Changes In Cell Counts Research Articles

1265. Effectiveness and Durability of Dolutegravir (DTG)-Based Regimens in Older People Living with HIV (PLWH) from the Veterans Aging Cohort Study (VACS)

Abstract Background HIV management among older people living with HIV (PLWH) may be complicated by the presence of multiple comorbidities and polypharmacy. This study evaluated effectiveness and durability of modern 3-drug antiretroviral regimens among older PLWH. Methods Using data from the Veterans Aging Cohort Study (VACS), PLWH ≥50 years old initiating a dolutegravir (DTG), bictegravir (BIC), elvitegravir (EVG), raltegravir (RAL), or darunavir (DRV)-based 3-drug regimen for the first time between January 1, 2014, and March 31, 2020 were followed from regimen initiation (baseline) until regimen discontinuation (d/c), death, loss to follow-up, or end of study (September 30, 2020). Suppression [viral load (VL)< 50 copies/mL], change in CD4 cell count, and regimen d/c were compared between regimens 6- and 12-months post-baseline using multivariable logistic or linear regression. Virologic failure (VF; 2 consecutive VLs ≥ 200 copies/ml, or 1VL ≥ 200 copies/ml followed by regimen d/c) was evaluated over 12 months. For all outcomes, DTG-based regimens were compared to each other regimen. Outcomes were stratified by treatment experience (ART-naïve and ART-experienced). Results 2,489 ART-naive (DTG: 912, BIC: 432, EVG: 751, RAL: 159, DRV: 235) and 13,810 ART-experienced (DTG: 5097, BIC: 1765, EVG: 3582, RAL: 1486, DRV: 1880) individuals were included. Included PLWH were 97% male and 30% were ≥65 years old (Table 1). For both naive and experienced PLWH, those on DTG were more likely suppressed and had greater increases in CD4 counts at 6 and 12 months compared to those on DRV or RAL (Table 2). Odds of VF did not differ by regimen for ART-naive. For ART-experienced, DTG showed reduced likelihood of VF compared to DRV and RAL. Discontinuations within the first year were higher for RAL and DRV compared to DTG. For ART-experienced PLWH, 6-month d/c was greater for DTG vs. EVG. Regardless of treatment status, no other statistical differences in outcomes were observed between DTG-, BIC-, and EVG-based regimens. Conclusion For both ART-naïve and ART-experienced PLWH >50 years old, treatment responses during the first 12 months of follow-up were similar for those taking DTG-, BIC-, and EVG-based regimens. DTG-based regimens demonstrated greater effectiveness and durability compared to DRV- or RAL-based regimens. Disclosures Cassidy Henegar, PhD, MSPH, GlaxoSmithKline: Stocks/Bonds|ViiV Healthcare: full-time employee Charles Hicks, MD, MD, ViiV Healthcare: I am a full time employee of ViiV Healthcare. Vani Vannappagari, MBBS, MPH, PhD, ViiV Healthcare: I am full time employee of ViiV Healthcare and receive GlaxoSmithKline stock as part of my compensation package|ViiV Healthcare: Stocks/Bonds.

Four Genes Predictive for the Severity of Hematological Damage Reveal a Similar Response after X Irradiation and Chemotherapy.

Radiological and especially nuclear accidents and incidents pose a threat to populations. In such events, gene expression (GE) analysis of a set of 4 genes (FDXR, DDB2, POU2AF1, WNT3) is an emerging approach for early and high-throughput prediction of the later manifesting severity degrees of the hematological acute radiation syndrome (H-ARS). Validation of this gene set on radiation victims is difficult since these events are rare. However, chemotherapy (CTX) is widely used e.g., breast cancer patient treatment and pathomechanisms, as well as blood cell count changes are comparable among both exposure types. We wondered whether GE changes are similarly deregulated after CTX, which would be interpreted as a confirmation of our already identified gene set for H-ARS prediction after irradiation. We examined radiation-induced differential GE (DGE) of our gene set as a positive control using in vitro whole blood samples from ten healthy donors (6 females, 4 males, aged: 24-40 years). Blood was incubated in vitro for 8 h after X irradiation with 0 and 4 Gy (1 Gy/min). These data were compared with DGE measured in vivo in blood samples of 10 breast tumor CTX patients (10 females, aged: 39-71 years) before and 4 days after administration of cyclophosphamide and epirubicin. RNA was isolated, reverse transcribed and quantitative real-time polymerase-chain-reaction (qRT-PCR) was performed to assess DGE of FDXR, DDB2, POU2AF1 and WNT3 relative to the unexposed samples using TaqMan assays. After X irradiation, we found a significant upregulation (irrespective of sex) with mean fold changes of 21 (P < 0.001) and 7 (P < 0.001) for FDXR and DDB2 and a significant down-regulation with mean fold changes of 2.5 (P < 0.001) and 2 (P = 0.005) for POU2AF1 and WNT3, respectively. After CTX, a similar pattern was observed, although mean fold changes of up-regulated FDXR (6-fold, P < 0.001) and DDB2 (3-fold, P < 0.001) as well as down-regulated POU2AF1 (1.2-fold, P = 0.270) and WNT3 (1.3-fold, P = 0.069) appeared lower corresponding to less altered blood cell count changes observed after CTX compared to historic radiation exposure data. However, a subpopulation of CTX patients (n = 6) showed on average a significant downregulation of POU2AF1 (1.8-fold, P = 0.04) and WNT3 (2.1-fold, P = 0.008). In summary, the pattern of up-regulated GE changes observed in all CTX patients and down-regulated GE changes observed in a subgroup of CTX patients appeared comparable with an already identified gene set predictive for the radiation-induced H-ARS. This underlines the significance of in vivo GE measurements in CTX patients, employed as a surrogate model to further validate already identified radiation-induced GE changes predictive for the H-ARS.

Is the definition of late diagnosis correct?

The study analysed 1711 patients of the Hospital for Infectious Diseases in Warsaw diagnosed with HIV infection in 2008-2010 and 2016-2018. Research was conducted examining the changes in CD4 cell counts before starting antiretroviral (ARV) treatment in order to find people who were misclassified as late-diagnosed. Patients with late diagnosis were distinguished on the basis of the consensus definition. The Mann-Whitney U-test was used to analyse the change in CD4 cell counts before starting ARV treatment. In the years 2008-2010, the CD4 count was remeasured before starting ARV treatment in 90 late-diagnosed patients. The median change in the CD4 count was 22 cells/μL. In 49 of these, the number of CD4 cells spontaneously increased before the start of treatment. We can suspect that these patients were misclassified as late-diagnosed. The consensus definition of late diagnosis often leads to overestimation of the number of late-diagnosed patients. The crucial problem is a transient decline in the CD4 lymphocyte count in the acute phase of HIV infection. A potential solution is to introduce serum HIV viral load measurement into the definition.

Airway Inflammation Before and After Bronchial Thermoplasty in Severe Asthma.

Bronchial thermoplasty (BT) is a bronchoscopic treatment for severe asthma, of which the working mechanism and responder profile are partly unknown. The aim of this study is to analyse whether BT alters airway inflammation by epithelial gene expression, inflammatory cell counts and cytokines, and whether this relates to treatment response. In this clinical trial, 28 severe asthma patients underwent bronchoscopy before and after treatment to obtain bronchial brushes and bronchoalveolar lavage fluid (BALF) from treated and untreated airways. RNA was extracted from bronchial brushes for transcriptome analysis, and BALF cells and cytokines were analysed. Asthma quality of life questionnaires were used to distinguish responders from non-responders. We compared results before and after treatment, between treated and untreated airways, and between responders and non-responders. Gene expression of airway epithelium related to airway inflammation gene set was significantly downregulated in treated airways compared to untreated airways, although this did not differ for patients before and after treatment. No differences were observed in cell counts and cytokines, neither from the untreated compared to treated airways, nor before and after treatment. At baseline, compared to non-responders, the expression of genes related to glycolysis in bronchial epithelium was downregulated and both BALF and blood eosinophil counts were higher in responders. Local differences in gene sets pertaining to epithelial inflammatory status were identified between treated and untreated airways after treatment, not resulting in changes in differential cell counts and cytokine analyses in BALF. Secondly, baseline epithelial glycolysis genes and eosinophil counts in BALF and blood were different between responders and non-responders. The observations from this study demonstrate the potential impact of BT on epithelial gene expression related to airway inflammation while also identifying a possible responder profile.

Five-year follow-up of corneal endothelial cell density after transscleral ab interno glaucoma gel stent implantation

PurposeThis study investigates the course of the endothelial cell density over a period of 5 years after XEN45 implantation (XEN45µm, Allergan Plc., USA) with or without combined cataract surgery.MethodsThis is a prospective, cross-sectional, monocentric, non-randomized clinical trial with the intention to treat a population of the University Eye Clinic Glaucoma Service Salzburg. One hundred and fifty-five eyes with preoperative central corneal endothelial cell counts were subjected to XEN45 implantation with (combined surgery group) or without (solo surgery group) combined cataract surgery. Endothelial cell density was measured at 3 corneal positions. XEN45 location parameters were determined with anterior segment OCT and gonioscopy.ResultsIn the combined surgery group, a significant reduction of central endothelial cell count was found at years 2 and 4 when compared to baseline (p = 0.001 and p = 0.02, n = 86), whereas at years 1, 3, and 5, no change was detected (all p > 0.09). The median reduction of endothelial cell count was − 79 (95% CI: − 183 to − 9) and − 93 (95% CI: − 220 to 23) cells at years 2 and 4, respectively. In the solo surgery group (n = 69), no significant change in endothelial cell counts was detected at any time during the 5-year evaluation period (all p > 0.07). Explorative data analyses revealed that XEN45 location parameters did not significantly influence the course of endothelial cell count over time.ConclusionsEndothelial cell loss after XEN45 implantation seems to be low. The present data suggest no impact on the position of the implant with regard to central endothelial cell counts in this study.

Preliminary Study on the Host Response to Bivalent and Monovalent Autogenous Vaccines against Mycoplasma agalactiae in Dairy Sheep.

In Italy, dairy sheep farming represents a vital agro-industry sector, but it is still challenged by contagious agalactia (CA), which is endemic there, and vaccination is the most economical and sustainable tool for control. This study aimed to evaluate the combined Mycoplasma agalactiae (Ma)-Staphylococcus aureus (Sa) vaccine (Ma-Sa) against the Ma monovalent vaccine in ewes. Twelve primiparous Ma-free ewes were randomly grouped into three equal groups: first, the control group injected with placebo, second, the group vaccinated with the Ma monovalent vaccine, and third, the group vaccinated with Ma-Sa combined vaccine, with two S/C doses at 45-day intervals. The animals were examined for serological, hematological, and somatic cell count (SCC) changes for 17 successive weeks. A significant increase in anti-Ma antibody mean titers, leukocytes, and platelets was observed in the vaccinated animals, with the highest values in those who received the combined vaccine. Neutrophils were high only in the animals who received the combined vaccine. SCC was lower in the vaccinated animals during the first six weeks. This study concludes that the combined Ma-Sa vaccines enhance immune response and potentiate its efficacy against Ma. This improvement might be attributed to the sensitization/activation effect of S. aureus on platelets, which are recoded to act as a key regulator for the coordination of all components of the innate immune system. Even though this study included a small number of animals, its findings about the potentialities of this inactivated vaccine in the control of CA are strongly encouraging. Further confirmation might be needed through additional replicates and a challenge study is needed before proceeding with widespread use.

A Bioinformatic Approach to the Clinical Evaluation of Linear Commitment Versus Cloud (Continuum of Low Primed UnDifferentiated) Models for Hematopoiesis Using Plinabulin Clinical Trial Data

Introduction An alternative to the classical Lineage Commitment model for hematopoiesis is the Continuum of Low Primed UnDifferentiated (CLOUD) model (Velten, Nat Cell Biol 2017). Lineage Commitment and CLOUD data are largely derived from in-vitro methods. In-vivo clinical data that is devoid of analytical method-induced 'bias' is sparse. Plinabulin (Plin), is a novel agent that prevented chemotherapy-induced-neutropenia (CIN) in cancer patients (pts) through protection of bone marrow stem cells (Tonra, 2020; Blayney, ASH 2021). Plin, therefore could serve as a pharmacological interventional approach to evaluate Linear Commitment versus CLOUD for hematopoiesis in an in-vivo, human setting. Peripheral blood cell counts (PBCs) from the myeloid, lymphoid, and erythroid lineages in cancer pts receiving myelosuppressive chemotherapy (Chemo) with and without Plin were analyzed using bioinformatics. Method PBCs from cancer pts (Breast, NSCLC, HRPC) receiving Chemo (TAC [docetaxel 75 mg/m2, doxorubicin 50 mg/m2, and cyclophosphamide 500 mg/m2; Study 106; NCT03294577] or docetaxel 75 mg/m2 alone; Study 105 NCT03102606), with Plin doses of 5, 10, 20, 30 mg/m2 (or equivalent) or placebo (Plac; without plinabulin) were collected at screening, predose (cycle (C) 1 day (D) 1,) and at 24 hours after Chemo with or without Plin (C1D2). Plin was given as a single dose by 30 min IV infusion, 30 mins after (the last) Chemo on D1. All pts (with and without Plin) had received dexamethasone 16 mg per day on D-1, 1, and 2 as premedication for docetaxel (and TAC). Blood draws for absolute counts of PBCs, segmented neutrophil (N), band neutrophil (BAND), monocyte (M), eosinophil (E), basophil (B), lymphocyte (L), erythrocyte (RBC), platelet (P), and immature cells, were analysed by Central Laboratory (Covance). Bioinformatic analysis was performed by FIOS Genomics. To detect treatment-specific effects, exploratory principal component analysis (PCA) and Pearson correlation analysis were performed. 'Limma' linear modelling was used to assess treatment impact on cell counts (C1D1 vs screening; C1D2 vs C1D1) and changes in cell counts (screening to C1D1; C1D1 to C1D2), adjusting for relevant clinical variables. Further correlation and clustering analyses per hematopoietic hierarchy (MEP, GMP, and GLP) were performed to determine correlations between change in cell abundance on C1D2 vs C1D1 for Plin and Plac. Significant differences were defined as those with a P-value adjusted for multiple testing of P < 0.05. Result Following Plac, cell types from myeloid, lymphoid, and erythroid lineage demonstrated significantly decreased abundance on C1D2 vs C1D1, reflecting myelosuppression by Chemo. In contrast, following Plin, an increase in abundance for all GMP-derived myeloid cells (N, M, B, and E) were observed, and in Plin dose-dependent manner. In absolute counts, GMP-derived myeloid cells had increased significantly with Plin at C1D2 vs C1D1, suggesting not only protection, but also stimulation of GMP cells by Plin. Furthermore, significant increases on C1D2 vs C1D1 in immature post-GMP progenitor cells (BAND, metamyelocytes, and myelocytes), were observed with Plin, but not with Plac, further suggesting GMP stimulation by Plin. For CLP and MEP-derived cells, decreases were seen with Plin, however, to a much smaller extent compared to Plac, suggesting some degree of protection with Plin. Cell cluster analysis showed inter-correlation in abundance of GMP-derived cell types (N, M, B, and E), which is in support Lineage Commitment. Relative abundance in cells derived from the CLP and MEP were also intercorrelated with abundance of GMP-derived cells, further supporting Lineage Commitment. Conclusion Using bioinformatic analyses, we demonstrated that Plin not only exerts protective, but also stimulatory effects on GMP cells under myelosuppressive conditions. The abundance of immature PBCs derived from the GMPs is in further support of stimulation of GMP cells with Plin. The data also suggests a degree of protective effect with Plin on CLP- and MEP-cells. The interdependence in abundance of mature PBCs derived from GMP cells, but also from CLP- and MEP-derived cells, is strongly supportive of Lineage Commitment. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Rapid Tumor Debulking of Relapsed/Refractory CLL Patients By PI3Kδ Inhibition and Anti-CD20 Monoclonal Antibody Treatment

Introduction Effective multi-targeted drug combinations could rapidly achieve deep remissions in CLL allowing for defined duration treatment. The combination of a novel highly-specific phosphoinositide 3-kinase delta (PI3Kδ) inhibitor and casein kinase 1 epsilon (CSK1ε) inhibitor, umbralisib (UMB), and a glycoengineered chimeric monoclonal antibody (mAb) targeting a unique epitope on CD20, ublituximab (UBL), with the BCL2 inhibitor venetoclax (VEN) may decrease drug resistance, reduce risk of tumor lysis syndrome (TLS) and achieve higher levels of undetectable minimal residual disease (MRD). This triple agent therapy is being evaluated in relapsed or refractory CLL patients (U2-VEN, ClinicalTrials.gov NCT03379051). Patients received oral 800 mg UMB once-daily. For Cycle 1, UBL was given via IV on Day 1 (150 mg), Day 2 (750 mg), Day 8 (900 mg), Day 15 (900 mg), and then on Day 1 in Cycles 2 and 3 (900 mg); for 3 cycles of 28 days, followed by consolidation with UMB and VEN for cycles 4 to 24. Patient (n = 25) blood samples were collected Pre- and Post-treatment on days of UBL administration during cycles 1-3. Our studies focused on UMB and UBL treatment effects on: 1) The decrease in the circulating CLL tumor burden and thus the subsequent risk of TLS from the addition of VEN to the regimen; and 2) CD20 antigen loss on CLL cells, a major mechanism of anti-CD20 mAb resistance. Methods Blood was analyzed by clinical laboratory automated cell counter and then stained with antibodies against CD3, CD5, CD19, and CD20 for flow cytometry analysis. CLL cell counts were calculated from flow cytometry and clinical laboratory results. To quantitate CD20 surface molecules, peripheral blood mononuclear cells were isolated and stained for flow cytometry analysis with the addition of a Molecules of Equivalent Soluble Fluorochrome standard curve. All human specimen collection and usage was conducted with written informed consent after approval of our Institutional Research Subjects Review Board according to the ethical guidelines of the Declaration of Helsinki. Results Average CLL cell counts (47 x 109/L) decreased rapidly after administration of the first dose of UMB and UBL (C1D1 Post, 26 x 109/L), which rebounded the next day (C1D2 Pre, 43 x 109/L) and then decreased after the second treatment (C1D2 Post, 24 x 109/L). By Day 15, the average CLL cell count had dropped to 9 x 109/L, which decreased further to 5 x 109/L by the start of cycle 3 (Fig. 1). Notably, the variation in CLL cell counts (error bars) decreases over treatment time, suggesting that most CLL patients' tumor burdens are decreasing to a similar low level independent of starting CLL cell count. The change in average CLL cell count between Pre and Post measurements is initially very high (45% decrease C1D1) with decreasing effectiveness over treatment time (8% decrease C3D1). This change could be due to the loss of CLL cell membrane CD20 levels leading to mAb resistance. Pre-treatment (C1D1 Pre) average CLL cell membrane CD20 levels (n=18) were 29,000 CD20 molecules/cell. This decreased to 9,000 molecules/cell after the first treatment (C1D1 Post), stabilized at 9,100 molecules/cell on the next day (C1D2 Pre) and then decreased further after the second treatment (C1D2 Post) to 1600 molecules/cell. After day 8 treatment, CD20 levels remained at 1,000 molecules/cell or less until the end of this study (C3D1). Discussion UMB and UBL rapidly decrease circulating CLL counts over three cycles of treatment, reducing most CLL patient tumor burden to similar low levels. This clinical response could facilitate addition of VEN in the 4thcycle of therapy by decreasing the risk of TLS. The rapid loss of surface CD20 from CLL cells to 1,000 CD20 molecules/cell or less by C1D8 Post has the potential to cause acquired resistance to anti-CD20 mAb therapy. This rapid and sustained loss of CD20 levels is consistent with that reported at standard doses for other anti-CD20 mAbs. However, an effect of concomitant UMB on CD20 levels cannot be ruled out. Previous work suggests that lower anti-CD20 mAb doses can prevent these large decreases in CD20 levels and increase UBL efficacy. In conclusion, our initial data show that treatment with a selective PI3Kδ and CSK1ε inhibitor (UMB) combined with an anti-CD20 mAb (UBL) is effective in decreasing CLL cell counts in all patients assessed in the U2-VEN clinical trial. However, treatment efficacy could be limited by large decreases in the CLL surface CD20 levels. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Prognostic Implication of Exfoliative Airway Pathology in Cancer-Free Coal Workers' Pneumoconiosis.

The purpose of this study is to see if exfoliative pulmonary airway pathology in cancer-free coal workers' pneumoconiosis (CWP) can be used as a biomarker for predicting pulmonary morbidity. We investigated persistent metaplastic changes in bronchoscopic washing cytology and differential cell counts in bronchoalveolar lavages (BAL) in 97 miners with CWP and 80 miners without CWP as the control. Clinicopathological parameters were examined including pulmonary function tests and the presence of progressive massive fibrosis. When compared to the control group, severe alveolitis, severe goblet cell hyperplasia (GCH), severe hyperplastic epithelial change, and severe squamous metaplasia were the distinguishing biomarkers in CWP. Multivariate analysis revealed that severe alveolitis and severe GCH, along with miner duration and current smoker, were independent predictors of pulmonary mortality. The survival analysis revealed a significantly different survival rate between the three groups: no evidence of severe alveolitis and severe GCH, presence of severe alveolitis or severe GCH but not both, and both severe alveolitis and severe GCH. The severities of alveolitis and goblet cell hyperplasia in the bronchoscopic study are independent prognostic factors for CWP. A pathologic grading system based on these two parameters could be used in the stratification and clinical management of CWP patients.

HSC Niche Dynamics in Regeneration, Pre-malignancy, and Cancer: Insights From Mathematical Modeling.

The hematopoietic stem cell (HSC) niche is a crucial driver of regeneration and malignancy. Its interaction with hematopoietic and malignant stem cells is highly complex and direct experimental observations are challenging. We here develop a mathematical model which helps relate processes in the niche to measurable changes of stem and non-stem cell counts. HSC attached to the niche are assumed to be quiescent. After detachment HSC become activated and divide or differentiate. To maintain their stemness, the progeny originating from division must reattach to the niche. We use mouse data from literature to parametrize the model. By combining mathematical analysis and computer simulations, we systematically investigate the impact of stem cell proliferation, differentiation, niche attachment, and detachment on clinically relevant scenarios. These include bone marrow transplantation, clonal competition, and eradication of malignant cells. According to our model, sampling of blood or bulk marrow provides only limited information about cellular interactions in the niche and the clonal composition of the stem cell population. Furthermore, we investigate how interference with processes in the stem cell niche could help to increase the effect of low-dose chemotherapy or to improve the homing of genetically engineered cells.

Functional MRI radiomics-based assessment of pelvic bone marrow changes after concurrent chemoradiotherapy for cervical cancer

ObjectivesTo quantify the dose-response relationship of changes in pelvic bone marrow (PBM) functional MR radiomic features (RF) during concurrent chemoradiotherapy (CCRT) for patients with cervical cancer and establish the correlation with hematologic toxicity to provide a basis for PBM sparing.MethodsA total of 54 cervical cancer patients who received CCRT were studied retrospectively. Patients underwent MRI IDEAL IQ and T2 fat suppression (T2fs) scanning pre- and post-CCRT. The PBM RFs were extracted from each region of interest at dose gradients of 5–10 Gy, 10–15 Gy, 15–20 Gy, 20–30 Gy, 30–40 Gy, 40–50 Gy, and > 50 Gy, and changes in peripheral blood cell (PBC) counts during radiotherapy were assessed. The dose-response relationship of RF changes and their correlation with PBC changes were investigated.ResultsWhite blood cell, neutrophils (ANC) and lymphocyte counts during treatment were decreased by 49.4%, 41.4%, and 76.3%, respectively. Most firstorder features exhibited a significant dose-response relationship, particularly FatFrac IDEAL IQ, which had a maximum dose-response curve slope of 10.09, and WATER IDEAL IQ had a slope of − 7.93. The firstorder-Range in FAT IDEAL IQ and firstorder-10Percentile in T2fs, showed a significant correlation between the changes in ANC counts under the low dose gradient of 5–10 Gy (r = 0.744, -0.654, respectively, p < 0.05).ConclusionFunctional MR radiomics can detect microscopic changes in PBM at various dose gradients and provide an objective reference for bone marrow sparing and dose limitation in cervical cancer CCRT.

A case of natalizumab‐associated progressive multifocal leukoencephalopathy followed by immune reconstitution inflammatory syndrome with difficulty in the timing of immunotherapy

AbstractBackgroundDetails regarding the clinical course of natalizumab‐associated progressive multifocal leukoencephalopathy (NAT‐PML) have not been reported in Japanese patients. We experienced a Japanese NAT‐PML case and report it for the purpose of clarifying the challenge it posed in medical treatment.Case PresentationHerein, we describe a 58‐y‐old multiple sclerosis patient who had NAT‐PML with immune reconstitution inflammatory syndrome (IRIS). Before NAT‐PML developed, the patient's Expanded Disability Status Scale (EDSS) score was 5.5. She received natalizumab (300 mg) every 3–7 w, and her EDSS score had not changed for 9.1 y. The John Cunningham virus (JCV) index was 0.43 before NAT‐PML developed. She developed a gait disturbance when NAT‐PML emerged. Natalizumab was administered a total of 108 times. The patient was diagnosed with probable NAT‐PML on the basis of punctate lesions on T2‐weighted imaging and a JCV‐PCR result of 29 copies/ml from cerebrospinal fluid (CSF) testing. Intravenous methylprednisolone (IVMP) was initiated when the contrast‐enhanced lesion was first detected, but the NAT‐PML lesion was rather enlarged despite the temporary disappearance of the contrast‐enhanced lesion. An obvious increase in the IgG index and a slight increase in the CSF cell count were recognized at the time the immunological response was activated. Three cycles of a 3‐d course of IVMP were administered every 2 w for IRIS, and the patient's worst EDSS score was 9.5.ConclusionTreatment sequencing should be executed before the onset of NAT‐PML. Changes in CSF cell count and IgG index may be useful for treatment decision; further research is needed.

Differential diagnosis of leukocytosis and leukopenia.

The blood cell count is often examined in routine clinical praxis. Physiologic leucocyte count is in range 4-10 × 109 in liter of blood. Abnormal values of leukocytes and subtypes of leukocytes in differential count are often present. Changes in leukocytes counts are caused by variety of benignant or malignant conditions. It is important in clinical praxis to interpret changes in blood cell count correctly and choose adequate approach in investigation process. In general, leukocytosis and leukocytopenia may present in primary hematologic disorder or secondary/reactive states, caused by reaction of hematopoiesis to underlying condition. This article review common causes of leukocytosis or leucopenia and give basic advice how to investigate patients with changes in leukocytes count.

Effects of Combined Visible and Infrared Light Rhinophototherapy in Patients With Allergic Rhinitis.

Intranasal phototherapy offers an alternative treatment method for patients with allergic rhinitis who cannot benefit from intranasal corticosteroids and oral antihistamines. Different wavelengths have been tried with promising results. In this present study, we aimed to investigate the effects of visible light-infrared light phototherapy on clinical improvements together with its cytologic effects in patients with allergic rhinitis. Patients with confirmed allergic rhinitis were given a 4-week course of intranasal phototherapy treatment. Weekly symptom questionnaires were applied to monitor clinical effects. Nasal lavage specimens were obtained before the start and at the completion of the 4-week therapy. Fluorescence-activated cell sorting analyses of CD16+, CD24+, and CD 45+ cells were performed. Statistical analyses are performed of weekly changes in symptoms and cell counts. CD45+CD16highCD24+ neutrophil count in nasal lavages decreased significantly whereas CD45+CD16dim/-CD24+ eosinophil counts significantly increased and CD45+ granulocyte counts remained unchanged. Symptom scores including nasal itching, nasal discharge, nasal obstruction, sneezing, eye itching, throat itching, and ear itching all statistically decreased compared to baseline at the end of 4 weeks. Four-week course of intranasal phototherapy with visible and infrared light leads to clinical improvement in allergic rhinitis patients.

State of refractive surgery with Visian ICL posterior chamber phakic lens in Spain

ObjectivesTo analyse the state of refractive surgery with Visian ICL phakic lens implantation in Spain, indicating the mean ranges of implanted powers, visual and refractive outcomes obtained and safety and efficacy indices. MethodThis is a population-based, cross-sectional, descriptive, analytical, retrospective, multicentre study in which each co-author collected data from at least the last 10 patients (up to a maximum of 30) with Visian ICL phakic lens implantation. One eye from each subject was randomly selected for study inclusion, and the variables analysed were: age, gender, visual acuities with and without correction pre and post-surgery, preoperative and residualrefraction, keratometry, endothelial cellular density, anterior chamber depth from endothelium, central corneal pachymetry, intraocular pressure, white-white, implanted lens (power and size), expected and achieved spherical equivalent (SEQ) post-surgery at one month after surgery. ResultsA sample of 140 eyes was obtained from 140 patients with a mean age of 31.35 ± 7.28 years. Mean preoperative SEQ was −6.33 ± 3.69D, mean CDVA was 0.96 ± 0.16, ACD was 3.30 ± 0.29 mm, WTW 12.02 ± 0.40 mm, CCT 535.03 ± 37.68 µm, ECD 2684.37 ± 313.74 cels/mm2 and IOP 14.84 ± 2.59 mmHg. 66.4% of the implanted lenses were spherical ICLs and 33.6% toric, with a mean power of −7.81 ± 4.09D and a mean cylinder of 2.27 ± 1.23D. 5.7% of the implanted lenses were hypermetropic. The mean SEQ target selected was +0.04 ± 0.27D. 48.92% of the implanted lenses were of size 13.2 mm, with the remaining 36.69, 10.79 and 3.60% being of sizes 12.6, 12.1 and 13.7 mm, respectively. At one month after surgery a residual SEQ of −0.01 ± 0.31D was obtained. The efficacy and safety indices were 1.06 ± 0.18 and 1.10 ± 0.19, respectively. The mean central vault was 508.80 ± 201.04 µm. The accuracy between the calculated SEQ and the obtained SEQ was 99.3% for spherical lenses and 88.2% for Cyl correction. The mean ECD at 1 month after surgery was 2805.53±273.30céls/mm2, which was a difference from preoperative of 0.11% (p = 0.922), similarly, the mean IOP was 13.98 ± 2.57 mmHg, which was a difference of −0.92 ± 2.80 mmHg. ConclusionsICL phakic lens surgery is an effective, safe and predictable procedure. There were no significant changes in endothelial cell count or intraocular pressure measurement one month after surgery.

Stereological examination of the effect on the hippocampal dentate gyrus granule cells number of rats subjected to wireless internet during prenatal period: a preliminary study

As a result of industrialisation and the many recent advances in technology, we are being intensely subjected to non-ionised radiation sources. As such, the effect of non-ionised radiation on human tissue is now being researched. In this study, the effects of Wi-Fi waves found in some third-generation mobile phones to facilitate internet connection on the brain hippocampal dentate gyrus granule cell quantity of rats during the prenatal period was stereologically examined. During the study, mated albino rats were subjected to Wi-Fi throughout pregnancy. A month after giving birth, six rats from each group, for a total of 12 rats, was sacrificed under perfusion and anaesthesia. Their skulls were opened and brains were removed to undergo routine checks in which the granule cells were counted. Strategic sections were defined, and every 45th section couple (with a thickness of 5 μm) was taken and dyed with haematoxylin and eosin (H&amp;E) and cresyl violet stain. The granule cells were counted using a combination of the stereological disector method and Cavalieri’s principle. Then, the results were analysed using the Mann–Whitney U test. No statistically significant difference has been observed between the groups (p &gt; 0.05). The findings were discussed in light of the relevant literature, and it was determined that throughout pregnancy, Wi-Fi modem device did not result in any changes in the dentate gyrus granule cell count of the hippocampus of postnatal rats’ brains.

Serum cytokine levels are associated with tumor progression during FOLFIRINOX chemotherapy and overall survival in pancreatic cancer patients.

BackgroundBiomarkers predicting treatment response may be used to stratify patients with pancreatic ductal adenocarcinoma (PDAC) for available therapies. The aim of this study was to evaluate the association of circulating cytokines with FOLFIRINOX response and with overall survival (OS).MethodsSerum samples were collected before start and after the first cycle of FOLFIRINOX from patients with PDAC (n=83) of all disease stages. Overall, 34 circulating cytokines were analyzed with a multiplex immunoassay. In addition, changes in peripheral blood immune cell counts were determined by flow cytometry to correlate with differences in cytokine levels. Chemotherapy response was determined by CT scans with the RECIST 1.1 criteria, as disease control (n=64) or progressive disease (n=19) within eight cycles of FOLFIRINOX.ResultsPatients with high serum IL-1RA concentrations after one cycle of chemotherapy were less likely to have tumor progression during FOLFIRINOX (OR 0.25, P=0.040). Increase of circulating IL-1RA concentrations correlated with increase of total, classical (CD14+CD16-), and non-classical monocytes (CD14-CD16+), and dendritic cells. In multivariable cox regression, including the variables chemotherapy response outcome and baseline CA19-9 level, serum concentrations of IL-7 (HR 2.14, P=0.010), IL-18 (HR 2.00, P=0.020), and MIP-1β (HR 0.51, P=0.025) after one cycle of FOLFIRINOX showed correlations with OS.ConclusionsCirculating IL-1RA, IL-7, IL-18, and MIP-1β concentrations are biomarkers associated with FOLFIRINOX response in PDAC patients, suggesting an important role for specific immune cells in chemotherapy response and PDAC progression. Cytokine-based treatment might improve patient outcome and should be evaluated in future studies.

MDSCs Aggravate the Asthmatic Progression in Children and OVA-Allergic Mice by Regulating the Th1/Th2/Th17 Responses.

Background Asthma is a chronic inflammatory disease of respiratory with serious risks for children. This study explored myeloid-derived suppressor cells (MDSCs) on the pathogenesis of asthmatic children and mice. Methods The clinical study enrolled 30 asthma, 20 pneumonia, and 20 control participants. The MDSCs, Th17 and Th1 cells percentage, and IL-4, IL-12, IL-10, and IFN-γ levels were detected by flow cytometry and ELISA. In experimental asthma, mice were divided into control, ovalbumin (OVA), and OVA + MDSCs groups. The changes in inflammatory cell count and the levels of IL-5, IL-12, and IL-10 in mice BALF and the levels of inflammatory factors, IgE, and IFN-γ in mice were detected by ELISA. The amount of ROS generation and pathological changes and the levels of caspase 1 and caspase 3 were tested by flow cytometry, HE and PAS staining, and immunohistochemistry. The expression of cleaved caspase 1/caspase 1 and cleaved caspase 3/caspase 3 was detected by western blot. Results In clinical trials, the levels of IL-12, IFN-γ, and Th1 percentage decreased in pneumonia and asthma children's peripheral blood, while the levels of IL-4 and IL-10 and the percentages MDSCs and Th17 increased. In asthma mice, pathological staining showed that asthma caused lung inflammation and damage, while the OVA + MDSC group was severer. Moreover, the percentages of eosinophils, neutrophils, lymphocytes, and the levels of inflammatory factors, IgE, ROS production, caspase 1, caspase 3, cleaved caspase 1/caspase 1, and cleaved caspase 3/caspase 3 increased in OVA + MDSC group, while the percentage of macrophages, IL-12, and IFN-γ levels reduced, illustrating that MDSCs exacerbated asthma. Conclusion Our study indicated that MDSCs could aggravate asthma by regulating the Th1/Th2/Th17 response.

Platelet-derived immuno-inflammatory indices show best performance in early prediction of COVID-19 progression.

BackgroundCoronavirus disease 2019 (COVID‐19) profoundly affects the immune and hematopoietic systems with various degrees of reactive changes in the blood cell counts. Immuno‐inflammatory indices are considered a simple and effective tool in the prediction of COVID‐19 outcomes. We aimed to evaluate and compare the usefulness of leukocyte and platelet counts‐based immuno‐inflammatory indices on admission to hospital in predicting COVID‐19 progression and mortality.MethodsA total of 945 patients were enrolled. In addition to blood cell counts, we assessed hemogram‐derived immuno‐inflammatory indices in relation to COVID‐19 progression and death. The indices were tested by analysis of variance, receiver operating characteristic curve analysis, and binomial logistic regressions.ResultsPatients with severe COVID‐19 had significantly higher counts of neutrophils, eosinophils, and large immature cells (LIC), while decreased counts of platelets and monocytes. Lymphopenia was found in all of the patients, but without significant association with the outcomes. Patients with a LIC count ≥0.265 x 09/L had 54.7% more odds of having COVID‐19 progression. In multivariable analyses, platelets/neutrophil‐to‐lymphocyte ratio (P/NLR) and platelets‐to‐neutrophil radio (P/N) were significant independent predictors of COVID‐19 progression and mortality. The odds of a poor outcome were two times higher in cases with P/NLR < 43 x 109/L and P/N < 29 x 109/L.ConclusionIndices that include platelet count in combination with neutrophil and/or lymphocyte counts displayed the best discriminatory ability and prognostic value of COVID‐19 outcomes. Additionally, LIC showed promising results in the early identification of severe COVID‐19.

White blood cell count: a valuable tool for suspecting Cushing's syndrome.

Simple screening tests to determine whether Cushing's syndrome (CS) should be ruled out are lacking. Tools that enable early diagnosis could reduce morbidity and associated sequelae. The potential of glucocorticoid-induced changes in the white blood cell (WBC) count for raising suspicion of CS is assessed. This was a retrospective case‒control study. The WBC counts of 73 cases with CS and 146 matched controls were compared. The number of leukocytes (Leu), the number and percentage of neutrophils (N, Np), the number and percentage of lymphocytes (L, Lp), neutrophil-to-lymphocyte differences in the number and percentage (N-L, Np-Lp), neutrophil-to-lymphocyte ratio in the number and percentage (NLR, NLRp), and leukocyte-to-lymphocyte differences (Leu-L) were evaluated. The area under the ROC curve (AUC) was calculated for each of these parameters. Reference values were estimated that could help disclose occult CS. All ten parameters showed significant differences between cases and controls. The AUC was greater than 0.7 for all ten parameters, and was the best for the NLRp and Lp (AUC: 0.89). An Lp of 23.9% showed a diagnostic accuracy of 84.9% for the diagnosis of CS. The concordance of an Lp below 24% and more than 8000 leucocytes had a PPV of 78.2% for CS, while the pairing of an Lp over 24% and a Leu below 8000 cells had an NPV of 97.3% for CS. WBC count assessment can be an effective tool to raise suspicion of CS, prompting diagnostic testing. This simple and universally available test may allow earlier diagnosis of CS before highly evolved phenotypes develop.