Noncanonical binding of Lck to CD3ε promotes TCR signaling and CAR function.

Initiation of T cell antigen receptor (TCR) signaling involves phosphorylation of CD3 cytoplasmic tails by the tyrosine kinase Lck. How Lck is recruited to the TCR to initiate signaling is not well known. We report a previously unknown binding motif in the CD3ε cytoplasmic tail that interacts in a noncanonical mode with the Lck SH3 domain: the receptor kinase (RK) motif. The RK motif is accessible only upon TCR ligation, demonstrating how ligand binding leads to Lck recruitment. Binding of the Lck SH3 domain to the exposed RK motif resulted in local augmentation of Lck activity, CD3 phosphorylation, T cell activation and thymocyte development. Introducing the RK motif into a well-characterized 41BB-based chimeric antigen receptor enhanced its antitumor function in vitro and in vivo. Our findings underscore how a better understanding of the functioning of the TCR might promote rational improvement of chimeric antigen receptor design for the treatment of cancer.

Constitutive expression and discovery of antimicrobial peptides in Zygogramma bicolorata (Coleoptera: Chrysomelidae).

The expression, identification, and discovery of less toxic antimicrobial peptides (AMPs) are significant in managing infectious pathogens. AMPs triggered in response to the immune system have evolved to defend against pathogens and wounding. The protein composition of Zygogramma bicolorata hemolymph is of diagnostic importance as the open circulatory systems of the insects involve signaling through hemolymph. They have conserved many ancestral vertebrate genes that may help better understand the evolution of innate immunity. The present work describes the isolation, purification, identification, and bioinformatics analysis of AMPs from the immunized hemolymph of Z. bicolorata. Thirty-nine peptides were isolated from reverse-phase high-performance liquid chromatography and sequenced via mass spectrometry analysis. The immunization process recorded a threefold higher protein concentration in immunized hemolymph when compared with nonimmunized one. For the first time, the proteomic study on Z. bicolorata hemolymph unveils the three novel proteins in the family Chrysomelidae with no homology in the database, indicating its novelty and the expression of the rest of 36 well-known proteins, including heat-shock, immune, structural, signaling proteins, and others speak for its method validity. Combining the expression of novel AMPs, detoxifying enzymes, hemolytic, and cytotoxic assays, and this work can elucidate new pathways to immune response mechanisms. Its molecular basis also holds the potential applicability in the future drug development process against pathogenic fungi such as Aspergillus niger and Candida albicans.

Dichotomy between the humoral and cellular responses elicited by mRNA and adenoviral vector vaccines against SARS-CoV-2.

BACKGROUND: Protection from severe disease and hospitalization by SARS-CoV-2 vaccination has been amply demonstrated by real-world data. However, the rapidly evolving pandemic raises new concerns. One pertains efficacy of adenoviral vector-based vaccines, particularly the single-dose Ad26.COV2.S, relative to mRNA vaccines. MAIN BODY: We investigated the immunogenicity of Ad26.COV2.S and mRNA vaccines in 33 subjects vaccinated with either vaccine class 5 months earlier on average. After controlling for the time since vaccination, Spike-binding antibody and neutralizing antibody levels were higher in the mRNA-vaccinated subjects, while no significant differences in antigen-specific B cell and T cell responses were observed between the two groups. CONCLUSIONS: A dichotomy exists between the humoral and cellular responses elicited by the two vaccine classes. Testing only for humoral responses to compare the durability of SARS-CoV-2 vaccine-induced responses, as typically performed for public health and research purposes, is insufficient.

Native nephrectomy in the management of hypertension in children with kidney disease: a tool to improve blood pressure control.

AIM: Hypertension in children with abnormal kidneys often requires multiple antihypertensive agents (complex), or could present with complications (e.g. hypertensive encephalopathy). Our objective in this report is to evaluate blood pressure control following unilateral or bilateral laparoscopic native nephrectomy in children with renal hypertension. MATERIALS AND METHODS: Single-centre retrospective review of all children who underwent nephrectomy for management of hypertension over a recent study period (2008-2017) with post-operative follow-up of at least 3 years. We describe the association of age, primary kidney disease and blood pressure and its management including time to resolution following unilateral or bilateral nephrectomy. RESULTS: During the 9-year study period, 21 of 215 (9.8%) children underwent nephrectomy for management of hypertension. We included 19 children [6 with unilateral native nephrectomy (UNN) and 13 with bilateral native nephrectomy (BNN)] in this study as they continued with their follow-up at our centre. Out of the 19 children, 15 had laparoscopic retroperitoneoscopic nephrectomies and 4 had laparoscopic transperitoneal nephrectomies. Six children had unilateral nephrectomy and 13 children had bilateral nephrectomies [7 were pre-transplant (haemodialysis-6, peritoneal dialysis-1) and 6 were post-kidney transplant]. Fifteen of 19 children (79%) had complete resolution [5 UNN and 10 BNN] and 3 (16%) partial resolution [1 UNN and 2 BNN]. One patient with BNN was observed to have no change in blood pressure control. CONCLUSION: Our data demonstrate improved management of hypertension in 95% of the children. Nephrectomy could offer a reasonable treatment option for selected group of complex and complicated renal hypertension.

Sterile particle-induced inflammation is mediated by macrophages releasing IL-33 through a Bruton's tyrosine kinase-dependent pathway.

Initiation of the innate sterile inflammatory response that can develop in response to microparticle exposure is little understood. Here, we report that a potent type 2 immune response associated with the accumulation of neutrophils, eosinophils and alternatively activated (M2) macrophages was observed in response to sterile microparticles similar in size to wear debris associated with prosthetic implants. Although elevations in interleukin-33 (IL-33) and type 2 cytokines occurred independently of caspase-1 inflammasome signalling, the response was dependent on Bruton's tyrosine kinase (BTK). IL-33 was produced by macrophages and BTK-dependent expression of IL-33 by macrophages was sufficient to initiate the type 2 response. Analysis of inflammation in patient periprosthetic tissue also revealed type 2 responses under aseptic conditions in patients undergoing revision surgery. These findings indicate that microparticle-induced sterile inflammation is initiated by macrophages activated to produce IL-33. They further suggest that both BTK and IL-33 may provide therapeutic targets for wear debris-induced periprosthetic inflammation.

The potential of arbuscular mycorrhizal fungi in C cycling: a review.

Arbuscular mycorrhizal fungi (AMF) contribute predominantly to soil organic matter by creating a sink demand for plant C and distributing to below-ground hyphal biomass. The extra-radical hyphae along with glomalin-related soil protein significantly influence the soil carbon dynamics through their larger extent and turnover period need to discuss. The role of AMF is largely overlooked in terrestrial C cycling and climate change models despite their greater involvement in net primary productivity augmentation and further accumulation of this additional photosynthetic fixed C in the soil. However, this buffering mechanism against elevated CO2 condition to sequester extra C by AMF can be described only after considering their potential interaction with other microbes and associated mineral nutrients such as nitrogen cycling. In this article, we try to review the potential of AMF in C sequestration paving the way towards a better understanding of possible AMF mechanism by which C balance between biosphere and atmosphere can be moved forward in more positive direction.

Arbuscular mycorrhiza: a viable strategy for soil nutrient loss reduction.

Arbuscular mycorrhiza fungi's (AMF) role in plant nutrition and stress management is well known, but very few researches and studies have been conducted so far on the fungal ability to reduce different nutrient losses (runoff, leaching and volatilization) from the soil system. This important ecosystem service of AMF had been neglected largely. From the recent findings, it has been confirmed that mycorrhizal symbiosis has potential to check the losses of applied nutrients. The role of soil biota in nutrient cycling is indispensable and determines the nutrient availability to plants. Among these biota, AMF's association with plants is the most prevalent, but the exact mechanisms followed by AMF in nutrient cycling, transformation and reducing nutrient loss ability are still inconclusive. In this review, we will try to unlock this particular aspect of AMF which is important to achieve global food demand in a sustainable way.

Anti-cholinesterase hybrids as multi-target-directed ligands against Alzheimer's disease (1998-2018).

Alzheimer's disease (AD) is a genetically complex, progressive and irreversible neurodegenerative disorder of the brain which involves multiple associated etiological targets. The complex pathogenesis of AD gave rise to multi-target-directed ligands (MTDLs) principle to combat this dreaded disease. Within this approach, the design and synthesis of hybrids prevailed greatly because of their capability to simultaneously target the intertwined pathogenesis components of the disease. The hybrids include pharmacophoric hybridization of two or more established chemical scaffolds endowed with the desired pharmacological properties into a single moiety. In AD, the primary foundation of medication therapy and drug design strategies includes the inhibition of cholinesterase (ChE) enzymes. Hence the development of ChE inhibition based hybrids is the central choice of AD medicinal chemistry research. To illustrate the progress of ChE inhibition based hybrids and novel targets, we reviewed the medicinal chemistry and pharmacological properties of the multi-target molecules published since 1998-December 2018. We hope that this article will allow the readers to easily follow the evolution of this prominent medicinal chemistry approach to develop a more efficient inhibitor.

Bromodomain Drug Discovery - the Past, the Present, and the Future.

With the bromodomain (BRD) inhibitor JQ1, a remarkable success story of BRD4 as a novel drug target has been set off that yielded many anti-cancer drugs that are now in clinical trials. But not all of the great prospects of BRDs as drug targets may become true. First evaluations of ongoing clinical trials revealed that treatment with BET-inhibitors can be accompanied with significant toxic side effects and the validation of the therapeutic benefit of BET-inhibitors compared to existing therapies is still pending. New strategies that may overcome possible obstacles in BRD drug discovery include combination therapies with other agents, dual target inhibitors, and proteolysis targeting chimeras (PROTACs). Furthermore, non-BET proteins seem promising drug targets as well. Most recently, BRDs have been identified as putative targets to treat parasitic diseases such as malaria. Milestones in BRD drug discovery are reviewed and promising new developments are evaluated.

Prospects of Using High-Intensity THz Pulses To Induce Ultrafast Temperature-Jumps in Liquid Water.

Ultrashort, high-intensity terahertz (THz) pulses, e.g., generated at free-electron laser facilities, allow for direct investigation as well as the driving of intermolecular modes in liquids like water and thus will deepen our understanding of the hydrogen bonding network. In this work, the temperature-jump (T-jump) of water induced by THz radiation is simulated for ten different THz frequencies in the range from 3 to 30 THz and five different pulse intensities in the range from 1 × 1011 to 5 × 1012 W/cm2 employing both ab initio molecular dynamics (AIMD) and force field molecular dynamics (FFMD) approaches. The most efficient T-jump can be achieved with 16 THz pulses. Three distinct T-jump mechanisms can be uncovered. For all cases, the T-jump mechanism proceeds within tens of femtoseconds (fs). For frequencies between 10 and 25 THz, most of the energy is initially transferred to the rotational degrees of freedom. Subsequently, the energy is redistributed to the translational and intramolecular vibrational degrees of freedom within a maximum of 500 fs. For the lowest frequencies considered (7 THz and below), translational and rotational degrees of freedom are heated within tens of fs as the THz pulse also couples to the intermolecular vibrations. Subsequently, the intramolecular vibrational modes are heated within a few hundred fs. At the highest frequencies considered (25 THz and above), vibrational and rotational degrees of freedom are heated within tens of fs, and energy redistribution to the translational degrees of freedom happens within several hundred fs. Both AIMD and FFMD simulations show a similar dependence of the T-jump on the frequency employed. However, the FFMD simulations overestimate the total energy transfer around the main peak and drop off too fast toward frequencies higher and lower than the main peak. These differences can be rationalized by missing elements, such as the polarizability, in the TIP4P/2005f force field employed. The feasibility of performing experiments at the studied frequencies and intensities as well as important issues such as energy efficiency, penetration depth, and focusing are discussed.

Freezing transitions in a system of two-dimensional octupolar multipoles.

We have investigated the fluid-solid freezing transitions in a system of axially symmetric particles confined to a two-dimensional plane and interacting via purely repulsive octupolar interaction potential varying as the seventh power of the inverse interparticle separation. Both the one-component and two-component cases have been considered. The classical density functional theory of freezing has been employed to study the relative stability of the triangular solid phase of the system with respect to the fluid phase of the system using the structural inputs calculated by solving the Rogers-Young integral equation theory. Considering the freezing of the fluid into substitutionally disordered solid, in the case of binary mixtures, we observe that the temperature-composition phase diagram is a spindle for moderate particle asymmetries in the range 0.90-0.75. Further increasing the asymmetry to 0.70 results in the coexistence of the fluid phases of two different compositions.

A modified technique of laparoscopic mobilisation and transfer of intra-abdominal testis via inguinal canal.

Several techniques have been described for mobilising intra-abdominal testis (IAT) into the scrotum. Many are associated with significant morbidity. We hereby introduce our modification of the technique that is safe and reproducible. Surgical technique using a nylon tape to railroad instruments into the peritoneal cavity to minimise false passage and trauma is described in detail. Once the testis is delivered out of the scrotal wound, further division of restrictive bands to mobilise the testis is carried out laparoscopically. These steps are not previously described in the literature. A retrospective study of consecutive patients with IAT undergoing surgical intervention by a single surgeon over a period of 15 years has been included to demonstrate outcome. Twenty-five patients with unilateral IAT (10R, 15L) and 2 patients with bilateral IAT, a total of 29 IAT, underwent laparoscopic orchidopexy over the period 1997-2012. Median age of the patients was 3.0 years. One-stage testicular vessel preserving (VP) orchidopexy was performed in 20 testes, Fowler-Stephen (FS), vessel sacrificing orchidopexy (3 one-stage and 6 two-stage) was required in 9 testes. At follow-up, atrophy was detected in 1 (5.0 %) and 2 (22.2 %) patients of the VP and FS groups respectively. The railroading technique of transferring IAT into the scrotum via the inguinal canal under laparoscopic guidance minimises tissue trauma. It negates the need to create new tissue opening. It is reproducible with a testicular atrophy rate comparable to published literature.

Metabolism of cisplatin in the organs of Rattus norvegicus: role of Glutathione S-transferase P1.

Glutathione S-transferases (GSTs) play an important role in the biotransformation of endogenous compounds and xenobiotics as well as in the metabolic inactivation of pharmacologically active substances, including anticancer drugs. Using cisplatin as the prototype drug, we investigated if any correlation exists between GSH levels, GSTs/GSTP1 activity and the fate of cisplatin in different organs of Rattus norvegicus. GSH-cisplatin complex was prepared, purified by anion-exchange chromatography and subjected to mass spectroscopic analysis which confirmed the structure to be diglutathione-monoplatinum (diglutathionylplatinum). Purified diglutathionylplatinum was used to quantify metabolite formed in different tissue homogenates. Specific GSTP1 activity was found to be highest in kidneys, which correlated positively with the levels of metabolite formed in renal tissues. Altogether, our results showed that cisplatin metabolism in different organs of rats correlated positively with specific GSTP1 activities and this enzyme may be a critical determinant of extent of cellular uptake or retention of cisplatin in renal and liver tissues.

Extraoral plasmablastic lymphoma detected using ascitic fluid cytology and flow cytometry: a case report with a review of the literature.

BACKGROUND: Plasmablastic lymphoma (PL) is a relatively new category of lymphoma, which has been considered to be found predominantly in the oral cavity and has a strong association with HIV. CASE: We report a case of extraoral/mesenteric PL detected using cytological examination of ascitic fluid assisted by flow cytometric (FC) analysis. The cells were positive for CD38, CD138, CD10, CD45 and CD56 and negative for CD3, CD19, CD20 and CD79a, with cytoplasmic lambda light-chain restriction. We also reviewed 67 cases of extraoral PL from the available literature and found them to be less often associated with HIV (than oral PL), occurring mostly in males aged 30-60 years, with the most common extraoral site being the anorectal region. CONCLUSION: A high index of suspicion at the level of the cytopathologist is imperative for identifying lymphoma cells in a body fluid. A rare entity like PL can also be diagnosed on cytology assisted by ancillary techniques (like FC), without the need for a biopsy. We also suggest that the minimum panel to diagnose PLs should include CD138, MUM-1, Ki-67, ALK-1, CD3, immunoglobulin light-chains, CD20 and PAX5.

Incarcerated inguinal hernia management in children: 'a comparison of the open and laparoscopic approach'.

PURPOSE: To compare the outcomes of management of incarcerated inguinal hernia by open versus laparoscopic approach. METHODS: This is a retrospective analysis of incarcerated inguinal hernina in a paediatric surgery centre involving four consultants. Manual reduction was attempted in all and failure was managed by emergency surgery. RESULTS: The laparoscopy group had 27 patients. Four patients failed manual reduction and underwent emergency laparoscopic surgery. Three of them had small bowel strangulation which was reduced laparoscopically. The strangulated bowel was dusky in colour initially but changed to normal colour subsequently under vision. The fourth patient required appendectomy for strangulated appendix. One patient had concomitant repair of umbilical hernia and one patient had laparoscopic pyloromyotomy at the same time. One patient had testicular atrophy, one had hydrocoele and one had recurrence of hernia on the asymptomatic side. The open surgery group had 45 patients. Eleven patients had failed manual reduction requiring emergency surgery, of these two required resection and anastomosis of small intestine. One patient in this group had concomitant repair of undescended testis. There was no recurrence in this group, one had testicular atrophy and seven had metachronous hernia. CONCLUSIONS: Both open herniotomy and laparoscopic repair offer safe surgery with comparable outcomes for incarcerated inguinal hernia in children. Laparoscopic approach and hernioscopy at the time of open approach appear to show the advantage of repairing the contralateral patent processus vaginalis at the same time and avoiding metachronous inguinal hernia.

Laparoscopic strategies in complex upper urinary tract obstruction.

OBJECTIVES: We enumerate the various laparoscopic strategies to resolve upper urinary tract (UUT) obstruction in the context of variations in anatomy and report their outcomes. PATIENTS AND METHODS: Retrospective review of primary laparoscopic UUT reconstructions performed between May 2012 and May 2021. Anomalies included: malrotated kidney (MRK), horseshoe kidney (HSK), duplex kidney (DK), pure intrarenal pelvis (IRP) and mid-ureteric stenosis (MUS). Success was defined by postoperative resolution of symptoms, improvement of anterior-posterior renal pelvic diameter (APD) on US and drainage on Mag3. Complications were categorised by Clavien-Dindo grading. Outcomes compared using the student's t-test with P < 0.05 considered statistically significant. RESULTS: Of the 214 laparoscopic primary UUT reconstructions, 37 (17.2 %) were: 13-MRK, 12-HSK, 4-DK, 4-IRP and 4-MUS. Median age at surgery was 5-years (range 0.3-15). Dismembered pyeloplasty: 8; pyeloplasty with renal sinus dissection: 8; neo-PUJ anastomosis: 8; primary ureterocalycostomy: 7; pyeloureterostomy: 2; and uretero-ureterostomy: 4. Median follow-up was 43-months (range 8-108) with a success rate of 94.5 % (35/37). Complete resolution of symptoms in 20/21 patients; improvement of hydronephrosis on US in 35/37 patients (median pre-operative APD 27 mm vs. median postoperative APD 8 mm) [P < 0.001]; improvement of drainage on diuretic renogram in 32/34 kidneys and stable/improved DRF in 34/35 kidneys (median preoperative DRF - 45 % vs. median postoperative DRF - 47 %) [P > 0.05]. Postoperative complications managed medically (II Clavien) included urinary tract infections - 2 patients (5 %), stent-related symptoms in 2 (5 %) and umbilical port site collection in 1 patient (3 %). Recurrent pelvi-ureteric junction obstruction PUJO occurred in one patient (3 %) requiring redo surgery (IIIb Clavien), renal stones in 1 (3 %) which resolved with ESWL (IIIb Clavien); in 1 (3 %) patient with a HSK there was complete loss of ipsilateral kidney function but this was managed conservatively up to date (I Clavien). CONCLUSION: Laparoscopic transperitoneal approach allows the prompt recognition of in-situ anatomical variants. UUT obstruction in such settings calls for a variety of strategies with excellent outcomes.

Four hour voiding observation study in infants with posterior urethral valves.

INTRODUCTION AND AIM: Posterior urethral valves (PUV) are often associated with bladder dysfunction. Our primary aim was to investigate bladder status following primary valves resection to gather evidence of function and to guide early clinical management. PATIENTS AND METHODS: Between July 2015-2020, we prospectively evaluated bladder function of PUV infants. Primary outcomes measured were number of voids, voided volume (VV), post-void residual (PVR) and bladder capacity (BC). Statistical comparisons and descriptive analysis were carried out between groups using 2-tails T test and Chi square test using an IBM SPSS Statistics program version 25. RESULTS: Sixty-one infants were included. Median age at resection was 28 days (5 days-11 months). In thirty-eight patients (62%) diagnosis was suspected antenatally. Vesico-ureteric reflux (VUR) was present in 16 (26%). The 4-h observational study was performed at median 4 days following primary cold-knife valve resection and 1 day (range from 1 to 12 days) after catheter removal. Infants voided an average 6 times (2-13 voids). Bladder capacity was large in 20 (33%) and small in 4 (6.5%) patients with a median ratio BC/eBC = 1.2 (0.49-22.6). Median residual urine was 9 mls but with great variability among the cohort (0-121 mls). A big variability in bladder emptying was noted between patients and between voids performed in one patient. In 13 infants (21%) average PVR was larger than the average voided volume. In 26 (42.6%) PVR was larger than VV at least once. Thirty-eight infants (62.3%) emptied their bladders almost completely at least once (with PVR <5 mls). No difference was found in PVR, Median BC/EBC and PVR/BC between infants with and without VUR (p = 0.654, P: 0.594 and p = 00.481, respectively). DISCUSSION: We presented our experience of non-invasive bladder function assessment in infants affected by PUV following primary valve resection. An interesting data is the great variability identified both between patients and between voids performed in a single patient. Average number of voids was similar to the voiding pattern in healthy newborns what reported by Gladh but variability was greater. We observed up to 13 voids in some infants and we can speculate this could reflect the presence of an overactive bladder. We observed at least 1 complete emptying in only 62% of infants and 21% of the cohort had PVR bigger than VV. In almost half of the infants (42.6%) PVR was larger than VV at least once. CONCLUSIONS: Around 40% of infants affected by PUV have abnormal bladder capacity and almost half of them have significant post void residuals following primary resection. Although controversies and limitations are present, we believe that this non-invasive study can provide valuable information to understand the dynamic of the bladder, particularly in children affected by PUV and allow early intervention in children considered "at risk". Having a non invasive way of assessing can help tailor intervention and be useful for future research into early bladder intervention and improving outcomes.