Early Predictors of Ventilator Associated Pneumonia in Preterm Neonates Admitted in a Special Newborn Care Unit.

OBJECTIVES: To determine the utility of microscopic examination and culture of endotracheal aspirate (ETA) in the early diagnosis of ventilator-associated pneumonia (VAP) in preterm neonates. METHODS: We enrolled 80 consecutive neonates (both inborn and out-born) with gestational age of < 37 weeks admitted in Special Newborn Care Unit (SNCU) and requiring mechanical ventilation (MV) for ≥ 48 hours. The diagnosis of VAP was made using the criteria laid down by the Centers for Disease Control (CDC). RESULTS: 47 preterm neonates (58.5%) developed VAP; the overall incidence was 74.7/1000 ventilator-days. The mean (SD) time (hours) to ETA culture was less as compared to diagnosis based on CDC criteria [108.9 (8.00 hrs) vs 132.4 (53.24); P = 0.004] with sensitivity and specificity of 80.8% and 72.7%, respectively. Outborn delivery was the single most important risk factor for VAP. Multidrug resistant (MDR) Klebsiella pneumoniae (63.9%) was the most prevalent organism. CONCLUSIONS: We noticed a very high incidence of VAP among preterm neonates in SNCU. ETA culture can aid in early diagnosis.

Synthesis and studies of new purines/pyrimidine derivatives as multi-targeted agents involving various receptor sites in the immune system.

Pro-inflammation, which is developed due to the increased production of cytokines, mainly interleukin-6 (IL-6), during the working of immune system pathways, becomes a major concern these days for many researchers. So, it is desired to design, screen, and synthesize new molecules with multi-parametric features showing their efficacy for Toll-like receptors (TLRs) and inhibiting the disease-causing receptor sites like viral infections, cancers, etc. along with controlling inflammation, fever, and other side effects during such pathways. Further, looking at the literature, curcumin a multi-targeted agent is showing its efficiency toward various receptor sites involved in many diseases as mentioned above. This fascinated us to build up new molecules which behave like curcumin with minimum side effects. In silico studies, involving ADMET studies, toxicological data, and docking analyses, of newly synthesized compounds (3-5) along with tautomers of curcumin i.e., (1-2), and some reported compounds like 9 and 10 have been studied in detail. Great emphasis has been made on analyzing binding energies, protein-ligand structural interactions, stabilization of newly synthesized molecules against various selected receptor sites using such computational tools. Compound 3 is the most efficient multifunctional agent, which has shown its potential toward most of the receptor sites in docking analysis. It has also responded well in Molecular dynamics (MD) simulation toward 5ZLN, 4RJ3, 4YO9, 4YOJ, and 1I1R sites. Finally, studies were extended to understand in vitro anti-inflammatory activity for particularly compound 3 in comparison to diclofenac and curcumin, which signifies the efficiency of compound 3.

Investigating multifaceted action of ibuprofen derivatives towards cox isozymes and interleukin-6 (IL-6) site correlating with various target sites.

The multifaceted action of new ibuprofen analogs has been investigated against inflammation, neurological and pro-inflammation factors. On the basis of ADMET (absorption, distribution, metabolism, excretion, and toxicity) analysis, molecular docking as well as molecular dynamics simulation, compound 3 was thought to have good anti-inflammatory activity. As the presence of structural interactions such as conventional hydrogen bonds and electrostatic interactions through the nitrogen atoms of the linker in compound 3 gave strong evidence of its potency. The major finding of the current work is that the presence of appropriate number of hetero atoms (NH, OH) in a compound makes it more efficient than the number of labile groups (i.e., hydroxyl groups). Additionally, the position of hetero atoms in a compound and orientation also play a vital role in its efficacy. It was also screened for in vitro anti-inflammatory activity by membrane stability method, where it has shown 90.8% protection of RBC hemolysis. Thus, compound 3 with effective structural features may have good anti-inflammatory activity.Communicated by Ramaswamy H. Sarma.

Does the Capsule Method Increase the Accuracy of the Detection of Ger by Scintigraphy Technique?

Background: The scintigraphy technique is the most sensitive test for the detection of gastroesophageal reflux disease (GERD). Scintigraphy techniques employ two methods: the liquid method and the capsule method. Aim: In this prospective study, we are trying to find out the efficiency of the capsule method for gastroesophageal reflux scintigraphy over the liquid method and to determine the ease of execution of the technique and the interpretation of the results. Materials and Methods: A total of 65 symptomatic patients (age range: 7-71 years; mean age: 35.2 years) were included in the study. They were divided into two groups: group A, which included 18 patients who underwent the liquid method and Group B, which included 47 patients who underwent the capsule method. The average administered dose of 99mTc-labeled sulfur colloid was 11.1-18.5 MBq. Results: The results showed that 45 (69.12%) of the 65 patients tested positive for GERD. Furthermore, 15 were positive in the liquid method and 30 in the capsule method. Grade III reflux was seen in 66.67% of patients, and 33.33% of patients with Grade II and I reflux were diagnosed using both methods. Conclusion: Thus, in conclusion, we can say that both liquid and capsule methods are equally sensitive for the detection of low as well as high refluxate volumes.

Multiepitope glycan based laser assisted fluorescent nanocomposite with dual functionality for sensing and ablation of Pseudomonas aeruginosa.

Pseudomonas aeruginosa (P. aeruginosa) infection is becoming a severe health hazard and needs early diagnosis with high specificity. However, the non-specific binding of a biosensor is a challenge to the current bacterial detection system. For the first time, we chemically synthesized a galactose tripod (GT) as a P. aeruginosa-specific ligand. We conjugated GT to a photothermally active fluorescent nanocomposite (Au@SiO2-TCPP). P. aeruginosa can be detected using Au@SiO2-TCPP-GT, and additionally ablated as well using synergistic photothermal and photodynamic therapy. Molecular dynamics and simulation studies suggested better binding of GT (binding energy = -6.6 kcal mol-1) with P. aeruginosa lectin than that of galactose monopod (GM) (binding energy = -5.9 kcal mol-1). Furthermore, a binding study was extended to target P. aeruginosa, which has a galactose-binding carbohydrate recognition domain receptor. The colorimetric assay confirmed a limit of detection (LOD) of 104 CFU mL-1. We also looked into the photosensitizing property of Au@SiO2-TCPP-GT, which is stimulated by laser light (630 nm) and causes photoablation of bacteria by the formation of singlet oxygen in the surrounding media. The cytocompatibility of Au@SiO2-TCPP-GT was confirmed using cytotoxicity assays on mammalian cell lines. Moreover, Au@SiO2-TCPP-GT also showed non-hemolytic activity. Considering the toxicity analysis and efficacy of the synthesized glycan nanocomposites, these can be utilized for the treatment of P. aeruginosa-infected wounds. Furthermore, the current glycan nanocomposites can be used for bacterial detection and ablation of P. aeruginosa in contaminated food and water samples as well.

Drought stress detection technique for wheat crop using machine learning.

The workflow of this research is based on numerous hypotheses involving the usage of pre-processing methods, wheat canopy segmentation methods, and whether the existing models from the past research can be adapted to classify wheat crop water stress. Hence, to construct an automation model for water stress detection, it was found that pre-processing operations known as total variation with L1 data fidelity term (TV-L1) denoising with a Primal-Dual algorithm and min-max contrast stretching are most useful. For wheat canopy segmentation curve fit based K-means algorithm (Cfit-kmeans) was also validated for the most accurate segmentation using intersection over union metric. For automated water stress detection, rapid prototyping of machine learning models revealed that there is a need only to explore nine models. After extensive grid search-based hyper-parameter tuning of machine learning algorithms and 10 K fold cross validation it was found that out of nine different machine algorithms tested, the random forest algorithm has the highest global diagnostic accuracy of 91.164% and is the most suitable for constructing water stress detection models.

Nanoparticle-Shielded dsRNA Delivery for Enhancing RNAi Efficiency in Cotton Spotted Bollworm Earias vittella (Lepidoptera: Nolidae).

The spotted bollworm Earias vittella (Lepidoptera: Nolidae) is a polyphagous pest with enormous economic significance, primarily affecting cotton and okra. However, the lack of gene sequence information on this pest has a significant constraint on molecular investigations and the formulation of superior pest management strategies. An RNA-seq-based transcriptome study was conducted to alleviate such limitations, and de novo assembly was performed to obtain transcript sequences of this pest. Reference gene identification across E. vittella developmental stages and RNAi treatments were conducted using its sequence information, which resulted in identifying transcription elongation factor (TEF), V-type proton ATPase (V-ATPase), and Glyceraldehyde -3-phosphate dehydrogenase (GAPDH) as the most suitable reference genes for normalization in RT-qPCR-based gene expression studies. The present study also identified important developmental, RNAi pathway, and RNAi target genes and performed life-stage developmental expression analysis using RT-qPCR to select the optimal targets for RNAi. We found that naked dsRNA degradation in the E. vittella hemolymph is the primary reason for poor RNAi. A total of six genes including Juvenile hormone methyl transferase (JHAMT), Chitin synthase (CHS), Aminopeptidase (AMN), Cadherin (CAD), Alpha-amylase (AMY), and V-type proton ATPase (V-ATPase) were selected and knocked down significantly with three different nanoparticles encapsulated dsRNA conjugates, i.e., Chitosan-dsRNA, carbon quantum dots-dsRNA (CQD-dsRNA), and Lipofectamine-dsRNA conjugate. These results demonstrate that feeding nanoparticle-shielded dsRNA silences target genes and suggests that nanoparticle-based RNAi can efficiently manage this pest.

Salmonella typhimurium detection and ablation using OmpD specific aptamer with non-magnetic and magnetic graphene oxide.

Foodborne diseases have increased in the last few years due to the increased consumption of packaged and contaminated food. Major foodborne bacteria cause diseases such as diarrhea, vomiting, and sometimes death. So, there is a need for early detection of foodborne bacteria as pre-existing detection techniques are time-taking and tedious. Aptamer has gained interest due to its high stability, specificity, and sensitivity. Here, aptamer has been developed against Salmonella Typhimurium through the Cell-Selex method, and to further find the reason for specificity and sensitivity, OmpD protein was isolated, and binding studies were done. Single molecular FRET experiment using aptamer and graphene oxide studies has also been done to understand the mechanism of FRET and subsequently used for target bacterial detection. Using this assay, Salmonella Typhimurium can be detected up to 10 CFU/mL. Further, Magnetic Graphene oxide was used to develop an assay to separate and ablate bacteria using 808 nm NIR where temperature increase was more than 60 °C within 30 s and has been shown by plating as well as a confocal live dead assay. Thus, using various techniques, bacteria can be detected and ablated using specific aptamer and Graphene oxide.

Derivatized chitosan-oil-in-water nanocapsules for trans-cinnamaldehyde delivery.

trans-Cinnamaldehyde, known for its bacterial anti-quorum sensing activity when applied at sublethal concentrations, has gained traction given its potential use against multidrug resistant bacteria. In this work, trans-cinnamaldehyde-loaded oil-in-water nanocapsules coated with chitosan, N,N,N-trimethyl chitosan chloride, N-(2-(N,N,N-trimethylammoniumyl)acetyl) chitosan chloride or N-(6-(N,N,N-trimethylammoniumyl)hexanoyl)chitosan chloride were obtained. All the formulated nanocapsules showed a Z-average hydrodynamic diameter ~ 160 nm and ζ-potential higher than +40 mV. N,N,N-trimethyl chitosan-coated oil-in-water nanocapsules showed the greatest trans-cinnamaldehyde association efficiency (99.3 ± 7.6) % and total payload release (88.6 ± 22.5) %, while N-(6-(N,N,N-trimethylammoniumyl)hexanoyl)chitosan chloride chitosan-coated oil-in-water nanocapsules were the only formulations stable in phosphate buffer saline PBS (pH 7.4) upon incubation at 37 °C for 24 h. Future work should address the stability of the developed nanocapsules in culture media and their biological performance.

Switching of newly synthesized linker-based derivatives of non-steroidal anti-inflammatory drugs toward anti-inflammatory and anticancer activity.

A new series of linker-based derivatives of non-steroidal anti-inflammatory drugs were designed and synthesized. All the compounds were well characterized with the help of various spectroscopic techniques such as FT-IR, 1H NMR, 13C NMR, and HRMS. The main emphasis of this paper is to understand the switching of the most promising compounds 8 and 10 towards anti-inflammatory and anticancer activity in terms of in-silico and in-vitro studies in detail. During the molecular docking study, compounds 8 and 10 demonstrated the importance of hetero atoms as well as the perfect alignment of a compound in the binding pocket of a target site, which may affect their bioactivity. Here, the presence of 1,3­dicarbonyl interactions with ASN 351 in compound 8 (not found in compound 10) may be responsible for its better inhibitory activity against the COX-2 target site. On the other hand, a slight increase in the potency of compound 10 towards anticancer activity may be due to the instantaneous participation of the OH group and carbonyl group to give conventional hydrogen bonds towards THR 149 amino acid residue, which was missing in compound 8. Molecular dynamics simulation was also performed for compounds 10 and 8 toward COX-2 and HER-2 protein sites. Further, compounds 8 and 10 were subjected to in-vitro COX-2 inhibition and cytotoxicity assay and the results obtained were in accordance with the in-silico study. Thus, compound 8 become more potent towards COX-2 inhibition with IC50 value of 48.51 µg/ml and compound 10 showed good bioactivity toward cytotoxic activity with IC50 value of 93.03 µg/ml.

Doublesex homolog is sex-specifically spliced and governs the sexual differentiation process in the whitefly Bemisia tabaci AsiaII-1.

The silverleaf whitefly Bemisia tabaci is one of the most destructive of crop pests globally. In Northern India cotton is predominately infested by the Asia II-1 species of B. tabaci. Though B. tabaci exhibits unique haplodiploidy in its reproductive behavior, to date very little is known regarding its sex determination mechanism. Here, an in-depth characterization of the AsiaII-1 doublesex (Btdsx) gene, which has been implicated in sex determination in B. tabaci, indicates the inclusion of six exons and five introns. The pre-mRNA is shown to sex-specifically splice, producing four male isoforms and one female isoform. These BtDsx proteins share common DNA binding (OD1) domains whereas they differ at their C-termini. RT-qPCR analysis revealed a significantly higher expression of Btdsx in female adults compared to that in male adults and earlier developmental stages. Functional characterization of Btdsx through RNA interference (RNAi) resulted in a significant reduction in its expression in both sexes. Btdsx knockdown concomitantly resulted in up-regulation of the expression of vitellogenin (vg) and vitellogenin receptor (vgr) genes in males and their down-regulation in females. Btdsx knockdown followed by mating resulted in reduced fecundity and percent egg hatching; however, no impact was observed on the female: male ratios in the progeny obtained from knockdown parents.

Inline imaging reveals evolution of the size distribution and the concentration of microbubbles in dissolved air flotation.

Dissolved air flotation (DAF) is an efficient process to remove impurities from fresh or salt water. As the removal is based on the agglomeration of impurities on the generated microbubbles, the size distribution and concentration of air bubbles are key parameters in dissolved air flotation. However, the development of microbubbles in the whole flotation process remains unexplored. In this study, we show that state-of-the-art inline microscopy enables the image acquisition of bubbles in DAF. Based on image analysis, thousands of microbubbles (10-200 µm) were analyzed within 6-12 min experiments. Consequently, bubble size distributions and bubble concentrations can be determined with moderate effort. Bubble size distributions were measured in a lab-scale DAF comprising a saturation unit, a decompression valve in/after which the bubbles are formed, and the actual flotation tank. The state of the microbubbles is not only determined at different positions within the tank but also in the supply pipe from the decompression valve to the tank. All bubble size distributions were unimodal and can be described well with Burr XII distributions. For fresh water, bubble size increased while bubble concentration decreased along the supply pipe between the decompression valve and the inlet of the flotation tank, indicating bubble coalescence. Compared to freshwater, saltwater inhibited this bubble coalescence in the pipe. Within the flotation tank, the bubble size did not change drastically for neither salt- nor freshwater. However, the bubble concentration decreased for both waters, which could be explained by dilution effects. Our results demonstrate that the developed inline method is a promising tool to study the evolution of microbubbles in flotation systems. Further, it might also be applied to investigate microbubbles in other processes such as fermentation, decomposition of organic compounds, and fouling mitigation in membranes.

Job satisfaction among medical officers working in Delhi.

Background: Satisfied human resource is an essential asset for high productivity, efficiency, and quality of care in a healthcare organization. Job satisfaction among healthcare professionals assumes further significance for the purpose of maximization of human resource potential in the context of the shortage of manpower in the health sector in India. Objectives: The objective of the study was to assess the job satisfaction among medical officers (MOs) working in Delhi and to study the factors influencing the satisfaction level amongst them. Material and Methods: It was a cross-sectional study conducted amongst selected MOs (by convenience) in all the districts of Delhi. A structured, self-administered questionnaire was used to collect data from the participants. Results: A total of 115 MOs completed the questionnaire. There were 61 (53%) female doctors and 54 (47%) male doctors. The mean age of the study participants was 38.28 ± 7.37 years. The experience of the study group ranged between 1 and 28 years (9.59 ± 6.47 years). Of the total, 59.1% of participants were found to be satisfied with their job. The most common reason given for satisfaction with job was the timing of their job that suits them. The reason given for not being satisfied with job was salary not at par with work pressure. A significantly higher proportion of male participants considered "lack of accommodation" (P < 0.03) and "frequent detailment" (P < 0.031) are big problems faced in their jobs as compared with the female participants. Only 37 (32.2%) participants felt that the MO is able to give enough time to patients for clinical assessment in Out Patient Department. Conclusions: Although majority (59%) of the study group was satisfied with their job, there were numerous problems faced by the doctors at their workplace. Low pay was one of the important reasons cited for being dissatisfied. There is an urgent need to address all the factors affecting job satisfaction through proper planning and implementing relevant human resource policies for improvement in the work environment at public health institutions.

Inferring Agronomical Insights for Wheat Canopy Using Image-Based Curve Fit K-Means Segmentation Algorithm and Statistical Analysis.

Phenomics and chlorophyll fluorescence can help us to understand the various stresses a plant may undergo. In this research work, we observe the image-based morphological changes in the wheat canopy. These changes are monitored by capturing the maximum area of wheat canopy image that has maximum photosynthetic activity (chlorophyll fluorescence signals). The proposed algorithm presented here has three stages: (i) first, derivation of dynamic threshold value by curve fitting of data to eliminate the pixels of low-intensity value, (ii) second, extraction and segmentation of thresholded region by application of histogram-based K-means algorithm iteratively (this scheme of the algorithm is referred to as the curve fit K-means (CfitK-means) algorithm); and (iii) third, computation of 23 grey level cooccurrence matrix (GLCM) texture features (traits) from the wheat images has been done. These features help to do statistical analysis and infer agronomical insights. The analysis consists of correlation, factor, and agglomerative clustering to identify water stress indicators. A public repository of wheat canopy images was used that had normal and water stress response chlorophyll fluorescence images. The analysis of the feature dataset shows that all 23 features are proved fruitful in studying the changes in the shape and structure of wheat canopy due to water stress. The best segmentation algorithm was confirmed by doing exhaustive comparisons of seven segmentation algorithms. The comparisons showed that the best algorithm is CfitK-means as it has a maximum IoU score value of 95.75.

Identification and validation of stage-specific reference genes for gene expression analysis in Callosobruchus maculatus (Coleoptera: Bruchidae).

In light of a number of recent studies highlighting the increasing research interest in bruchids, it is crucial to validate suitable reference genes that could be used in quantitative gene expression studies. Callosobruchus maculatus is a serious pest of stored grains and field legumes in which reference genes have not been assessed and validated to date. The present study aimed to identify and validate reference genes in different developmental stages of C. maculatus shortlisted from commonly used reference genes such as VATPase, TRIP12, TBP, TF11D, ACTIN, GST, ANNEXIN, PTCD3, RPL32, and ß -Tub in various insects. Dedicated algorithms like GeNorm, NormFinder, and BestKeeper were used to analyze the stability of these candidate genes, which revealed GST for third instar, ANNEXIN and PTCD3 for the fourth instar, TF11D and VATPase for male pupa, RPL32 and ß-tub for female pupa, ß-tub and TBP for adult male and VATPase and GST for adult females as suitable reference genes for expression studies in C. maculatus. The final comprehensive ranking using RefFinder identified GST and TBP as the best reference genes for all the developmental stages of C. maculatus. To the best of our knowledge, this is the first report which evaluates and validates stable reference genes in C. maculatus. The information of stage-specific gene expression, generated in this study will be useful for future molecular, physiological, and biochemical studies on C. maculatus and other closely related bruchids.

Correlation of Vitamin D Deficiency with Predictors of Mortality in Critically Ill Children at a Tertiary Care Centre in North India-A Prospective, Observational Study.

This study was conducted to assess the relationship of vitamin D deficiency (VDD) with various demographic characteristics, laboratory parameters, and predictors of mortality. This prospective observational study was performed at pediatric intensive care unit (PICU) of a tertiary care hospital situated in north India. A total of 125 children admitted in PICU with age from 2 months to 14 years were analyzed. The subjects were classified as Vitamin D deficient (≤20 ng/mL) and nondeficient (>20 ng/mL). The relationship between VDD and predictors of mortality were analyzed using correlation and multivariate analysis. Respiratory system (40%) was most commonly involved. VDD was seen in 72% of the patients. There was statistically significant correlation of VDD with age ( p = 0.019), season ( p = 0.018), height ( p = 0.005), and weight ( p = 0.003). On multivariate analysis factors associated with VDD were age (odds ratio [OR] = 1.01, 95% confidence interval [CI] 1.00-1.03, p = 0.006), season (OR = 3.98, 95% CI 1.09-14.50, p = 0.036). VDD was also correlated to bacteriuria ( p = 0.033), cardiovascular sequential sepsis-related organ failure assessment score (CV-SOFA score) ( p = 0.001), and mechanical ventilation ( p = 0.043). On multivariate analysis, factors associated with VDD were bacteriuria (OR = 4.88, 95% CI 1.04-22.89, p = 0.04), mechanical ventilation requirement (OR = 2.95, 95% CI 1.12-7.85, p = 0.029), and CV-SOFA score (OR = 2.33, 95% CI 1.14-4.76, p = 0.021). Median (interquartile range) duration of PICU stay in VDD patients was (3-7) days while in nondeficient patients it was (2-6) days ( p = 0.107). VDD was a significant risk factor for the need of mechanical ventilation, bacteriuria, and mortality among patients in our cohort.

Evaluation of antenatal services at Family welfare Centre under RMNCH+A Programme in Delhi.

BACKGROUND: According to WHO, 536,000 women die every year in the world from causes relating to pregnancy, childbirth, or postpartum. Ninety nine percent of these deaths occur in the developing countries. Primary health centres (in densely populated areas and in urban slums are also referred to as family welfare centres, since they provide the whole continuum of care, from birth through adolescent and delivery and thereafter. AIM: To evaluate the antenatal services under Reproductive, Maternal, Newborn, Child plus Adolescent (RMNCHA) programme at a family welfare centre located in Central Delhi, India. We have proposed an action plan that will help in evaluating and improving these services. METHODOLOGY: The study included health providers of antenatal services and the beneficiaries. With a response rate of 90% from a sample of 218, 203 consenting antenatal women (beneficiaries) visiting the centre for ANC check-up under RMNCH + A programme and PMSMA were included. For quantitative component, a pre-designed, pre-tested semi-structured screening questionnaire were administered to the beneficiaries and healthcare providers to evaluate the antenatal services under RMNCH + A programme at the family welfare centre. RESULTS: Qualitative and quantitative analysis was done separately. Only 43% of ANC women said health workers visited them at home during pregnancy and very few could tell about importance of lab investigations. Almost all (97%) knew about key messages given by health workers. Nearly all participants considered that the screening process was smooth. The waiting time and time to undergo varied from 4 to 6 min.

Quality Improvement Initiative to Improve Hand Hygiene Compliance in Indian Special Newborn Care Unit.

Pilot data on Hand hygiene (HH) compliance using a standard World Health Organisation checklist for 1-week suggested only 20% compliance. So, we planned a Quality Improvement study to improve HH compliance among health care providers in our Special Newborn Care Unit from 20% to 60% over 12 months. METHODS: We did this study in 3 phases: Baseline phase (2 months), Intervention phase (8 months), and Postintervention phase (2 months). A multidisciplinary Quality Improvement team composed of doctors, nursing staff, and ward attendants was constituted. The team analyzed potential barriers to HH by Fishbone analysis. Three trained observers randomly selected two target Special Newborn Care Unit patients daily and collected data on HH compliance unobtrusively during the three 8-h shifts over 24 h. In addition, we tested a range of interventions using multiple Plan Do Study Act cycles: Staff education; Displaying posters; Round the clock availability of soap and hand rub; Staff felicitation; Group performance feedback. We also collected data on healthcare-associated infections in all three phases. RESULTS: The total observations for HH during the baseline, intervention, and postintervention phase were 1488, 5808, and 1464, respectively. The HH compliance improved from 27.2% to 57.1% in the postintervention phase. There was no difference in the healthcare-associated infections among the three phases. CONCLUSIONS: The HH compliance rates improved significantly but not to the desired extent. So, we planned to increase our workforce, and improve our training program and infrastructure.

Complex Tumor Spheroids, a Tissue-Mimicking Tumor Model, for Drug Discovery and Precision Medicine.

Malignant tumors are complex tissues composed of malignant cells, vascular cells, structural mesenchymal cells including pericytes and carcinoma-associated fibroblasts, infiltrating immune cells, and others, collectively called the tumor stroma. The number of stromal cells in a tumor is often much greater than the number of malignant cells. The physical associations among all these cell types are critical to tumor growth, survival, and response to therapy. Most cell-based screens for cancer drug discovery and precision medicine validation use malignant cells in isolation as monolayers, embedded in a matrix, or as spheroids in suspension. Medium- and high-throughput screening with multiple cell lines requires a scalable, reproducible, robust cell-based assay. Complex spheroids include malignant cells and two normal cell types, human umbilical vein endothelial cells and highly plastic mesenchymal stem cells, which rapidly adapt to the malignant cell microenvironment. The patient-derived pancreatic adenocarcinoma cell line, K24384-001-R, was used to explore complex spheroid structure and response to anticancer agents in a 96-well format. We describe the development of the complex spheroid assay as well as the growth and structure of complex spheroids over time. Subsequently, we demonstrate successful assay miniaturization to a 384-well format and robust performance in a high-throughput screen. Implementation of the complex spheroid assay was further demonstrated with 10 well-established pancreatic cell lines. By incorporating both human stromal and tumor components, complex spheroids might provide an improved model for tumor response in vivo.