Becton Dickinson Research Articles

Comprehensive assessment of vaginal infections using a single swab

BackgroundThe decision to use a particular test to diagnose patients presenting with symptoms of vaginitis and/or STI is based primarily on the prevailing standards of care in the clinic at...

A-097 Stability Study of 21 Common Plasma Biochemistries in Unspun Whole Blood

Abstract Background Most blood collection tube manufacturers typically recommend separating serum/plasma from red blood cells within 2 h of collection prior to biochemical analysis to minimize cell contact and thus maintain stability of analytes. This can lead to rejected testing when uncentrifuged samples are transported from ambulatory sites to core laboratories over long distances, leading to delayed patient care and decreased patient satisfaction from having to be redrawn. Few studies are available to make recommendations of specimen acceptance with delayed centrifugation time. Our study aimed to assess whole blood stability of routinely ordered chemistry tests. Methods Between November 30–December 1, 2022, venous blood from 20 non-fasting healthy adults(30% male) with ages ranging from 23 to 57 years were collected into 3-mL plastic Becton Dickinson(BD) Vacutainer™plasma separated tubes(PST). For each person, 6 PST were collected. One whole blood sample was centrifuged immediately(T0) at 4000g for 3 min at room temperature(RT) and analyzed. Remaining 5 whole blood samples were stored at either RT or 4 °C and the subsequent centrifugation of the samples occurred at 2, 4, 6, 12 and 24 h after collection. The analytes studied using Abbott Alinity instrumentation were comprehensive metabolic panel(CMP), lipid panel, TSH and FT4. Absolute and percent bias at various time points for each participant were calculated using the reference sample(T0). Observed biases were compared to allowable total error based on desirable biological variation(BV) from European-Federation-of-Laboratory-Medicine(EFLM) database and Clinical-Laboratory-Improvement-Amendments(CLIA) 2024 upcoming limits. Results Analytes stable in the whole blood at both RT and 4 °C up to 24 h based on EFLM-BV and CLIA were ALP, ALT, AST, BUN, cholesterol, creatinine, HDL, total bilirubin, triglycerides, and TSH. Analytes with variable stability times are shown in the table below: Conclusion This study provides a guideline of acceptable times for whole blood samples sent for routine chemistry tests when centrifugation was not performed within 2 h from collection.

Accuracy, Precision, and Residual Volume of Commonly Used Syringes for Intravitreal Injections and the Impact on Intraocular Pressure

To compare accuracy, precision, and residual volume of commonly used syringes for intravitreal injections (IVIs) and to assess the intraocular pressure (IOP) rise by variations in volumes delivered. Experimental laboratory study. No subjects were involved in this study. We tested 8 syringe models with 2 different needle setups, with 2 different solutions (distilled water or glycerin) and target volumes (50 and 70 μL). To obtain the delivered and residual volumes, we weighed the syringe-needle setups with scale before liquid withdrawal, with liquid, and after liquid release. We also created an experimental eye model to determine the transient rise in IOP following stepwise 10-μL increases in injection volumes. Delivered and residual volumes, IOP rise. We tested a total of 600 syringe-needle setups. Becton Dickinson (BD) Ultra-Fine (0.34 ± 0.28 μL), Zero Residual (1.53 ± 1.15 μL), and Zero Residual Silicone Oil-free (1.40 ± 1.16 μL) syringes showed the lowest residual volume (P < 0.001) in comparison with the others (range: 24.86 ± 1.78 μL for Injekt-F to 51.97 ± 3.37 μL for Omnifix-F). The most accurate setups were (percentage deviation from target volume): Zero Residual Silicone Oil-free (+ 0.70%), Zero Residual 0.3 ml (+ 4.49%), BD Ultra-Fine (+ 7.83%), Injekt-F (9.42%), Norm-Ject (+ 15.88%), Omnifix-F (+ 16.96%), BD Plastipak Brazil (+17.96%), and BD Plastipak Spain syringes (+ 19.41%). There was a statistically significant difference between the Zero Residual Silicone Oil-free syringe and all other syringes (P < 0.0001), except for the Zero Residual 0.3-ml syringe (P= 0.029). The coefficient of variation was low for all syringes. The modeled IOP rise ranged from 32.3 (standard deviation [SD], 1.4) mmHg for 20-μL injection volume to 76.5 (SD, 1.0) mmHg for 80-μL injection volume. For the standard 50-μL injection volume, the peak pressure was 50.7 (SD, 0.1) mmHg, and the pressure rise duration was 28 (SD, 2) minutes. There were significant differences in accuracy and residual volume between syringes, whereas they showed a high precision. Volume excess results in a considerable increase in IOP rise after injection. These findings may provide a relevant overview to clinicians and to both device and drug manufacturers regarding pharmacoeconomic, safety, and efficacy issues. Proprietary or commercial disclosure may be found after the references.

Direct inoculation method for identification and antimicrobial susceptibility testing using matrix-assisted laser desorption ionization-time of flight mass spectrometry and both the Vitek 2 and MicroScan Walkaway 96 Plus systems

The aim of our study was to evaluate a protocol utilizing serum separator tubes (SST) to facilitate a faster, cost-effective, direct method for rapid sensitivity testing and identification of positive blood cultures. Spiked cultures were inoculated into either Becton Dickinson (BD) BACTECTM Aerobic Plus or Anaerobic/F bottles containing sterile human blood. Bottles were immediately processed when positive. A parallel study using patient isolates was used in which bacteria were pelleted by SST from positive blood cultures. For identification, a portion of the pellet was tested by matrix-assisted laser desorption/ionization as described by the manufacturer. MicroScan panels and Vitek 2 results were compared. Categorical agreement was used as comparison to standard subculture and/or polymerase chain reaction methods. No discordant identifications were observed, and 86% generated a successful identification when compared to subculture methods. For the Vitek 2, we observed a 99% essential agreement when compared to the subculture method. For the MicroScan Walkaway, we observed 94.9%, 97.4%, and 100% categorical agreement for MIC panels 53, 38, and MICroSTREP Plus 2, respectively. Turnaround times were reduced from 4 hours for identification and 11 hours for antimicrobial sensitivity testing. We conclude that the SST method results in timelier, actionable results for antimicrobial stewardship initiatives.

Expanding comprehensive medication management considerations to include responses to the social determinants of health within the BDHelping Build Healthy Communities Program

Expanding comprehensive medication management considerations to include responses to the social determinants of health within the <scp>BD</scp><i>Helping Build Healthy Communities Program</i>

Frequency and Symptoms of Intravitreal Silicone Oil Droplets Following Becton Dickinson and Norm-Ject Bevacizumab Injections.

To determine silicone oil droplet frequency and symptomatic impact in patients injected with Norm-Ject (NJ) and/or Becton Dickinson (BD) intravitreal bevacizumab. This was a retrospective cohort study of 426 patients with prior bevacizumab injection(s). Symptomatic floaters questionnaire responses were compiled and statistical analysis was performed using Fisher's exact t test with 95% CI calculated via the modified Wald method. Patients who received BD intravitreal bevacizumab showed more droplets (67.2%) than those who received NJ intravitreal bevacizumab (7.8%), and droplets increased with injection quantity. However, the symptomatic patients reporting new floaters were similar (NJ: 39.22%, BD: 39.47%). [Ophthalmic Surg Lasers Imaging Retina. 2022;53:108-112.].

DETECTION OF COLISTIN RESISTANCE IN MULTIDRUG-RESISTANT GRAM-NEGATIVE BACILLI: PERFORMANCE OF THE BD PHOENIX AUTOMATED SYSTEM VERSUS THE BROTH MICRODILUTION METHOD

Introduction: The increased dissemination of multidrug-resistant Gram-negative bacilli along with a lag in the development of novel, broad-spectrum antimicrobials determined the reintroduction of colistin into clinical practice. In this context, antibiotic susceptibility testing for colistin using high-performance methods has become a requirement. Objective: We compared the performances of the Becton Dickinson (BD) Phoenix 50 automated system and the Micronaut MIC-Strip, a commercial broth microdilution (BMD) method, for the detection of colistin resistance in clinical multidrug-resistant Gram-negative bacilli, isolated from patients admitted in a tertiary hospital in southern Romania. Methods: 54 clinical isolates with different multidrug-resistant phenotypes (Enterobacterales and Pseudomonas aeruginosa) were tested for colistin susceptibility with the BD Phoenix 50 automated system and the Micronaut MIC-Strip, following the Clinical Laboratory Standard Institute recommendations (CLSI, 2021) and were further classified according to the interpretative categories (resistant and intermediate, respectively). The statistical data was calculated using Microsoft Excel 2007 and OpenEPI software. Results: BD Phoenix system failed to detect 11 colistin resistant isolates (false susceptibility). By contrast, all 38 clinical isolates defined as intermediate by the BMD method were also classified as intermediate by the Phoenix system. Conclusion: BD Phoenix system is a reliable technology for detecting colistin resistance. However, high rates of false susceptibility were observed for the BD Phoenix system, indicating that the intermediate / susceptible results should be confirmed with the standard BMD method. Keywords: colistin susceptibility testing, Gram-negative bacilli, multidrug resistance.

Occurrence of Multiple, Extensive and Pan Drug-Resistant Pseudomonas aeruginosa and Carbapenemase Production from Presumptive Isolates Stored in a Biobank at Ethiopian Public Health Institute.

PurposePseudomonas aeruginosa is a common cause of nosocomial infections with associated morbidity and mortality because the organism is unresponsive to commonly available antimicrobials. This study was undertaken to determine the multiple drug-resistant (MDR), extensive drug-resistant (XDR) and pan drug-resistant (PDR) phenotype of P. aeruginosa and its carbapenemase production rate from presumptive isolates stored in the biobank at the Ethiopian Public Health Institute (EPHI).MethodsA cross-sectional study was conducted at the EPHI laboratory, Addis Ababa, Ethiopia from March to June 2021. Stored isolates of Pseudomonas spp. which had been characterized by manual identification methods were further processed for species-level identification (ID) and antimicrobial susceptibility testing (AST) using a Becton Dickinson (BD) Phoenix automated system. The isolates were analyzed for carbapenemase enzyme production using the modified Carbapenem Inactivation Method (mCIM). The data analysis was done using SPSS version 20 software.ResultsIn this study, 110 presumptive Pseudomonas isolates from a biobank were re-analyzed, 100 of them were found to be Pseudomonas and among these P. aeruginosa accounted for 98% and P. putida accounted for 2%. The majority of isolates were recovered from wound (46%) specimens followed by ear swabs (18%). The highest level of resistance was observed against ceftazidime (35%) and the lowest level of resistance was observed against amikacin (2%). Twenty-seven isolates were identified as candidates for carbapenemase enzyme production testing, of which only 3/27 (11%) isolates were detected as carbapenemase enzyme producers.ConclusionThis study shows an increasing rate of MDR and XDR isolates and the appearance of PDR in P. aeruginosa strains; this is a serious problem in Ethiopia. The lack of newer anti-pseudomonal antibiotics adds to the problem. In order to alleviate this, infection prevention activities should be promoted, and treatment of bacterial infections should be guided by antibiotic susceptibility test results.

Artemisinins Synergize with BCL2 Inhibitors By Reducing MCL1 Protein Levels in Leukemia Cells

Artemisinins have broad antineoplastic activity in vitro , are well tolerated and inexpensive, and can be parenterally or orally administered in humans. Both artesunate (AS; the current World Health Organization-recommended drug for severe malaria) and artemisinin-derived trioxane diphenylphosphate dimer 838 (ART838; a more potent derivative) substantially inhibited acute leukemia growth in vivo and in vitro , at doses where normal CD34+ hematopoietic stem-progenitor cell clonogenicity was essentially unaffected (Fox et al, Oncotarget 2015, PMID: 26771236).To investigate integration of artemisinins into the treatment of acute leukemias, we sought to determine, via a focused drug combination screen, if AS and ART838 could be paired synergistically with emerging antileukemic drugs. Navitoclax (ABT737) and venetoclax (ABT199), the only BCL2 inhibitors in the drug combination screen library of 111 emerging antineoplastic compounds, were 2 of the top 3 candidates identified, synergizing with both AS and ART838 against all 3 acute leukemia cell lines screened. Independent in vitro and in vivo experiments validated the screening results that AS and ART838 synergized strongly with BCL2 antagonists.Neither the detailed mechanism(s) of action nor the key molecular target(s) of artemisinins are known definitively, in cancers or microbes. Artemisinin activity seems to be mediated predominantly by generation of reduced oxygen species (ROS) by the endoperoxide pharmacophore(s) common to all artemisinins, leading to cell cycle arrest and apoptosis in cancer cell lines. Effects of artemisinins on levels of several pro-apopototic and anti-apoptotic molecules in cancer cell lines have been reported, including reduction of the BCL2 family member MCL1. MCL1 is frequently elevated in primary acute leukemias at initial diagnosis or at relapse, and MCL1 has been demonstrated as a mediator of resistance to BCL2 inhibitors in many AML and ALL cell lines. Therefore, we hypothesized that AS and ART838 might synergize with BCL2 inhibitors against acute leukemias by reducing the intracellular level and/or activity of MCL1.Levels of MCL1 protein, but not mRNA, were reduced in acute leukemia cell lines treated with AS or ART838. Artemisinin-mediated MCL1 reduction was selective, in that protein levels of other BCL2 family members were not affected by AS or ART838 treatment. When leukemia cells were treated with deoxy-ART838, an analog lacking the endoperoxide pharmacophore and inactive against either leukemias or malaria, caused much less MCL1 reduction. Co-treatment with AS or ART838 plus N-acetyl-cysteine (NAC) similarly decreased the reduction in levels of MCL1 protein. Thus, ROS appears necessary for the artemisinin-mediated post-transcriptional reduction of MCL1 protein levels.To confirm the potential role of MCL1 reduction in the observed antileukemic efficacy of artemisinins and in the observed synergy of artemisinins with ABT199, we used 3 established selective inhibitors of MCL1 as pharmacologic tools to cause loss-of-MCL1-function. S63845 and A1210477 selectively inhibit binding of BIM to MCL1, whereas maritoclax reduces cellular MCL1 levels by increasing degradation of MCL1 protein. We found that each of these 3 structurally dissimilar compounds added to the antileukemic efficacy of ABT199, but not AS, in AML and ALL cell lines, confirming the potential role for MCL1 reduction in the observed antileukemic efficacy of artemisinins and in the observed synergy of artemisinins with ABT199. Based on our observations, we propose the combination of AS or ART838 plus a BCL2 inhibitor such as ABT199 as a low-toxicity drug pair for incorporation into acute leukemia therapeutic regimens. DisclosuresFox:Intrexon Corporation: Employment, Equity Ownership. Tyner:Seattle Genetics: Research Funding; Syros: Research Funding; Janssen Pharmaceutica: Research Funding; Agios Pharmaceuticals: Research Funding; Takeda Pharmaceutical Company: Research Funding; Genentech: Research Funding; Incyte Corporation: Research Funding; Gilead: Research Funding; AstraZeneca: Research Funding; Array Biopharma: Research Funding; Leap Oncology: Consultancy; Constellation Pharmaceuticals: Research Funding; Aptose Biosciences: Research Funding. Civin:3DBioWork Inc: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees; BD (Becton Dickinson): Honoraria; ConverGene LLC: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding; GPB Scientific LLC: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees.

DNA Demethylating Agents Generate a Brcaness Effect in Multiple Sporadic Tumor Types: Prediction for Sensitivity to PARP Inhibitors in AML

Poly (ADP-ribose) polymerase inhibitors (PARPis) have shown clinical efficacy in breast and ovarian cancers with inherited mutations in BRCA genes that abrogate homologous recombination (HR) DNA double-strand break (DSB) repair activity. However, to date, PARPishave not had success in sporadic breast and ovarian cancers, or other cancers, such as acute myeloid leukemia (AML), with intact BRCA genes. We have recently reported that triple-negative breast cancer (TNBC) and AML are sensitive to low doses of the potent PARPiTalazoparib in combination with DNA methyl transferase inhibitors (DNMTi), decitabine (DAC) or azacitidine (AZA), in vitro and in vivo. In these cancer models, the DNMTi/PARPicombination increases amplitude and retention of PARP1 directly at laser-induced DNA damage sites, increasing DNA DSBs, promoting synergistic tumor cytotoxicity, blunting cell self-renewal, and leading to strong anti-tumor responses. These results have led to a Phase I/II Clinical trial of DNMTi/PARPi combination therapy in patients with relapsed/refractory AML that is underway.In identifying the molecular pathways that may underlie efficacy of this inhibitor combination in sporadic TNBC, ovarian cancer and AML, we find that low doses of DNMTisreprogram the cancer genome, in part by changing gene expression patterns of DSB repair genes, potentially creating a HR defect (HRD) or “BRCAness” effect. Expression array (Agilent Human 44K v2) analysis was performed on RNA extracted from multiple BRCA-intact sporadic cell lines from AML (including, Kasumi-1, KG-1 and MOLM-14), TNBC (including, MDA-MB-231, SUM159) and ovarian cancer (including, A2780) on days 1, 3, 5 and 7 and 10 post DNMTi(DAC 10nM or AZA 500nM) treatment. Hallmark, KEGG and reactome gene ontologies were utilized. Only gene sets with log fold change (FDR) >.250 were considered significant and genes demonstrating 2-fold or greater change with drug treatment were selected. While multiple DNA repair pathway genes were found both up- and down- regulated post DNMTi treatment, one or more HR genes, were consistently downregulated in all cell lines examined. QPCR of mRNA and/or immunoblotting of HR proteins, in particular RAD51, BRCA1, BRCA2, or HR-related genes, FEN1 or FANCD2, showed significantly decreased expression with DNMTi treatment (p<0.05) in TNBC, ovarian and AML cell lines, validating array data. Functional assays in TNBC cell lines, using a stably expressed HR repair reporter (DRGFP) also confirmed decreased HR activity with DNMTi treatment. Additionally, in response to DNA damage with ionizing radiation (IR, 4Gy), RAD51 foci were reduced in AZA-treated TNBC and ovarian cancer cell lines, compared to untreated controls. Significantly increased triradial chromosomes were also observed in TNBC in the presence of AZA when exposed to DNA cross-linking agents mitomycin C (MMC, 50ng/ml) or cisplatin (2μM) treatment, confirming functional down-regulation of the HR-related FANCD2 pathway.The above HRD-like or BRCAness phenotype with DNMTi treatment translated to sensitivity to PARPis in primary bone marrow mononuclear cells (BMMNC) from AML patients. Thus, in 6 of 8 (75%) AML samples, HR gene expression by QPCR of RNA and HR functional activity by immunostaining for RAD51 foci, following DAC treatment (10nM, 72 hrs), DNMTi treatment resulted in a significant decrease in HR or HR-related gene expression, including RAD51, BRCA1, BRCA2, and decreased levels of RAD51 foci (p<0.05). All 6 AML samples were also significantly more sensitive to DAC/talazoparib combination treatment (p<0.05) in colony forming assays, compared with single agent treatments. AML samples included those with MLL translocations that are reported to have high HR activity and resistance to PARPis. Studies are underway to elucidate the molecular characteristics of the minority of AML patient samples (N=2) that did not generate a BRCAness effect with DAC treatment. In conclusion, our data suggest that one of the mechanisms underlying DNMTi/PARPi therapy efficacy in AML is DNMTi-induced BRCAness. In vitro testing for BRCAness with DNMTitreatment could be a biomarker to predict sensitivity to DNMTi/PARPis and inform selection of AML patients for future clinical trials of this inhibitor combination. DisclosuresCivin:ConverGene LLC: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees, Research Funding; 3DBioWork Inc: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees; BD (Becton Dickinson): Honoraria; GPB Scientific LLC: Consultancy, Equity Ownership, Membership on an entity's Board of Directors or advisory committees. Kingsbury:3DBioWorks: Consultancy, Equity Ownership.

794. Healthcare-Associated Urinary Tract Infection (HA-UTI): A Risk Factor for Clostridioides difficile Infection (CDI)? Results of a Real-World Data Analysis

BackgroundIt is estimated that 223,900 cases of CDI occur annually in hospitalized patients resulting in 12,800 deaths and $1 billion in attributable costs. Antimicrobial use is a risk factor for CDI and the antimicrobials ordered to treat urinary tract infections have been identified as a factor in both recurrent CDI and community-acquired CDI. This real-world data analysis seeks to explore the relationship between HA-UTI and hospital-onset CDI (HO-CDI).MethodsAn electronic infection surveillance system was the source of de-identified real-world data from 290 hospitals. Algorithmically-derived measures of healthcare-associated infections (ADM-HAIs), and records of all-cause antimicrobial orders, for all inpatient admissions for the period 10/1/18–9/30/19 were analyzed. All patients who presented with a urine ADM-HAI – suggestive of HA-UTI – and no other healthcare-associated infection (Urine+ patients), were observed for subsequent HO-CDI. Urine+ patients were compared to patients with no HAI of any type, other than CDI (HAI-free patients), and relative risk (RR) was estimated. The analysis was repeated for the subgroup of patients who received an antimicrobial order for any reason during their stay.Results3,050,525 inpatient admissions were analyzed. 26,634 were identified as Urine+ patients. 188 of those patients subsequently presented with HO-CDI. 2,978,507 were identified as HAI-free patients. 6,238 of those patients presented with HO-CDI. The incidence of HO-CDI was significantly higher in Urine+ patients compared to HAI-free patients (RR=3.37, 95% CL[2.92, 3.89], p< 0.0001). When the analysis was repeated to examine only patients who received antimicrobial orders, Urine+ patients continued to be at higher risk of subsequent HO-CDI compared to HAI-free patients (RR=3.28, 95% CL[2.74,3.92], p< 0.0001).ConclusionThe presence of a urine ADM-HAI, suggestive of HA-UTI, was associated with an increased risk of subsequent HO-CDI. This held when only patients with antimicrobial orders were considered. These observations mirror findings from other published studies, however, other factors may have contributed to increased risk for both HA-UTI and HO-CDI.DisclosuresTimothy Kelly, MS, MBA, BD (Employee) ChinEn Ai, MPH, BD (Employee) John Murray, MPH, BD (Employee) Yan Xiong, n/a, BD (Becton Dickinson) (Employee) Hanna Jokinen-Gordon, PhD, BD (Employee)

Heterogeneity of Hematopoietic Stem and Progenitor Cell (HSPC) Composition in Αβ T-Cell/CD19 B-Cell Depleted Peripheral Blood Cell Stem Cell (PBSC) Transplant Grafts and Correlation with Immune and Hematopoietic Recovery

INTRODUCTION. While total CD34 counts in PBSC graft products have correlated with overall likelihood of hematopoietic recovery after allogeneic hematopoietic stem cell transplantation (HSCT), analyses of the HSPC composition and its relationship to relevant post-transplant clinical outcomes are lacking. In fact, the biological basis for different dynamics of hematopoietic/immune recovery, the risk of infection, and graft-versus-host disease (GvHD) is not fully understood. We have performed the first analysis of HSPC graft composition in 6 αβ T-cell/CD19 B-cell depleted haploidentical (αβhaplo) HSCT. Additionally, we correlated the HSPC graft composition with the distribution of the same HSPC subsets in serial post-HSCT bone marrow aspirates performed at Days 30, 60, and 90, with the peripheral blood counts [neutrophils, monocytes, platelet (PLT)] and with the immune recovery (CD3+, CD3+CD4+, CD3+CD8+, αβT, γδT, NK cells) at the same time points. The patients were divided in two groups: 3 patients had a robust and sustained hematopoietic recovery (Group 1) while 3 patients experienced mild cytopenia after Day 60 (Group 2). All patients were transplanted for acute leukemia and received a myeloablative TBI-based conditioning regimen. See Table 1 for details about patients and graft composition. METHODS. All patients were enrolled in the Stanford IRB approved BMT Protocols 179/351/361 and had peripheral blood (PB) and bone marrow (BM) evaluated at Day 30, 60 and 90 post HSCT for the primitive CD34+ Lin- HSPC subsets: HSC (CD38-CD45RA-CD90+), MPP (CD38+CD45RA+), CMP (CD38+CD45RA-CD123+), GMP (CD38+CD45RA+CD123+), MEP (CD38+CD45RA-CD123+) and CLP (CD38+CD127+). Aliquots of αβhaplo-HSCT grafts were cryopreserved for later analyses. Mononuclear cells were isolated from PBSC, PB and BM by Ficoll-Hypaque (Sigma-Aldrich) density gradient centrifugation. FACS analyses were performed on either fresh or frozen cells on Becton Dickinson (BD) Aria II flow cytometer. At least 5x105 events were acquired and analyzed using FlowJo software (BD). RESULTS. Despite consistent levels of αβT-cell depletion and CD34 enrichment, the frequency of the HSPC subsets varied between the grafts. Notably, while CMP and GMP were very consistent across the 6 grafts, the frequency of HSC, CLP, MEP and MPP showed a 2-fold range of variation (Fig1A). No significant correlation was observed between the HSC frequency in the graft and the hematopoietic/immune recovery. However, the frequency of HSC in the BM at Day 30 is statistically correlated (P=0.027) with the PLT at Day 90 (Fig1B). In these preliminary results, the different distribution of CMP, GMP, MEP and MPP did not impact on the hematopoietic/immune recovery. However, there was a significant correlation (P=0.02) between CLP and γδ T cells reconstitution at Day 90 in Group 1 patients (Fig1C). Additionally, the neutrophils, monocytes and phagocytic cells recovery at Day 90 is statistically correlated with the GMP frequency in the BM at Day 30 (P=0.017; P=0.018; P=0.0132, respectively) (Fig1D). Interestingly, the same strong correlation is observed between the CMP in the BM at Day 60 and the recovery of neutrophils and phagocytic cells (P=0.016, P=0.019) at Day 90 (Fig1E), but the CMP at day 30 are already predictive of a robust engraftment in the Group 1 patients (data not shown). CONCLUSIONS. In αβhaplo-HSCT, previously identified factors influencing hematopoietic recovery have been mainly limited to the enumeration of bulk CD34 counts and of mature effector cells, such as αβ/γδ T and NK cells. On the other hand, the presence of GvHD and thymic injury have been correlated to the kinetics of immune reconstitution. We hypothesized that the HSPC composition of the graft would impact lymphohematopoietic recovery in αβhaplo-HSCT recipients. Although preliminary, our data indicate that even with a consistent method of graft manipulation, the HSPC graft composition is heterogeneous. Variations in HSPC subsets frequency and number can contribute to significant differences in lymphohematopoietic recovery and, therefore, clinical outcome. The evaluation of a larger number of patients with longer follow up after HSCT are required. Comparative studies with unmanipulated T-cell replete and cord blood grafts are ongoing. Such analyses will be instrumental not only for prediction of clinical outcome, but also for optimization of novel graft engineering strategies. Disclosures No relevant conflicts of interest to declare.

Methicillin-resistant Staphylococcus aureus: epidemiology and antimicrobial susceptibility experiences from the University Hospital ‘Luigi Vanvitelli’ of Naples

ABSTRACT Methicillin-resistant Staphylococcus aureus (MRSA) is one of the important pathogens worldwide showing resistance to several widely used antibiotics. This has made the treatment of MRSA infections harder, especially due to their prevalence in the hospital setting. We evaluated the antibiotic susceptibility patterns of healthcare-associated MRSA infections with a focus on Vancomycin Intermediate S. Aureus (VISA) and macrolide-licosamide-streptogramin B (MLSB) phenotypes. A total of 417 Staphylococcus aureus (S. aureus) cases were isolated between January 2017 and December 2018, through several clinical specimens collected from the University Hospital ‘Luigi Vanvitelli’ of Naples. We identified bacterial strains using Matrix-Assisted Laser Desorption/Ionization-Time of Flight (MALDI-TOF) and antimicrobial susceptibility using Phoenix BD (Becton Dickinson, NJ, USA). Out of the total 417 S. aureus cases, 140 were MRSA (33.6%) and of these, 50% were soft tissue infections. All MRSA and Methicillin sensible S.aureus MSSA isolates were susceptible to linezolid and daptomycin. Two MRSA cases exhibited intermediate resistance to vancomycin and were of constitutive MLSB phenotype. Among the MRSA strains, 11.4% were constitutive and 43.6% were inducible MLSB phenotypes and 8.6% were macrolide-streptogramin B phenotype. This study characterized the epidemiological status, antibiotic resistance patterns, and current prevalent phenotypes of healthcare-associated MRSA. This knowledge can aid clinicians in improving the antimicrobial stewardship program by adapting appropriate guidelines for the proper use of MRSA antibacterial agents.

Comparison of the recommended colistin susceptibility testing methods with colistin gradient strips and semi-automated method for antimicrobial-resistant non-fermenting rods

An increased frequency of multidrug-resistant non-fermenting rods isolation has resulted in the excessive use of colistin – often the last chance antimicrobial. However, determination of colistin susceptibility is difficult, mainly because of its structure and limited diffusion properties. This study was performed to compare colistin susceptibility testing among Pseudomonas aeruginosa (n = 49) and Acinetobacter baumannii (n = 49) strains. Four methods were applied: colistin gradient strips (Liofilchem, Italy), semi-automated method Phoenix BD (Becton Dickinson, USA) and two broth microdilution methods: SensiTest Colistin (Liofilchem, Italy) and MICRONAUT MIC-Strip (MERLIN Diagnostika GmbH, Germany). Data were analyzed by comparison of MIC values and strains susceptibility interpretation criteria (resistant and sensitive, respectively).The same interpretation results were obtained for 46 (93.9%) P. aeruginosa and 37 (75.5%) A. baumannii isolates in all of the applied methods. Using broth microdilution methods, the same interpretation was obtained for 48 (98.0%) P. aeruginosa and 42 (85.7%) A. baumannii isolates.The results obtained by colistin gradient strips usually confirm the results of broth microdilution tests for P. aeruginosa isolates, the automated method is in turn less labor-intensive. However, MIC values, obtained with their use, are less precise because of the antibiotic dilutions limited to only several concentrations. The results underline the importance of choosing of the appropriate type of method, also among those recommended and commercially available.

Enumeration and characterization of airborne microbial communities in an outdoor environment of the city of Taj, India

In the present investigation, the microbiological characteristics of the air in the outdoor environment of the city of Taj, i.e., Agra (India) was assessed by determining the concentration and composition of bacteria and fungi using culture technique. The bacterial isolates having the highest prevalence (100%) throughout the year were selected and identified based on morphological, physiological, and biochemical characteristics. The mean concentration of bacteria in PM10 and PM2.5 was 405.1 CFUm-3 and 364.9 CFUm-3, respectively, while the mean concentration of fungi in PM10 and PM2.5 was 136.5 CFUm-3 and 20.3 CFUm-3, respectively. A new technique of BD (Becton Dickinson)BBL Crystal Mind Auto reader was used. The bacteria were identified as Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Micrococcus luteus, Chromobacterium violaceum, Bacillus cereus, Brevibacillus brevis, Enterobacter aerogenes, Pseudomonas aeruginosa, Acinetobacter beijerenckii. Fifteen fungal isolates were identified comprising of Fusarium, Aspergillus, Epicoccum, Penicillium, Cladosporium, Trichoderma. Alternaria, Mucor, Rhizopus, Geotrichum, Helminthosporium, Absidia, Cephalosporium, Botrytis, and Neurospora. Aspergillus and Penicillium have the maximum percentage occurrence of 98% and 95%, respectively. Botrytis recorded a minimum percentage of 22%, followed by Neurospora 20%. The concentration of bacteria and fungi depend upon the meteorological conditions.

A Randomized Trial Examining Plasma Exchange Using the Becton Dickinson (BD) Catheter In an Outpatient Apheresis Unit

A Randomized Trial Examining Plasma Exchange Using the Becton Dickinson (BD) Catheter In an Outpatient Apheresis Unit

Standardization of a compensation matrix for the use in an ISHAGE‐based single‐platform for CD34+ stem cell quantitation on the FACSVia™ flow cytometer

Becton Dickinson (BD) FACSVia™ is a recently developed flow cytometer with which worldwide clinical experience is limited. Recently, our center started using the single-platform BD Stem Cell Kit (BD SCE) for the quantitation of viable CD34+ (7-aminoactinomycin D-7-AAD-negative cells) on a BD FACSVia™. Currently, there are no formal recommendations for the fluorescence compensation of 7-AAD in this scenario. A 3-color fluorescence compensation matrix ("Optimized BD CS&T") was standardized based on the fluorescence of 7-AAD in samples of 27 individuals. The "Optimized BD CS&T" was compared with manual compensation (predicated method) and BD CS&T beads. The analysis of the viable CD34+ cells/mm3 showed a very strong correlation between the manual compensation versus "Optimized BD CS&T" (r=1.00) and manual versus BD CS&T (r=.99). The analysis of CD34+ viability (%) showed a very strong correlation for manual versus "Optimized BD CS&T" (r=.99). However, for manual versus BD CS&T, the correlation was inferior (r=.86). For viable CD34+ cells/mm3 , Bland-Altman plots showed a better concordance between manual and "Optimized BD CS&T" than between manual and BD CS&T. For CD34+ viability (%), the concordance was very good between manual and "Optimized BD CS&T", while there was a poor concordance between manual and BD CS&T. "Optimized BD CS&T" matrix proved to be more precise than the conventional BD CS&T beads. The "Optimized BD CS&T" matrix can be used along with BD SCE kit on BD FACSVia™ flow cytometers.

2161. Organism-Specific Turn Around Time Improvement in Urinary Specimens as a Result of Microbiological Laboratory Automation

BackgroundUniversity Medical Center in Lubbock, TX is one of few medical centers using Becton Dickinson (BD) Kiestra Total Laboratory Automation (TLA) system since May 2015. The impact on organism-specific turn around time (TAT) in urinary specimens after implementation of TLA was evaluated.MethodsAfter approval from the Quality Improvement Review Board, a retrospective analysis of microbiological data from urinary specimens in BD research database was performed. Before vs. after implementation (2013 vs. 2016) TAT was compared. Ten clinically relevant organisms were analyzed. Statistical analysis was performed with SAS software version 9.2. Data were analyzed using Chi-squared test. A P-value of < 0.05 was considered statistically significant.ResultsOverall, 2282 specimens from 2013 and 2306 specimens from 2016 were analyzed. Compared with before vs. after implementation of TLA, an overall improvement in TAT was observed (expressed as mean hours for each organism): Enterococcus faecalis (55.2 vs. 38.8), Enterococcus faecium (68.4 vs. 43.8), Escherichia coli (44.2 vs. 41.0), Klebsiella pneumoniae (45.0 vs. 44.0), Proteus mirabilis (44.8 vs. 38.6), Pseudomonas aeruginosa (58.9 vs. 37.7), Staphylococcus aureus (49.2 vs. 36.0), Streptococcus agalactiae (49.2 vs. 31.4), Streptococcus pneumoniae (51.7 vs. 61.8), Streptococcus pyogenes (62.6 vs. 26.6). It was also observed that improvement in TAT was more pronounced for Gram-positive organisms than Gram-negative organisms. P-value was < 0.01 for all organisms except Streptococcus pneumoniae (0.7985) and Streptococcus pyogenes (0.2562). The number of specimens with these two organisms was too small to be considered significant.ConclusionAutomation of microbiology laboratory leads to significant TAT improvement in urinary specimens, making early data availability to clinicians. This improves efficiency as well as supporting earlier antibiotic switch, antimicrobial stewardship and optimal patient care in treating urinary tract infections.Disclosures All authors: No reported disclosures.

1731. Immune Dysregulation in Mucormycosis

BackgroundMucormycosis is a fatal fungal infection with unique predisposition to infect diabetics. Dysregulated adaptive immunity contributes to the pathogenesis in all fungal diseases, but activated Th17 cells have laid a new dimension to chronic inflammatory response which was previously attributed to uncontrolled Th1 response. We attempted to study the Th17 and T regulatory (Treg) immune response in diabetic patients with mucormycosis and compared the data with a healthy control and a T2DM case without fungal infection. In addition we could follow-up one patient post 6-month treatment and performed immunological studies.Methods2 mL of blood samples were collected in EDTA vial from two patients who were suffering from diabetes with mucormycosis for immunological investigations. Samples were also taken from age-matched T2DM patient without fungal infection and a healthy volunteer as controls for T-cell parameters. Repeat blood sample was taken to study immune parameters in one patient who was followed up after 6 months. The expression of various T-cell markers was analyzed by immunostaining with the antibodies against CD3, CD4, CD25, CD161, IL-23R [Becton Dickinson (BD) PharMingen]. Fluorescence profiles were analyzed using Flow Jo software (BD Biosciences). The results are expressed as a percentage of positive cells.ResultsThe percentages of CD4+ cells were low in both patients when compared with and healthy control but it is much higher in diabetes case when compared with others. CD161+ cell population was higher in both patients when compared with healthy control and diabetic patient without fungal infection. The percentage of IL23R+ cells was significantly high in patient before treatment when compared with, healthy control and diabetics. and decline after treatment. The percentage positivity of CD25+ cells was highest in healthy control when compared with others. The profile of CD25+ cells was comparatively similar in patient before treatment and diabetics but we found a higher percentage, in patients after treatment.ConclusionThe findings in this study imminently indicate the mechanism of immune dysregulation involving Th17 and Treg pathways in mucormycosis and provide evidence that restoration of Th17/Treg may be considered as a therapeutic option for long-term benefit in diabetics.Disclosures All authors: No reported disclosures.

Release of silicone oil and the off-label use of syringes in ophthalmology

Background/aimsTo assess silicone oil (SO) release by different brands of syringes used for intravitreal injection under different handling conditions.MethodsEight syringes were analysed: from the USA, Terumo 0.5 mL, Becton-Dickinson (BD)...