Decrease In Cell Count Research Articles

The Tragedy of Alzheimer's Disease: Towards Better Management via Resveratrol-Loaded Oral Bilosomes.

Alzheimer’s disease (AD) is a neurodegenerative disease where oxidative stress plays a major role as a key pathologic factor. The study aims to develop resveratrol (RES)-loaded bilosomes for oral use, aiming to enhance RES bioavailability. RES-loaded bilosomes were prepared using the thin-film hydration technique. The effect of different formulation variables viz. the number of extrusion cycles, drug concentration and the effect of pH of the medium and cholesterol addition on the physicochemical properties of the prepared bilosomes was investigated. Results revealed the successful entrapment of RES into bilosomes. An optimized formula was selected, showing the lowest particle size (189 ± 2.14), acceptable PDI (0.116) and entrapment efficiency (76.2 ± 1.36). In vivo studies on a streptozotocin-induced animal model of AD showed the preeminence of bilosomes over traditional drug suspension to enhance mice memory via Y-maze and Morris water maze tests. Moreover, mice treated with the optimized formula exhibited decreased COX2, IL-6, amyloid-beta peptide and Tau protein levels compared to the drug suspension. Immuno-histochemical analysis revealed a significant decrease of glial fibrillary acidic protein values and microglial cell count in mice treated with bilosomes. Finally, it could be advocated that RES-loaded bilosomes could be a promising drug delivery system to control AD.

Alpha lipoic acid ameliorates detrimental effects of maternal lipopolysaccharides exposure on prefrontal white matter in adult male offspring rats

BackgroundActivation of the maternal immune system by lipopolysaccharide (LPS) increases the production of proinflammatory cytokines, free radicals, and reactive oxygen species (ROS), all of which play a significant role in the pathogenesis of many offspring neurodevelopmental disorders. Alpha Lipoic Acid (ALA) is a natural compound that has anti-inflammatory and antioxidant properties. This study was performed to assess the effect of prenatal exposure to LPS on the prefrontal white matter of rat offspring and evaluate the potential protective effects of ALA co-administration during pregnancy. MethodsPregnant Wistar rats were randomly divided into six groups (n = 6 each group): (1) control, (2) received LPS (100 μg/kg, intraperitoneally (IP) on gestational day 9.5 (GD 9.5), (3) received ALA (20 mg/kg) from GD1 to GD11, (4) LPS+ALA received LPS on GD9.5 and ALA from GD1 to GD11, (5 and 6) received LPS and ALA vehicle respectively. In each group, 21-day old male offspring (2 male pups from each mother) was harvested, and then their prefrontal white matter was separated and prepared for the ultrastructural, stereological, and molecular assays. ResultsIn utero exposure to LPS led to a significant decrease in nerve cell counts, ultrastructural alterations in myelinated axons, and abnormal changes in genes expression of Sox10,Olig1,yrf,Wnt in the prefrontal of the rat offspring. Co-administration of ALA resulted in amelioration of those abnormal changes in the LPS rat offspring. ConclusionThe findings of our preclinical study, explore that prenatal ALA treatment efficiently protects the nervous system against LPS induced abnormal changes in the offspring.

Гематологические особенности у больных ревматоидным артритом при эндопротезировании тазобедренного сустава

(THR). Inflammation in RA is the main factor manifesting anaemia, neutropenia, thrombocytosis and eosinophilia. The changes in blood components are important for the outcomes of major orthopaedic surgery. The purpose was to identify hematological parameters in RA patients undergoing THR and assess the effect on intraoperative blood loss. Material and methods Outcomes of 44 THR patients treated for grade III degenerative coxarthrosis (n = 21, OA group) and RA coxarthrosis (n = 23, RA group) in Ekaterinburg regional hospital № 1 between 2018 and 2019 were reviewed. The patients' age ranged from 41 to 70 years. Clinical, radiological, laboratory examinations, computed tomography and statistical analysis were used for the study. Cell counting was produced with the Sysmex XT-4000i automated hematology system. Statistical analysis was performed using the tools of Statistica software. Nonparametric Mann-Whitney test was used to compare cell counts between the groups. The Spearman Rank correlation was used to analyse the correlation between the the cell counts in the groups. For calculations, a significance level of р < 0.05 was adopted. Results There were no significant differences in the preoperative white blood cell (WBC), neutrophil, lymphocyte, monocyte, eosinophil counts between RA and OA groups. The RA group showed an evident decrease in red blood cell (RBC) count and haemoglobin level as compared to OA group. The RA group demonstrated the higher platelet count with mean platelet volume (MPV) being significantly lower than that in the OA group. WBC count, neutrophils, in particular, was shown to increase with lymphocyte, RBC, platelet count and hemoglobin, plateletcrit levels decreased at 24 hours postoperatively. There were no significant differences in WBC and RBC counts in the groups postoperatively. The differences in the MPV were leveled up in the groups with the platelet count being higher in the RA group as compared to the OA group. Conclusions Hematological parameters of RA patients who had undergone specific preoperative preparation were not shown to be associated with greater blood loss during hip replacement surgery. The leukocyte count leveled up in the preoperative and early postoperative periods can be indicative of the absence of a significant effect of RA on the postoperative inflammation.

Degradation of p-cresol, resorcinol, and phenol in anaerobic membrane bioreactors under saline conditions

Treating petrochemical wastewater is a challenge for conventional anaerobic reactors. One example is coal gasification wastewater that, besides its salinity, is rich in toxic and inhibitory aromatics, such as phenol, cresols, and resorcinol. Studies have shown that phenol and p-cresol share the same degradation intermediates, whereas resorcinol is degraded via another route. This study investigated the simultaneous degradation of p-cresol or resorcinol with phenol under anaerobic saline conditions. Batch experiments with anaerobic phenol-degrading biomass were conducted to assess the feasibility of the degradation of p-cresol and resorcinol. Volumetric uptake rates of 11.4 ± 2.4 mgp-cresol·L–1d–1 and 4.2 ± 1.9 mgresorcinol·L–1d–1 were determined. The effect of p-cresol and resorcinol on the specific methanogenic activity and the cell viability in phenol-degrading and non-adapted biomass was assessed. Half maximal inhibitory concentration (IC50) values of 0.73 gp-cresol·L-1 and 3.00 gresorcinol·L-1 were estimated for phenol-degrading biomass, whereas IC50 values of 0.60 gp-cresol·L-1 and 0.25 gresorcinol·L-1 were estimated for the non-adapted biomass. p-Cresol caused a higher decrease in the non-damaged cell counts in comparison to resorcinol. Two anaerobic membrane bioreactors under saline conditions [8 g Na+·L–1] were fed with mixtures of either phenol-p-cresol or phenol-resorcinol. At an influent phenol concentration of 2 g·L-1, maximum conversion rates of 22 mgp-cresol·gVSS-1d–1 and 16 mgresorcinol·gVSS–1d–1 were found. In both AnMBRs, Syntrophorhabdus sp. and Methanosaeta sp. were the most abundant bacteria and methanogen, respectively. The feasibility of simultaneous conversion of phenolic compounds under saline conditions in AnMBRs opens novel perspectives for the high-rate anaerobic treatment of chemical wastewater.

A phase II study of sequential treatment with anthracycline and taxane followed by eribulin in patients with HER2-negative, locally advanced breast cancer (JBCRG-17)

PurposeThe sequence of taxanes (T) followed by anthracyclines (A) as neoadjuvant chemotherapy has been the standard of care for almost 20 years for locally advanced breast cancer (LABC). Sequential administration of eribulin (E) following A/T could provide a greater response rate for women with LABC.MethodsIn this single-arm, multicenter, Phase II prospective study, the patients received 4 cycles of the FEC regimen and 4 cycles of taxane. After the A/T-regimen, 4 cycles of E were administered followed by surgical resection. The primary endpoint was the clinical response rate. Eligible patients were women aged 20 years or older, with histologically confirmed invasive breast cancer, clinical Stage IIIA (T2–3 and N2 only), Stage IIIB, and Stage IIIC, HER2-negative.ResultsA preplanned interim analysis aimed to validate the trial assumptions was conducted after treatment of 20 patients and demonstrated that clinical progressive disease rates in the E phase were significantly higher (30%) than assumed. Therefore, the Independent Data Monitoring Committee recommended stopping the study. Finally, 53 patients were enrolled, and 26 patients received the A/T/E-regimen. The overall observed clinical response rate (RR) was 73% (19/26); RRs were 77% (20/26) in the AT phase and 23% (6/26) in the E phase. Thirty percent (8/26) of patients had PD in the E phase, 6 of whom had achieved cCR/PR in the AT phase. Reported grade ≥ 3 AEs related to E were neutropenia (42%), white blood cell count decrease (27%), febrile neutropenia (7.6%), weight gain (3.8%), and weight loss (3.8%).ConclusionSequential administration of eribulin after the A/T-regimen provided no additional effect for LABC patients. Future research should continue to focus on identifying specific molecular biomarkers that can improve response rates.

CDPA-1 Stored Blood Induced Effect on some Haematological Parameters

Background: Most blood collection bags contain 63 mL CPDA anticoagulant which is sufficient to anticoagulant and ensures the viability of blood cells in 450 mL±10% blood for up to 28–35 days when the blood is stored at 2–8°C. Prolonged storage of blood leads to alteration in cells hematologically which may lose viability with time. Aim: The study was conducted to determine the effect of storage on CPDA-1 for varying periods on some hematological parameters. Materials and methods: The study was conducted on blood donated by 30 healthy volunteer donors. Effect of storage was analyzed at 1, 7, 14, 21and 28 days intervals. Hematological parameters were measured using Mindary PS 300 hematology analyzer. Results: There is a highly significant increase in hemoglobin concentration, packed cell volume (P.C.V. %), MCV, and also a decrease in lymphocyte, granulocyte, and platelet count. The results also showed an insignificant decrease in total white blood cell count. Conclusion: There are degenerative changes observed in blood parameters in samples collected in citrate phosphate dextrose adenine (CPDA-1).

A comparative study of divide and conquer, stop and chop and phaco chop techniques of nucleotomy in phacoemulsification

Phacoemulsification is a variant of extracapsular cataract extraction surgery which is the gold standard treatment modality for cataract surgery nowadays. The common nucleotomy techniques of phacoemulsification are divide and conquer, stop and chop and phaco chop.Aim of this studyto compare different techniques of nucleotomy during phacoemulsification with respect to corneal endothelial cell loss, central corneal thickness (CCT) and comment on their merits and demerits. 60 patients undergoing cataract surgery by phacoemulsification were randomly divided into three groups of 20 each (Group A, B and C) who underwent nucleotomy by divide and conquer, stop and chop and phaco chop technique respectively. The preoperative endothelial cell density and central corneal thickness were recorded using Topcon SP3000P non-contrast specular microscope. All patients were followed up at 1, 4 and 12week after surgery and evaluated for vision, slit lamp examination, endothelial cell count and central corneal thickness.The age group of patients studied was 50-80 years, where maximum number of patients were in 51-60 years age group. Out of 60 patients, most common morphological type of cataract was nuclear (40%) cataract followed by cortico-nuclear (36.67%) cataract. After 1 week of surgery, drop in endothelial cell density was 7.06% in Group A, 6.49% in Group B and 6.84% in Group C patients. A further slight decrease in endothelial cell count was recorded during 4week and on final follow up visit at 12 weeks after cataract surgery, there was an endothelial cell loss of 9.51%, 10.9% and 11% in Group A, B and C respectively. The central corneal thickness at 1postoperative week increased by 2.75%, 2.35% and 2.20% in group A, B and C respectively. Further evaluation of CCT at 4and 12 week after surgery showed the values to be almost same as preoperative values. The rate of complications, which mainly included postoperative keratitis, mild anterior uveitis and cystoid macular edema, were found to be almost the same in all the three study groups.All the three nucleotomy techniques of phacoemulsification i.e divide and conquer, stop and chop and phaco chop are equally efficacious in terms of CCT changes, endothelial cell loss and complications rate in the hands of an experienced surgeon.

The Adaptive Immune Landscape of the Colorectal Adenoma-Carcinoma Sequence.

Background. The tumor immune microenvironment exerts a pivotal influence in tumor initiation and progression. The aim of this study was to analyze the immune context of sporadic and familial adenomatous polyposis (FAP) lesions along the colorectal adenoma–carcinoma sequence (ACS). Methods. We analyzed immune cell counts (CD3+, CD4+, CD8+, Foxp3+, and CD57+), tumor mutation burden (TMB), MHC-I expression and PD-L1 expression of 59 FAP and 74 sporadic colorectal lesions, encompassing adenomas with low-grade dysplasia (LGD) (30 FAP; 30 sporadic), adenomas with high-grade dysplasia (22 FAP; 30 sporadic), and invasive adenocarcinomas (7 FAP; 14 sporadic). Results. The sporadic colorectal ACS was characterized by (1) a stepwise decrease in immune cell counts, (2) an increase in TMB and MHC-I expression, and (3) a lower PD-L1 expression. In FAP lesions, we observed the same patterns, except for an increase in TMB along the ACS. FAP LGD lesions harbored lower Foxp3+ T cell counts than sporadic LGD lesions. A decrease in PD-L1 expression occurred earlier in FAP lesions compared to sporadic ones. Conclusions. The colorectal ACS is characterized by a progressive loss of adaptive immune infiltrate and by the establishment of a progressively immune cold microenvironment. These changes do not appear to be related with the loss of immunogenicity of tumor cells, or to the onset of an immunosuppressive tumor microenvironment.

Environmentally Relevant Levels of Depleted Uranium Impacts Dermal Fibroblast Proliferation, Viability, Metabolic Activity, and Scratch Closure.

Uranium (U) is a heavy metal used in military and industrial settings, with a large portion being mined from the Southwest region of the United States. Uranium has uses in energy and military weaponry, but the mining process has released U into soil and surface waters that may pose threats to human and environmental health. The majority of literature regarding U’s human health concern focuses on outcomes based on unintentional ingestion or inhalation, and limited data are available about its influence via cutaneous contact. Utilizing skin dermis cells, we evaluated U’s topical chemotoxicity. Employing soluble depleted uranium (DU) in the form of uranyl nitrate (UN), we hypothesized that in vitro exposure of UN will have cytotoxic effects on primary dermal fibroblasts by affecting cell viability and metabolic activity and, further, may delay wound healing aspects via altering cell proliferation and migration. Using environmentally relevant levels of U found in water (0.1 M to 100 M [UN]; 23.8–23,800 ppb [U]), we quantified cellular mitosis and migration through growth curves and in vitro scratch assays. Cells were exposed from 24 h to 144 h for a time-course evaluation of UN chemical toxicity. The effects of UN were observed at concentrations above and below the Environmental Protection Agency threshold for safe exposure limits. UN exposure resulted in a dose-dependent decrease in the viable cell count; however, it produced an increase in metabolism when corrected for the viable cells present. Furthermore, cellular proliferation, population doubling, and percent closure was hindered at levels ≥10 M UN. Therefore, inadvertent exposure may exacerbate pre-existing skin diseases in at-risk demographics, and additionally, it may substantially interfere in cutaneous tissue repair processes.

Evaluation of Mast Cells in Oral Potentially Malignant Disorders and Oral Squamous Cell Carcinoma.

Background Oral squamous cell carcinoma (OSCC) and oral potentially malignant disorders (OPMDs) are epithelial-derived pathologies which share inflammation as a common initial pathogenic-inducing state. Mast cell is a key immune modulating cell which is primarily involved in initiation and propagation of inflammation. The role of mast cell in OPMDs and OSCC has been an established fact; however, its definitive pathogenic correlation is still under study. The objective of the study was to evaluate the number of mast cell in OPMDs and OSCC using special stain correlating its probable role as a promoter or retarder of OSCC. Materials and Methods Forty-five archival histopathologically confirmed cases each of OPMD and OSCC were studied for mast cells using toluidine blue and Alcian blue-safranin stain and compared with 10 normal oral mucosal tissues. Comparisons between the mast cells count was also performed between the two special stains. Results Among 100 cases, 67% were males and 33% were females. The mean age was 41.68 ± 13.39; 55.06 ± 12.55; and 18.4 ± 2.54 years for OPMDs, OSCC, and normal, respectively. A statistically significant increase in mast cells among OPMDs (9.88 ± 7.9) and OSCC (6.711 ± 3.94) was observed compared to normal oral mucosa. The mast cell count reduced among OSCC in comparison to OPMDs which was significant as well. The mean mast cell count for Alcian blue-safranin stain was higher than toluidine blue stain. Conclusion There is a decrease in mean mast cell count from OPMDs to OSCC which is suggestive of protective role of mast cell. Proper quantification of mast cells using specific stains can guide to define prognosis in oral potentially malignant disorders and oral cancer.

One Year Outcomes and Stability of a Novel Scleral Anchored Intraocular Lens.

Purpose To assess one year results and stability of the implantation of a scleral anchored intraocular lens (IOL). Design Interventional prospective case series. Methods Sixty eyes of 60 patients affected by either aphakia or IOL dislocation were included in this study. Patients underwent vitrectomy, scleral fixation of the IOL, and, if present, dislocated IOL removal. Patients were evaluated preoperatively and at 1, 3, 6, and 12 months after surgery by best-corrected distance visual acuity (BCVA) assessment, intraocular pressure (IOP) measurement, corneal specular microscopy, and optical coherence tomography (OCT) of both the macula and anterior segment. Results At twelve months, mean BCVA significantly improved (p < 0.0001), and none of the patients experienced a decrease of visual acuity. A 10% decrease of endothelial cell count occurred after surgery. Cystoid macular edema occurred in three patients (5%). A transient increase of intraocular pressure was noted in 7 cases (12%). At one month, horizontal and vertical IOL tilt was 1.04 ± 0.87 and 0.74 ± 0.71 degrees, respectively, and did not significantly change in the follow-up (p > 0.05). None of the patients had decentration or dislocation of scleral-fixated IOL during the follow-up. Conclusion Implantations of scleral plug fixated IOL provide good visual results, low complication rate, and excellent stability of the lens until one-year follow-up.

NOTCH-mediated exvivo expansion of human hematopoietic stem and progenitor cells by culture under hypoxia.

SummaryActivation of NOTCH signaling in human hematopoietic stem/progenitor cells (HSPCs) by treatment with an engineered Delta-like ligand (DELTA1ext-IgG [DXI]) has enabled ex vivo expansion of short-term HSPCs, but the effect on long-term repopulating hematopoietic stem cells (LTR-HSCs) remains uncertain. Here, we demonstrate that ex vivo culture of human adult HSPCs with DXI under low oxygen tension limits ER stress in LTR-HSCs and lineage-committed progenitors compared with normoxic cultures. A distinct HSC gene signature was upregulated in cells cultured with DXI in hypoxia and, after 21 days of culture, the frequency of LTR-HSCs increased 4.9-fold relative to uncultured cells and 4.2-fold compared with the normoxia + DXI group. NOTCH and hypoxia pathways intersected to maintain undifferentiated phenotypes in cultured HSPCs. Our work underscores the importance of mitigating ER stress perturbations to preserve functional LTR-HSCs in extended cultures and offers a clinically feasible platform for the expansion of human HSPCs.

Metagenomic Next-Generation Sequencing of Bloodstream Microbial Cell-Free Nucleic Acid in Children With Suspected Sepsis in Pediatric Intensive Care Unit.

Bloodstream infection is a life-threatening complication in critically ill patients. Multi-drug resistant bacteria or fungi may increase the risk of invasive infections in hospitalized children and are difficult to treat in intensive care units. The purpose of this study was to use metagenomic next-generation sequencing (mNGS) to understand the bloodstream microbiomes of children with suspected sepsis in a pediatric intensive care unit (PICU). mNGS were performed on microbial cell-free nucleic acid from 34 children admitted to PICU, and potentially pathogenic microbes were identified. The associations of serological inflammation indicators, lymphocyte subpopulations, and other clinical phenotypes were also examined. mNGS of blood samples from children in PICU revealed potential eukaryotic microbial pathogens. The abundance of Pneumocystis jirovecii was positively correlated with a decrease in total white blood cell count and immunodeficiency. Hospital-acquired pneumonia patients showed a significant increase in blood bacterial species richness compared with community-acquired pneumonia children. The abundance of bloodstream bacteria was positively correlated with serum procalcitonin level. Microbial genome sequences from potential pathogens were detected in the bloodstream of children with suspected sepsis in PICU, suggesting the presence of bloodstream infections in these children.

Markov modelling of viral load adjusting for CD4 orthogonal variable and multivariate conditional autoregressive mapping of the HIV immunological outcomes among ART patients in Zimbabwe

BackgroundThis study aimed to jointly model HIV disease progression patterns based on viral load (VL) among adult ART patients adjusting for the time-varying “incremental transients states” variable, and the CD4 cell counts orthogonal variable in a single 5-stage time-homogenous multistate Markov model. We further jointly mapped the relative risks of HIV disease progression outcomes (detectable VL (VL ≥ 50copies/uL) and immune deterioration (CD4 < 350cells/uL) at the last observed visit) conditional not to have died or become loss to follow-up (LTFU).MethodsSecondary data analysis of individual-level patients on ART was performed. Adjusted transition intensities, hazard ratios (HR) and regression coefficients were estimated from the joint multistate model of VL and CD4 cell counts. The mortality and LTFU transition rates defined the extent of patients’ retention in care. Joint mapping of HIV disease progression outcomes after ART initiation was done using the Bayesian intrinsic Multivariate Conditional Autoregressive prior model.ResultsThe viral rebound from the undetectable state was 1.78times more likely compared to viral suppression among patients with VL ranging from 50-1000copies/uL. Patients with CD4 cell counts lower than expected had a higher risk of viral increase above 1000copies/uL and death if their VL was above 1000copies/uL (state 2 to 3 (λ23): HR = 1.83 and (λ34): HR = 1.42 respectively). Regarding the time-varying effects of CD4 cell counts on the VL transition rates, as the VL increased, (λ12 and λ23) the transition rates increased with a decrease in the CD4 cell counts over time. Regardless of the individual’s VL, the transition rates to become LTFU decreased with a decrease in CD4 cell counts. We observed a strong shared geographical pattern of 66% spatial correlation between the relative risks of detectable VL and immune deterioration after ART initiation, mainly in Matabeleland North.ConclusionWith high rates of viral rebound, interventions which encourage ART adherence and continual educational support on the barriers to ART uptake are crucial to achieve and sustain viral suppression to undetectable levels. Area-specific interventions which focus on early ART screening through self-testing, behavioural change campaigns and social support strategies should be strengthened in heavily burdened regions to sustain the undetectable VL. Sustaining undetectable VL lowers HIV transmission in the general population and this is a step towards achieving zero HIV incidences by 2030.

Rituximab induces rapid blood repopulation by CLL cells mediated through their release from immune niches and complement exhaustion

The in vivo rituximab effects in B cell malignancies are only partially understood. Here we analyzed in a large chronic lymphocytic leukemia (CLL) cohort (n = 80) the inter-patient variability in CLL cell count reduction within the first 24 h of rituximab administration in vivo, and a phenomenon of blood repopulation by malignant cells after anti-CD20 antibody therapy. Larger CLL cell elimination after rituximab infusion was associated with lower pre-therapy CLL cell counts, higher CD20 levels, and the non-exhausted capacity of complement-dependent cytotoxicity (CDC). The absolute amount of cell-surface CD20 molecules (CD20 density x CLL lymphocytosis) was a predictor for complement exhaustion during therapy. We also describe that a highly variable decrease in CLL cell counts at 5 h (88 %–2%) following rituximab infusion is accompanied in most patients by peripheral blood repopulation with CLL cells at 24 h, and in ∼20 % of patients, this resulted in CLL counts higher than before therapy. We provide evidence that CLL cells recrudescence is linked with i) CDC exhaustion, which leads to the formation of an insufficient amount of membrane attack complexes, likely resulting in temporary retention of surviving rituximab-opsonized cells by the mononuclear-phagocyte system (followed by their release back to blood), and ii) CLL cells regression from immune niches (CXCR4dimCD5bright intraclonal subpopulation). Patients with major peripheral blood CLL cell repopulation exhibited a longer time-to-progression after chemoimmunotherapy compared to patients with lower or no repopulation, suggesting chemotherapy vulnerability of CLL cells that repopulate the blood.

Serum selenium predicts achievement of full-dose cisplatin in concurrent chemoradiotherapy for locally advanced head and neck squamous cell carcinoma: A prospective, observational study

ObjectiveConcurrent chemoradiotherapy (CCRT) with three-weekly high-dose cisplatin (100 mg/m2) is a standard treatment for locally advanced head and neck squamous cell carcinoma (HNSCC), but compliance with cisplatin is often poor due to various adverse events. The aim of this prospective, observational study was to determine the predictors of achievement of full-dose cisplatin. MethodsA prospective, observational study was conducted involving 60 patients who received CCRT with three-weekly high-dose cisplatin (100 mg/m2) for locally advanced HNSCC. Possible predictors affecting compliance with cisplatin were subjected to univariate and multivariate logistic regression analyses. Age, sex, primary site, clinical stage, treatment intent, history of hypertension, diabetes mellitus, smoking and drinking habits, body mass index, creatinine clearance, serum albumin, controlling nutrition status, trace elements (Fe, Zn, Cu, Se), acute kidney injury, white blood cell count decrease, neutrophilia, and weight loss were the variables evaluated. ResultsTwenty-seven patients achieved full-dose cisplatin (300 mg/m2), and the other 33 patients did not. Multivariate logistic regression analysis showed that both mild renal dysfunction and selenium deficiency before treatment independently had negative impacts on achievement of full-dose cisplatin. ConclusionsAs well as renal function, selenium deficiency is a potential therapeutic target for CCRT with high-dose cisplatin in HNSCC patients.

RNA sequencing of whole blood reveals early alterations in immune cells and gene expression in Parkinson's disease.

Changes in the blood-based RNA transcriptome have the potential to inform biomarkers of Parkinson's disease (PD) progression. Here we sequenced a discovery set of whole-blood RNA species in 4,871 longitudinally collected samples from 1,570 clinically phenotyped individuals from the Parkinson's Progression Marker Initiative (PPMI) cohort. Samples were sequenced to an average of 100 million read pairs to create a high-quality transcriptome. Participants with PD in the PPMI had significantly altered RNA expression (>2,000 differentially expressed genes), including an early and persistent increase in neutrophil gene expression, with a concomitant decrease in lymphocyte cell counts. This was validated in a cohort from the Parkinson's Disease Biomarkers Program (PDBP) consisting of 1,599 participants and by alterations in immune cell subtypes. This publicly available transcriptomic dataset, coupled with available detailed clinical data, provides new insights into PD biological processes impacting whole blood and new paths for developing diagnostic and prognostic PD biomarkers.

Structural Changes in Thin Keratoconic Corneas Following Crosslinking with Hypotonic Riboflavin: Findings on In Vivo Confocal Microscopy.

PurposeTo report structural changes observable in in vivo confocal microscopy (IVCM) in keratoconic corneas 400 μm treated with hypotonic riboflavin and collagen crosslinking (CXL).MethodsTen eyes of ten patients with progressive keratoconus and corneal thickness between 350 and 399 μm underwent CXL with hypotonic riboflavin. IVCM was performed preoperatively and at one month, three months, and six months after the procedure.ResultsIVCM analysis one month postoperatively showed complete absence of the subepithelial nerve plexus with gradual regeneration over six months in 8 of the 10 eyes, and poor regeneration in the remaining 2 eyes. The anterior stroma showed extracellular lacunae and hyper-reflective cytoplasm in a honeycomb appearance signifying edema at one month which gradually decreased over six months post CXL. Stromal keratocyte apoptosis was evident in the anterior stroma in all cases and extended to the posterior stroma in four eyes with gradual regeneration evident at three and six months. The specular endothelial count decreased by 8% (P = 0.005) post-CXL, but no corneas developed clinical signs of endothelial trauma.ConclusionIVCM analysis of thin corneas after hypotonic CXL showed posterior corneal structural changes. Posterior stromal changes were accompanied by a decrease in the endothelial cell count. This case series was a preliminary feasibility study that might necessitate conducting a well-designed controlled study.

Nanoparticulate matter exposure results in white matter damage and an inflammatory microglial response in an experimental murine model.

Exposure to ambient air pollution has been associated with white matter damage and neurocognitive decline. However, the mechanisms of this injury are not well understood and remain largely uncharacterized in experimental models. Prior studies have shown that exposure to particulate matter (PM), a sub-fraction of air pollution, results in neuroinflammation, specifically the upregulation of inflammatory microglia. This study examines white matter and axonal injury, and characterizes microglial reactivity in the corpus callosum of mice exposed to 10 weeks (150 hours) of PM. Nanoscale particulate matter (nPM, aerodynamic diameter ≤200 nm) consisting primarily of traffic-related emissions was collected from an urban area in Los Angeles. Male C57BL/6J mice were exposed to either re-aerosolized nPM or filtered air for 5 hours/day, 3 days/week, for 10 weeks (150 hours; n = 18/group). Microglia were characterized by immunohistochemical double staining of ionized calcium-binding protein-1 (Iba-1) with inducible nitric oxide synthase (iNOS) to identify pro-inflammatory cells, and Iba-1 with arginase-1 (Arg) to identify anti-inflammatory/ homeostatic cells. Myelin injury was assessed by degraded myelin basic protein (dMBP). Oligodendrocyte cell counts were evaluated by oligodendrocyte transcription factor 2 (Olig2). Axonal injury was assessed by axonal neurofilament marker SMI-312. iNOS-expressing microglia were significantly increased in the corpus callosum of mice exposed to nPM when compared to those exposed to filtered air (2.2 fold increase; p<0.05). This was accompanied by an increase in dMBP (1.4 fold increase; p<0.05) immunofluorescent density, a decrease in oligodendrocyte cell counts (1.16 fold decrease; p<0.05), and a decrease in neurofilament SMI-312 (1.13 fold decrease; p<0.05) immunofluorescent density. Exposure to nPM results in increased inflammatory microglia, white matter injury, and axonal degradation in the corpus callosum of adult male mice. iNOS-expressing microglia release cytokines and reactive oxygen/ nitrogen species which may further contribute to the white matter damage observed in this model.

Abstract 1007: Patient-derived metastatic renal carcinoma cells are highly sensitive to in vitro Tumor Treating Fields (TTFields)

Abstract Background: Until the advent of checkpoint inhibitor (CI) therapies, renal cell carcinoma (RCC) was clinically challenging to treat given its radio- and chemotherapy resistance. Despite the advances with CIs, patients with brain metastases derived from RCC still face challenges with treatment due to limited brain penetration of CI compounds. Anti-mitotic TTFields therapy is FDA-approved for the treatment of glioblastoma and mesothelioma, and clinical trials are underway for the application of TTFields therapy to primary lung cancer as well as lung cancer brain metastases. In this study, in vitro TTFields were applied to patient-derived brain metastasis RCC cells to determine the potential efficacy of this emerging treatment modality. Methods: This study was approved by the Institutional Review Board. A single-cell suspension was generated from tumor tissues obtained from a patient with newly-diagnosed RCC brain metastasis (male, age 52, mutant p53). Cells were cultured in DMEM/F12 media with 10% fetal bovine serum and gentamicin. Prior to TTFields initiation, cells were plated on plastic coverslips (1×104cells/coverslip; n=8/group) and incubated overnight. Then, TTFields were applied at 200 kHz with a field intensity of ~1.6 V/cm for 14 days, with one group of coverslips maintained as untreated controls. At the conclusion of the experiment, cells were harvested, stained with acridine orange/propidium iodide and counted with a Cellometer K2 (Nexcelom Bioscience, Lawrence, MA) that provides viable cell counts and average cell diameter. Groups were compared with a two-tailed t-test (GraphPad Prism v9, GraphPad Software, San Diego, CA) with significance set at P&amp;lt;0.05. Results: Viable cell count per coverslip for the control group was 149,863 ± 85644 (mean ± SD; n=8) and for the TTFields (200 kHz) group was 1,426 ± 1345 (n=6, 2 coverslips had no cell recovery). Average cell diameter was 9.713 ± 0.6312 for the control group and 16.92 ± 2.903 for the TTFields group. TTFields application significantly reduced cell count (P=0.0012) and increased the average cell diameter (P&amp;lt;0.0001) compared to the controls. Conclusions: TTFields application (200 kHz) demonstrated efficacy by markedly reducing cell proliferation (i.e., decreased cell counts) in RCC. The increase in average cell diameter observed with TTFields is consistent with previously reported findings of other research groups also showing increases in cell volume with TTFields in multiple cancer types (brain, lung, pancreas, breast, ovarian, and cervical). This study is the first to demonstrate the efficacy of in vitro TTFields treatment in patient-derived RCC and suggests that patients with renal carcinoma brain metastases may benefit from TTFields therapy. Citation Format: Sharon K. Michelhaugh, Blake C. Walker, Susan Coombe, Sandeep Mittal. Patient-derived metastatic renal carcinoma cells are highly sensitive to in vitro Tumor Treating Fields (TTFields) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1007.