Adoptive cell therapy for high grade gliomas using simultaneous temozolomide and intracranial mgmt-modified γδ t cells following standard post-resection chemotherapy and radiotherapy: current strategy and future directions.

Cellular therapies, including chimeric antigen receptor T cell therapies (CAR-T), while generally successful in hematologic malignancies, face substantial challenges against solid tumors such as glioblastoma (GBM) due to rapid growth, antigen heterogeneity, and inadequate depth of response to cytoreductive and immune therapies, We have previously shown that GBM constitutively express stress associated NKG2D ligands (NKG2DL) recognized by gamma delta (γδ) T cells, a minor lymphocyte subset that innately recognize target molecules via the γδ T cell receptor (TCR), NKG2D, and multiple other mechanisms. Given that NKG2DL expression is often insufficient on GBM cells to elicit a meaningful response to γδ T cell immunotherapy, we then demonstrated that NKG2DL expression can be transiently upregulated by activation of the DNA damage response (DDR) pathway using alkylating agents such as Temozolomide (TMZ). TMZ, however, is also toxic to γδ T cells. Using a p140K/MGMT lentivector, which confers resistance to TMZ by expression of O(6)-methylguanine-DNA-methyltransferase (MGMT), we genetically engineered γδ T cells that maintain full effector function in the presence of therapeutic doses of TMZ. We then validated a therapeutic system that we termed Drug Resistance Immunotherapy (DRI) that combines a standard regimen of TMZ concomitantly with simultaneous intracranial infusion of TMZ-resistant γδ T cells in a first-in-human Phase I clinical trial (NCT04165941). This manuscript will discuss DRI as a rational therapeutic approach to newly diagnosed GBM and the importance of repeated administration of DRI in combination with the standard-of-care Stupp regimen in patients with stable minimal residual disease.

Sacituzumab govitecan, a Trop-2-directed antibody-drug conjugate, for patients with epithelial cancer: final safety and efficacy results from the phase I/II IMMU-132-01 basket trial.

BACKGROUND: Sacituzumab govitecan (SG), a trophoblast cell surface antigen-2 (Trop-2)-directed antibody-drug conjugate, has demonstrated antitumor efficacy and acceptable tolerability in a phase I/II multicenter trial (NCT01631552) in patients with advanced epithelial cancers. This report summarizes the safety data from the overall safety population (OSP) and efficacy data, including additional disease cohorts not published previously. PATIENTS AND METHODS: Patients with refractory metastatic epithelial cancers received intravenous SG (8, 10, 12, or 18 mg/kg) on days 1 and 8 of 21-day cycles until disease progression or unacceptable toxicity. Endpoints for the OSP included safety and pharmacokinetic parameters with investigator-evaluated objective response rate (ORR per RECIST 1.1), duration of response, clinical benefit rate, progression-free survival, and overall survival evaluated for cohorts (n > 10 patients) of small-cell lung, colorectal, esophageal, endometrial, pancreatic ductal adenocarcinoma, and castrate-resistant prostate cancer. RESULTS: In the OSP (n = 495, median age 61 years, 68% female; UGT1A1∗28 homozygous, n = 46; 9.3%), 41 (8.3%) permanently discontinued treatment due to adverse events (AEs). Most common treatment-related AEs were nausea (62.6%), diarrhea (56.2%), fatigue (48.3%), alopecia (40.4%), and neutropenia (57.8%). Most common treatment-related serious AEs (n = 75; 15.2%) were febrile neutropenia (4.0%) and diarrhea (2.8%). Grade ≥3 neutropenia and febrile neutropenia occurred in 42.4% and 5.3% of patients, respectively. Neutropenia (all grades) was numerically more frequent in UGT1A1∗28 homozygotes (28/46; 60.9%) than heterozygotes (69/180; 38.3%) or UGT1A1∗1 wild type (59/177; 33.3%). There was one treatment-related death due to an AE of aspiration pneumonia. Partial responses were seen in endometrial cancer (4/18, 22.2% ORR) and small-cell lung cancer (11/62, 17.7% ORR), and one castrate-resistant prostate cancer patient had a complete response (n = 1/11; 9.1% ORR). CONCLUSIONS: SG demonstrated a toxicity profile consistent with previous published reports. Efficacy was seen in several cancer cohorts, which validates Trop-2 as a broad target in solid tumors.

Sacituzumab govitecan in previously treated hormone receptor-positive/HER2-negative metastatic breast cancer: final results from a phase I/II, single-arm, basket trial.

BACKGROUND: Trophoblast cell-surface antigen-2 (Trop-2) is expressed in epithelial cancers, including hormone receptor-positive (HR+) metastatic breast cancer (mBC). Sacituzumab govitecan (SG; Trodelvy®) is an antibody-drug conjugate composed of a humanized anti-Trop-2 monoclonal antibody coupled to SN-38 at a high drug-to-antibody ratio via a unique hydrolyzable linker that delivers SN-38 intracellularly and in the tumor microenvironment. SG was granted accelerated FDA approval for metastatic triple-negative BC treatment in April 2020. PATIENTS AND METHODS: We analyzed a prespecified subpopulation of patients with HR+/human epidermal growth factor receptor 2-negative (HER2-) HR+/HER2- mBC from the phase I/II, single-arm trial (NCT01631552), who received intravenous SG (10 mg/kg) and whose disease progressed on endocrine-based therapy and at least one prior chemotherapy for mBC. End points included objective response rate (ORR; RECIST version 1.1) assessed locally, duration of response (DOR), clinical benefit rate, progression-free survival (PFS), overall survival (OS), and safety. RESULTS: Fifty-four women were enrolled between 13 February 2015 and 1 June 2017. Median (range) age was 54 (33-79) years and all received at least two prior lines of therapy for mBC. At data cut-off (1 March 2019), 12 patients were still alive. Key grade ≥3 treatment-related toxicities included neutropenia (50.0%), anemia (11.1%), and diarrhea (7.4%). Two patients discontinued treatment due to treatment-related adverse events. No treatment-related deaths occurred. At a median follow-up of 11.5 months, the ORR was 31.5% [95% confidence interval (CI), 19.5%-45.6%; 17 partial responses]; median DOR was 8.7 months (95% CI 3.7-12.7), median PFS was 5.5 months (95% CI 3.6-7.6), and median OS was 12 months (95% CI 9.0-18.2). CONCLUSIONS: SG shows encouraging activity in patients with pretreated HR+/HER2- mBC and a predictable, manageable safety profile. Further evaluation in a randomized phase III trial (TROPiCS-02) is ongoing (NCT03901339). TRIAL REGISTRATION: ClinicalTrials.gov NCT01631552; https://clinicaltrials.gov/ct2/show/NCT01631552.

Modeling of gas transmission properties of polymeric films used for MA packaging of fruits.

High value fruits namely, apple (cv. Royal Delicious), guava (cv. Baruipur) and litchi (cv. Shahi) harvested at their commercial maturity were considered for MA packaging to enhance storage life. Polymeric films namely LDPE, BOPP, PVC, PVDC of different thickness were used for MA packaging study and various film characteristics such as gas transmission rates, water vapour transmission rate, clarity, strength and durability were evaluated. Mathematical model was developed based on Arrhenius type equation to predict gas transmission rate (GTR) and the developed model was found to be very good fit with the mean relative deviation modulus value quite less than 10 %. The GTR of the films increased with the increase in storage temperature and the magnitude of the increase varied with the film type and thickness. Regression models have been suitably developed to predict the oxygen transmission rate and carbon dioxide transmission rate of selected polymeric films and combined film laminates as a function of temperatures. Since, none of the individual films could meet the gas transmission requirements of MAP for selected fruits, two different films were tailored to form laminates that sufficed the requirements for prolonged storage with maintaining original quality.

Risk assessment for anterior cruciate ligament injury.

INTRODUCTION: Anterior cruciate ligament tears are one of the most frequent soft tissue injuries of the knee. A torn anterior cruciate ligament leaves the knee joint unstable and at risk for further damage to other soft tissues manifested as pain, dislocation, and osteoarthritis. A better understanding of the anatomical details of knee joints suffering anterior cruciate ligament tears is needed to understand and develop prediction models for anterior cruciate ligament injury and/or tear. MATERIALS AND METHODS: Magnetic resonance images of 32 patients with anterior cruciate ligament tears and 40 patients with non-tears were evaluated from a physician group practice. Digital measurements of femoral condyle length, femoral notch width, anterior cruciate ligament width in the frontal and sagittal plane, and the anterior cruciate ligament length in the sagittal plane were taken in both groups to identify trends. Monte Carlo simulations were performed (n = 2000) to evaluate the relationship between notch width index and sagittal width and to establish functional relationships among the anatomical parameters for potential injury risk. Sensitivity analysis performed shows the risk of anterior cruciate ligament injury a function of force and notch width index. RESULTS: Females have a significantly shorter anterior cruciate ligament when compared to that of males. The notch width index was also significantly different between torn and non-torn individuals. The NWI was not significantly different between genders (p value = 0.40). CONCLUSIONS: Anterior cruciate ligament injury has been shown to be caused by the forces which act on the ligament. These forces can result from hyperextension of the tibia or the internal rotation of tibia. The anatomical parameters of the knee joint (i.e., notch width index, anterior cruciate ligament width and length) have no role in the cause of an injury.

Characterization of excretory-secretory antigens of adult Toxocara canis by western blotting.

Toxocara canis is one of the most common helminth worm of dogs which continues to stimulate both public health concern alongside the higher scientific interest. It may cause visceral and ocular damage in humans especially in children. The identification of specific antigens of T. canis is important so as to develop better diagnostic techniques. Excretory-secretory (ES) antigens were prepared by culturing the adult T. canis worms in RPMI 1640 medium without serum supplementation followed by ammonium sulphate precipitation. These antigens were separated using sodium dodecyl sulphate-electrophoresis (SDS-PAGE). Recovered proteins ranged from 30 to 384 kDa. The specific reactivity of the T. canis excretory-secretory (TC-ES) proteins was checked by western blotting. The immuno-reactivity of the naturally infected dog sera with the TC-ES antigens showed five bands at 43, 57,105, 139 and 175 kDa. The immuno-reactivity of the hyper immune serum raised in rabbits against TC-ES antigens was observed with ten polypeptides of 21, 25, 30, 37, 45, 50, 57, 69, 77 and 105 kDa. Common antigens band were observed at 57 and 105 KDa. These antigens merit further evaluation as candidate for use in diagnosis of toxocariasis in humans and adult dogs.

Piper nigrum and piperine: an update.

Black pepper (Piper nigrum L.) is a very widely used spice, known for its pungent constituent piperine. However, in addition to its culinary uses, pepper has important medicinal and preservative properties, and, more recently, piperine has been shown to have fundamental effects on p-glycoprotein and many enzyme systems, leading to biotransformative effects including chemoprevention, detoxification, and enhancement of the absorption and bioavailability of herbal and conventional drugs. Based on modern cell, animal, and human studies, piperine has been found to have immunomodulatory, anti-oxidant, anti-asthmatic, anti-carcinogenic, anti-inflammatory, anti-ulcer, and anti-amoebic properties. In this review, the chemical constituents, biological activities, effects of processing, and future potential of black pepper and piperine have been discussed thoroughly.

A prospective multi-centre randomised controlled trial comparing PlasmaKnife with bipolar dissection tonsillectomy: evaluating an emerging technology.

OBJECTIVE: To clinically evaluate and compare the PlasmaKnife to bipolar electrocautery in paediatric tonsillectomy. METHODS: A prospective, multi-centred, double-blinded randomised controlled trial, conducted in central London teaching hospitals. The participants were 100 patients aged 2-16 years with recurrent tonsillitis or obstructive adenotonsillar hypertrophy awaiting a tonsillectomy were recruited in to the study. The primary outcome measures were throat, ear and swallowing pain scores over two weeks. Secondary outcome measures were return to normal diet, return to normal activity, analgesic requirements, operation time and intra-operative blood loss. RESULTS: Surgical dissection was similar between the two groups with minimal blood loss and comparable overall operative times. We found that PlasmaKnife tonsillectomy caused more throat pain at 24 h (p=0.02). There was a tendency for a higher proportion in the bipolar dissection group to return to a normal diet, at day 3 (p=0.05) and at day 7 (p=0.04). The bipolar dissection returned to normal activities in a larger proportion than the PlasmaKnife group at day 3 (p=0.02) and at day 7 (p=0.01). There is some evidence of an association between use of analgesia at day 14 and method of tonsillectomy (p=0.03); the PlasmaKnife group tended to use less analgesia. During the study, four secondary bleeds occurred in the PlasmaKnife group and one in the bipolar dissection group, and all were managed conservatively. CONCLUSION: Our study has found no significant advantage to PlasmaKnife over bipolar diathermy tonsillectomy. However, this preliminary study finds PlasmaKnife to be an interesting instrument and may warrant a larger randomised study to evaluate the potential advantages.

Escheriosomes entrapped DNA vaccine co-expressing Cu-Zn superoxide dismutase and IL-18 confers protection against Brucella abortus.

In the present study, we evaluated prophylactic prospective of liposome based DNA vaccine co-expressing Cu-Zn superoxide dismutase (SOD) along with interleukin-18 (IL-18) against experimental murine brucellosis. The immunization schedule involves liposome-mediated delivery of pVsod (encoding SOD of Brucella abortus) and pVIL18-sod (encoding IL-18 of mouse and SOD of B. abortus) DNA constructs. The data highlight potential of Escherichia coli lipid liposome (escheriosome) based DNA delivery vehicle to induce SOD specific humoral and cellular immune responses in the immunized mice. The co-expression of SOD along with IL-18 ensued in higher lymphoproliferative response and IFN-gamma production in comparison to the group of animals that were immunized with free form of SOD-DNA. Antibody response developed upon immunization with both DNA vaccines was of IgG2a type mainly. The results of the present study show that co-expression of IL-18 along with SOD polarized the antigen specific immune responses toward Th-1 direction, a desirable feature to control intracellular pathogens.

Death potentially secondary to sub-Tenon's block.

An 82-year-old ASA 2 patient underwent routine sub-Tenon's block for cataract surgery. One minute after injection of the local anaesthetic, the patient had a generalised tonic-clonic seizure and developed refractory ventricular fibrillation; subsequent resuscitation was unsuccessful. With no evidence for intravascular injection, the lack of structural brain abnormalities, and the most striking feature on post mortem examination being severe triple vessel coronary artery disease, it was concluded that this was primarily cardiac in origin; however, the possibility of brainstem anaesthesia should also be considered.

Mycobacterium bovis AN5 antigens vary in their ability to induce nitric oxide production in blood monocytes of experimentally infected cattle.

Reactive nitrogen intermediates (RNI) are the principal effector molecules of activated monocyte/macrophage populations, responsible for killing and inhibiting the growth of virulent mycobacteria. In vitro nitrite production by blood monocytes of cattle inoculated with live Mycobacterium bovis AN5 was assessed from 0 day through 45 weeks post inoculation (PI). High in vitro nitrite production was observed at the 8th and 12th weeks PI in sensitized cattle but reactivity had fallen by the 20th week PI. To assess the in vitro nitrite producing ability of monocytes induced by individual polypeptides within culture filtrate antigens (CFA) of M. bovis AN5, cellular blotting was performed using peripheral blood mononuclear cells (PBMC) at the 12th week PI. It was observed that polypeptides of MW 70, 65, 60, 25, 24 and 22 kDa of CFA induced high nitrite production by blood monocytes while many polypeptides had little or no effect.

Avian reovirus induces an inhibitory effect on lymphoproliferation in chickens.

The cellular immune responses of chickens inoculated with the vaccine strain S-1133 and/or a field isolate VA-1 of avian reovirus (ARV) were studied. Both strains of virus caused inhibition of the phytohaemagglutinin (PHA)-induced lymphoproliferative response of peripheral blood mononuclear cells (PBMC) and splenic mononuclear cells (SMC) during the initial stage from day 4 up to day 10 post-inoculation (PI), with a later return to the normal value. The inhibition in the PHA-induced lymphoproliferation of SMC could be partially overcome by depletion of adherent cells. The supernatant of the PHA-stimulated SMC culture was also checked in vitro for the presence of suppressive factor(s) produced in response to ARV infection. The culture supernatant from chickens at day 5 PI caused significant inhibition of the PHA-induced lymphoproliferation of control birds, suggesting the presence of suppressive factor(s). ARV infection also significantly inhibited IL-2 production on day 5. There was a significant increase in nitric oxide production by the splenic mononuclear cells of chickens inoculated with either strain of ARV.

Understanding the multiple functions of Nramp1.

Nramp1 regulates macrophage activation in infectious and autoimmune diseases. Nramp2 controls anaemia. Both are divalent cation (Fe(2+), Zn(2+), and Mn(2+)) transporters; Nramp2 a symporter of H(+) and metal ions, Nramp1 a H(+)/divalent cation antiporter. This provides a model for metal ion homeostasis in macrophages. Nramp2, localised to early endosomes, delivers extracellularly acquired divalent cations into the cytosol. Nramp1, localised to late endosomes/lysosomes, delivers divalent cations from the cytosol to phagolysosomes. Here, Fe(2+) generates antimicrobial hydroxyl radicals via the Fenton reaction. Zn(2+) and Mn(2+) may also influence endosomal metalloprotease activity and phagolysosome fusion. The many cellular functions dependent on metal ions as cofactors may explain the multiple pleiotropic effects of Nramp1, and its complex roles in infectious and autoimmune disease.

Protective role of adenine nucleotide translocase in O2-deficient hearts.

At subsaturating concentrations of palmitoyl-CoA, the carnitine-dependent oxidation of the palmitoyl portion by uncoupled rat heart mitochondria was stimulated by ADP or ATP. This effect was traced to the prevention of acyl-CoA binding to adenine nucleotide translocase and the consequent sparing of acyl-CoA for acylcarnitine formation. Palmitoyl-CoA oxidation was stimulated by ITP also although ITP served neither as a transportable substrate nor as an inhibitor of ADP transport. ITP and other nontransportable nucleoside di(tri)phosphates prevented octanoyl-CoA binding to mitochondria. ITP was bound to mitochondria, and this binding was reversed by ADP, octanoyl-CoA, and carboxyatractyloside. Thus, besides a substrate site, there is a site on the translocase that binds nucleoside di(tri)phosphates, CoA and its esters, and atractylosides; inhibition of the translocase results, however, only from the binding of CoA esters of fatty acids and of atractylosides. We suggest that in O2-deficient hearts, when nucleotides decline and fatty acyl-CoA rises, the binding of the latter to the translocase becomes operational to slow fatty acylcarnitine production. By retarding the rise in amphipathic burden, this mechanism could protect heart against irreversible damage during brief periods of ischemia or hypoxia.

Radioisotopic assay of femtomole quantities of total adenine nucleotides, ATP plus ADP, and AMP.

AMP is converted to ATP by incubating overnight with pyruvate kinase, phosphoenolpyruvate and adenylate kinase in the presence of endogenous ATP (ADP) as primer. In a subsequent incubation in the presence of pyruvate kinase, phosphoenolpyruvate, radioactive glucose and hexokinase, ATP and ADP are estimated together by coupling their recycling to the formation of glucose 6-phosphate. The latter is separated by precipitation using 76% (v/v) acetone for radioactivity measurement in the same Eppendorf tube. The sensitivity of these simple procedures matches or exceeds those of luciferase methods of nucleotide determination.

Inhibition of mitochondrial carnitine-acylcarnitine translocase by sulfobetaines.

Sulfobetaines (N-alkyl-N,N-dimethyl-3-ammonio-1-propanesulfonates) have been identified as relatively specific and selective inhibitors of mitochondrial carnitine-acylcarnitine translocase. Thus, sublytic concentrations of sulfobetaines (alkyl = octyl to tetradecyl) inhibit the respiration of rat heart mitochondria supported by added acylcarnitines or pyruvate plus malonate and carnitine. Both exchange efflux and unidirectional net efflux of mitochondrial carnitine are also inhibited; the half-maximal inhibition of the former occurs at micromolar concentrations of sulfobetaines and the inhibitory effect is reversible and competitive with respect to carnitine. As a stop-inhibitor, 20 mM sulfobetaine, (alkyl = octyl), is useable at near 0 degrees C but is less effective than 2 mM mersalyl when transport rates are very rapid as at higher temperatures especially with liver mitochondria. The loss of mitochondrial carnitine that normally occurs owing to the progress of net efflux during the isolation of mitochondria is prevented by the inclusion of 20 mM sulfobetaine in the isolation medium and this enables a better estimate of the mitochondrial carnitine content. Sulfobetaines inhibit the activities of mitochondrial carnitine acetyltransferase and carnitine palmitoyltransferase but only at concentrations severalfold higher than those inhibitory for the translocase. This observation supports the belief that carnitine-acylcarnitine translocase is an entity distinct from that of carnitine acyltransferases.