella-containing vacuole (SCV) that utilizes the host endocytic vesicular transport pathway (VTP). Here, we probed the possible role of SUMOylation, a post-translation modification pathway, in SCV biology. Proteome analysis by complex mass-spectrometry (MS/MS) revealed a dramatically altered SUMO-proteome (SUMOylome) in S. Typhimurium-infected cells. RAB7, a component of VTP, was key among several crucial proteins identified in our study. Detailed MS/MS assays, in vitro SUMOylation assays and structural docking analysis revealed SUMOylation of RAB7 (RAB7A) specifically at lysine 175. A SUMOylation-deficient RAB7 mutant (RAB7K175R) displayed longer half-life, was beneficial to SCV dynamics and functionally deficient. Collectively, the data revealed that RAB7 SUMOylation blockade by S. Typhimurium ensures availability of long-lived but functionally compromised RAB7, which was beneficial to the pathogen. Overall, this SUMOylation-dependent switch of RAB7 controlled by S. Typhimurium is an unexpected mode of VTP pathway regulation, and unveils a mechanism of broad interest well beyond Salmonella-host crosstalk. This article has an associated First Person interview with the first author of the paper.

A Dicobalt(II) Single-Molecule Magnet via a Well-Designed Dual-Capping Tetrazine Radical Ligand.

The recent years have witnessed the glory development for the construction of high-performance mononuclear single molecule magnets (SMMs) within a specific coordination geometry, which, however, is not well applied in cluster-based SMMs due to the synthetic challenges. Given that the monocobalt(II) complexes within a trigonal-prismatic (TPR) coordination geometry have been classified as excellent SMMs with huge axial anisotropy (D ≈ -100 cm-1), here we designed and synthesized a new dual-capping tetrazine ligand, 3,6-bis(6-(di(1H-pyrazol-1-yl)methyl)pyridin-2-yl)-1,2,4,5-tetrazine (bpptz), and prepared a novel dicobalt(II) complex, [Cp2CoIII][{(hfac)CoII}2(bpptz•-)][hfac]2·2Et2O (1, hfac = hexafluoroacetylacetonate). In the structure of 1, the bpptz•- radical ligand enwraps two Co(II) centers within quasi-TPR geometries, which are further bridged by the tetrazine radical in the trans mode. The magnetic study revealed that the interaction between the Co centers and the tetrazine radical is strongly antiferromagnetic with a coupling constant (J) of -65.8 cm-1 (in the -2J formalism). Remarkably, 1 exhibited the typical SMM behavior with an effective energy barrier of 69 cm-1 under a 1.5 kOe dc field, among the largest for polynuclear transition metal SMMs. In addition, DFT and ab initio calculations suggested that the presence of a strong Co(II)-radical magnetic interaction effectively quenches the QTM effect and enhances the barrier height for the magnetization reversal.

Clinical outcome of living donor kidney transplantation across simultaneous ABO and HLA incompatibility: Single center experience of first ten cases.

BACKGROUND AND AIMS: Preconditioning using different protocols has been tested to prevent antibody mediated rejection (ABMR) individually for ABO and HLA incompatibility. However, simultaneous presence of both barriers is still less explored. The aim of this study was to report outcomes of institutional desensitization protocol in renal transplant recipients with simultaneous ABO and HLA incompatibility. MATERIALS AND METHODS: This was a retrospective study conducted from October 2015 to December 2018. All patients with a clinical diagnosis of dialysis dependent chronic kidney disease (CKD), who were prospective coexistent HLA and ABO incompatible renal transplant recipients were included in the study. Patients were followed up and graft function and patient survival was assessed at 1 y from the date of transplant. RESULTS: Median and mode baseline anti-A titers were 64, while median and mode baseline anti-B titers were 256. All recipients were discharged by tenth postoperative day. None of the patients had any bleeding complications. Post transplant infection rate was found to be 20 %. A total of 54 therapeutic plasma exchange (TPE) procedures were performed before transplant and 8 were performed after transplant. Graft survival and patient survival was 100 % at 3, 6, 9, and 12 months. Range and mean follow-up period was 15-42 months and 23 months respectively. Mean glomerular filtration rate (GFR) at 1 y using the CKD-EPI equation was 85.25 ± 13.76 mL/min. Biopsy proven ABMR was observed in one case only which was managed with TPE and immunosuppression. CONCLUSION: Simultaneous ABO and HLA incompatibility in renal transplant recipients can be managed successfully with adequate preconditioning and careful monitoring.

Prospective audit of blood transfusion request forms and continuing medical education to optimise compliance of clinicians in a hospital setting.

OBJECTIVES: The aim of this study was to analyse blood requisition forms sent by clinicians in a tertiary care hospital to the transfusion service to ascertain their completeness and correctness. A secondary objective was to study the effect of continuing medical education (CME) in a hospital setting on clinician's behaviour regarding the importance of details that ought to be mentioned on blood requisition forms. BACKGROUND: Transfusion audits are useful tools in the evaluation and education of those requesting blood components. METHODS/MATERIALS: This was a prospective, observational study conducted in the department of Transfusion Medicine at a tertiary-level healthcare centre from June 2019 to December 2019. The study was divided into two phases: pre-CME (P1) and post-CME (P2). In both phases, an audit for assessing completeness and correctness of blood requisition forms, which were divided into four sections, was performed. A scoring system was devised to compare both phases. RESULTS: In the P1 phase, 45.77% of the blood requisition form entries were complete and correct; 23.45% of incomplete entries were generated by emergency and trauma. In the P2 phase, 76.75% of the blood requisition form entries were complete and correct; 35.09% of the incomplete entries were generated by obstetrics and gynaecology. Complete and correct entries increased from 45.7% (P1) to 76.75% (P2). Scores of P1 were found to be lower than scores of P2 for all four sections. Cumulative mean score for P1 (20687) was found to be significantly lower than the mean score for P2 (30870). CONCLUSION: Audit and CME regarding different aspects of transfusion medicine practices play a major role in the improvement of transfusion practices in hospitals.

Outcome of desensitization in human leukocyte antigen and ABO incompatible living donor kidney transplantation: Single center experience of first 200 incompatible transplants.

BACKGROUND AND AIMS: Although desensitization is well established, concerns about graft outcome, patient survival and rejection still exist. The present study aims at comparing outcomes of renal transplant recipients across simultaneous ABO and human leukocyte antigen (HLA) incompatibility barriers to those with ABO or HLA incompatibility alone. MATERIALS AND METHODS: This was a retrospective study conducted from October 2015 to December 2018. All patients with a clinical diagnosis of chronic kidney disease, who were prospective HLA incompatible (HLAi) and/or ABO incompatible (ABOi) renal transplant recipients were included. A total of 400 cases including 36 ABOi transplants, 154 HLAi transplants, 10 simultaneously ABO and HLA incompatible transplants, and 200 ABO (ABOc) and HLA (HLAc) compatible kidney transplants from living donors were included. RESULTS: There were significantly more number of blood transfusions, previous transplants and pregnancies in HLAi transplant recipients relative to the ABOi or the control group. Mean number of therapeutic plasma exchange procedures per patient and mean plasma volume processed per procedure were slightly higher in the ABOi + HLAi category. The incidence of graft dysfunction due to suspected antibody-mediated rejection during first year was highest in the ABOi + HLAi group, followed by ABOc + HLAi and ABOi + HLAc, lowest in the ABOc + HLAc category. Mean time to first episode of graft dysfunction was significantly shorter with incompatible transplants. There were no kidney transplant recipient deaths in the study. CONCLUSION: Patient outcome and graft outcomes observed with incompatible transplants were not worse than those observed with compatible transplants.