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The researcher aims to evaluate the effect topical application of breastmilk on separation time of umbilical cord among newborns. It was observed that infection is a major cause of newborn mortality, and can be caused by bacteria entering the body through a newly-cut umbilical cord. Each year, approximately four million neonatal deaths occur globally and infections account for 36% of these deaths. According to WHO, antibacterial factors are present in human milk. WHO has included breast milk application to the cord as harmless practices.1 So, it was concluded that there is association between topical application of breast milk on umbilical cord and separation time of umbilical cord.

Human haematopoietic progenitor/stem cells (HPCs) differentiate into functional T cells in the thymus through a series of checkpoints. A convenient in vitro system will greatly facilitate the understanding of T-cell development and future engineering of therapeutic T cells. In this report, we established a lentiviral vector-engineered stromal cell line (LSC) expressing the key lymphopoiesis regulator Notch ligand, Delta-like 1 (DL1), as feeder cells (LSC-mDL1) supplemented with Flt3 ligand (fms-like tyrosine kinase 3, Flt3L or FL) and interleukin-7 for the development of T cells from CD34(+) HPCs. We demonstrated T-cell development from human HPCs with various origins including fetal thymus (FT), fetal liver (FL), cord blood (CB) and adult bone marrow (BM). The CD34(+) HPCs from FT, FL and adult BM expanded more than 100-fold before reaching the beta-selection and CD4/CD8 double-positive T-cell stage. The CB HPCs, on the other hand, expanded more than 1000-fold before beta-selection. Furthermore, the time required to reach beta-selection differed for the various HPCs, 7 days for FT, 14 days for FL and CB, and 35 days for adult BM. Nevertheless, all of the T cells developed in vitro were stalled at the double-positive or immature single-positive stage with the exception that some CB-derived T cells arrived at a positive selection stage. Consequently, the LSC-mDL1 culture system illustrated diverse T-cell development potentials of pre- and post-natal and adult human BM HPCs. However, further modification of this in vitro T-cell development system is necessary to attain fully functional T cells.

Background: Recognizing factors that cause prehospital stroke delays may improve time of presentation to the Emergency Department (ED) and allow earlier treatment of acute stroke patients. Study Objectives: To determine the impact of stroke recognition by emergency medical dispatchers (EMD) and paramedics (PM) on ED arrival time in a large urban Emergency Medical Services system. Methods: Retrospective study of patients aged 18 years or more identified as having acute stroke by EMD, PM, or stroke neurologists from January 1, 2005 to December 31, 2005. Data were acquired from computer-assisted dispatch records, paramedic assessments, ICD-9 (International Classification of Diseases, 9 th Revision) databases, and a hospital stroke registry. Paramedic time to scene, scene time, and total run time were computed for patients with final hospital diagnosis of stroke and grouped into missed strokes and identified strokes by EMD and PM. Time intervals were compared between missed and identified strokes as well as between incidents where EMD and PM agreed or disagreed. Results: A total of 1067 patients were eligible for the study; 22 were excluded for missing data. For true strokes, EMD and PM were in agreement 27.3% of the time. The median RT was 2.5 min shorter when there was agreement between the providers than when there was disagreement (36.5 min; interquartile range [IQR] 30–43 vs. 39 min.; IQR 33–45, respectively). Conclusions: Prehospital scene time and run times for acute strokes are less when there is diagnostic concordance between dispatchers and paramedics. Time intervals did not differ between missed and recognized strokes.

Objectives: The aim of the investigation was to use in vitro transposon mutagenesis to generate metronidazole resistance in the obligately anaerobic pathogenic bacterium Bacteroides thetaiotaomicron, and to identify the genes involved to enable investigation of potential mechanisms for the generation of metronidazole resistance. Methods: The genes affected by the transposon insertion were identified by plasmid rescue and sequencing. Expression levels of the relevant genes were determined by semi-quantitative RNA hybridization and catabolic activity by lactate dehydrogenase/pyruvate oxidoreductase assays. Results: A metronidazole-resistant mutant was isolated and the transposon insertion site was identified in an intergenic region between the rhaO and rhaR genes of the gene cluster involved in the uptake and catabolism of rhamnose. Metronidazole resistance was observed during growth in defined medium containing either rhamnose or glucose. The metronidazole-resistant mutant showed improved growth in the presence of rhamnose as compared with the wild-type parent. There was increased transcription of all genes of the rhamnose gene cluster in the presence of rhamnose and glucose, likely due to the transposon providing an additional promoter for the rhaR gene, encoding the positive transcriptional regulator of the rhamnose operon. The B. thetaiotaomicron metronidazole resistance phenotype was recreated by overexpressing the rhaR gene in the B. thetaiotaomicron wild-type parent. Both the metronidazole-resistant transposon mutant and RhaR overexpression strains displayed a phenotype of higher lactate dehydrogenase and lower pyruvate oxidoreductase activity in comparison with the parent strain during growth in rhamnose. Conclusions: These data indicate that overexpression of the rhaR gene generates metronidazole resistance in B. thetaiotaomicron

Natural Abs, which arise without known immune exposure, have been described that specifically recognize cells dying from apoptosis, but their role in innate immunity remains poorly understood. Herein, we show that the immune response to neoantigenic determinants on apoptotic thymocytes is dominated by Abs to oxidation-associated Ags, phosphorylcholine (PC), a head group that becomes exposed during programmed cell death, and malondialdehyde (MDA), a reactive aldehyde degradation product of polyunsaturated lipids produced following exposure to reactive oxidation species. While natural Abs to apoptotic cells in naive adult mice were dominated by PC and MDA specificities, the amounts of these Abs were substantially boosted by treatment of mice with apoptotic cells. Moreover, the relative amounts of PC and MDA Abs was affected by VH gene inheritance. Ab interactions with apoptotic cells also mediated the recruitment of C1q, which enhanced apoptotic cell phagocytosis by immature dendritic cells. Significantly, IgM Abs to both PC and MDA were primary factors in determining the efficiency of serum-dependent apoptotic cell phagocytosis. Hence, we demonstrate a mechanism by which certain natural Abs that recognize neoantigens on apoptotic cells, in naive mice and those induced by immune exposure to apoptotic cells, can enhance the functional capabilities of immature dendritic cells for phagocytic engulfment of apoptotic cells.

Bacteroides thetaiotaomicron is an important human gut commensal, which also causes opportunistic infections outside this environment. It utilises a range of host and diet-related carbohydrates, including rhamnose. In this study, the rha gene cluster, required for rhamnose utilisation, was characterised by transcription analysis, gene targeted mutagenesis and enzyme assays. Growth in the presence of L-rhamnose induced transcription of all the genes of this cluster. The first five genes of the cluster, rhaKIPAO, were transcribed as an operon from a transcriptional start site upstream of rhaK, whereas the sixth gene, rhaR, was transcribed independently. Bioinformatic analysis and mutation of the rhaR gene identified it as encoding the positive transcriptional activator of rhaKIPAO. A rhaR mutant could not utilise rhamnose as the sole carbon source but grew normally on glucose. The rhaO gene encoded a lactaldehyde reductase, and a rhaO mutant produced reduced levels of L-1,2-propanediol during growth in rhamnose, indicating its contribution to rhamnose catabolism in Bacteroides thetaiotaomicron.

Prehospital personnel in Emergency Medical Service (EMS) systems have varying levels of accuracy in stroke recognition. Identifying the accuracy of emergency medical dispatcher using Medical Priority Dispatch Systems (MPDS) stroke protocol and paramedics may help understand the accuracy of stroke recognition in about 3000 emergency medical dispatch systems and prehospital systems world wide.Our aim was to assess the accuracy of stroke identification in emergency medical dispatchers (EMD) with high compliance to MPDS protocol and paramedics using Cincinnati Prehospital Stroke Scale (CSS).This was a retrospective observational study. Data was acquired from a computer assisted dispatch (CAD) system, a computerized paramedic record database and discharge diagnosis from billing records or stroke registry containing all stroke assessments of patients who presented to the participating study hospitals within 12 hours of symptom onset. We included patients 18 years or older, identified as having stroke by EMD and city agency paramedics. We excluded patients taken to hospitals not participating in the study, patients with a dispatch determinant of Stroke (card 28) not transported by City EMS agency (SDMSE) to participating hospitals, patients in the stroke registry not transported by SDMSE or patients with no final outcome data. A stroke neurologist or hospital discharge diagnosis of stroke (physician diagnosis) was used to determine the sensitivity and predictive values of EMD and paramedic recognition of stroke.Of 882 patients with a dispatch determinant of stroke using MPDS Stroke protocol, 367 had a final discharge diagnosis of stroke. This gives a sensitivity of 83% and a positive predictive value of 42% for EMD using MPDS Stroke protocol. Of 477 patients with a paramedic assessment of stroke using CSS, 193 had a final discharge diagnosis of stroke. This gives a sensitivity of 44% and a PPV of 40% for paramedics using CSS.In our EMS system, EMD using MPDS Stroke protocol with a high compliance has a higher sensitivity than paramedics using CSS.

Graphical abstract, Highlights • Genotyping and gene sequencing reveal limited diversity in a live Theileria parva vaccine. • Two of the vaccine components show a high level of similarity at all loci. • The vaccine contains very little of the diversity found in field T. parva. • The vaccine is suboptimal for generating immunity again diverse parasite strains. • The presence of minor genotypic components poses difficulties for quality control., An infection and treatment protocol is used to vaccinate cattle against Theileria parva infection. Due to incomplete cross-protection between different parasite isolates, a mixture of three isolates, termed the Muguga cocktail, is used for vaccination. While vaccination of cattle in some regions provides high levels of protection, some animals are not protected against challenge with buffalo-derived T. parva. Knowledge of the genetic composition of the Muguga cocktail vaccine is required to understand how vaccination is able to protect against field challenge and to identify the potential limitations of the vaccine. The aim of the current study was to determine the extent of genetic and antigenic diversity within the parasite isolates that constitute the Muguga cocktail. High throughput multi-locus sequencing of antigen-encoding loci was performed in parallel with typing using a panel of micro- and mini-satellite loci. The former focused on genes encoding CD8+ T cell antigens, believed to be relevant to protective immunity. The results demonstrate that each of the three component stocks of the cocktail contains limited parasite genotypic diversity, with single alleles detected at many gene/satellite loci and, moreover, that two of the components show a very high level of similarity. Thus, the vaccine incorporates very little of the genetic and antigenic diversity observed in field populations of T. parva. The presence of alleles at low frequency (

The Infection and Treatment Method (ITM) of vaccination against the apicomplexan parasite Theileria parva has been used since the early 1970s and is still the only commercially available vaccine to combat the fatal bovine disease, East Coast fever (ECF). The disease is tick-transmitted and results in annual economic losses of at least $300 million per year. While this vaccine technology has been available for over 40 years, few attempts have been made to standardize the production process and characterize the vaccine. The latest batch was produced in early 2008 at the International Livestock Research Institute (ILRI). The vaccine production involves the use of cattle free from parasites routinely monitored throughout the production process, and a pathogen-free tick colony. This paper describes the protocol used in the recent production, and the process improvements, including improved quality control tools, that had not been employed in previous ITM productions. The paper also describes the processes involved in determining the appropriate field dose, which involved a three-step in vivo study with various dilutions of the vaccine stabilate. The vaccine was shown to be safe and viable after production, and a suitable field dose was identified as 1 ml of a 1:100 dilution.

Background Combination antiretroviral therapy (cART) has resulted in a dramatic decrease in human immunodeficiency virus (HIV)-related opportunistic infections and deaths in US youth, but both continue to occur. Methods We estimated the incidence of complications and deaths in IMPAACT P1074, a long-term US-based prospective multicenter cohort study conducted from April 2008 to June 2014. Incidence rates of selected diagnoses and trends over time were compared with those from a previous observational cohort study, P219C (2004-2007). Causes of death and relevant demographic and clinical features were reviewed. Results Among 1201 HIV-infected youth in P1074 (87% perinatally infected; mean [standard deviation] age at last chart review, 20.9 [5.4] years), psychiatric and neurodevelopmental disorders, asthma, pneumonia, and genital tract infections were among the most common comorbid conditions. Compared with findings in P219C, conditions with significantly increased incidence included substance or alcohol abuse, latent tuberculosis, diabetes mellitus, atypical mycobacterial infections, vitamin D deficiency or metabolic bone disorders, anxiety disorders, and fractures; the incidence of pneumonia decreased significantly. Twenty-eight deaths occurred, yielding a standardized mortality rate 31.5 times that of the US population. Those who died were older, less likely to be receiving cART, and had lower CD4 cell counts and higher viral loads. Most deaths (86%) were due to HIV-related medical conditions. Conclusions Opportunistic infections and deaths are less common among HIV-infected youth in the US in the cART era, but the mortality rate remains elevated. Deaths were associated with poor HIV control and older age. Emerging complications, such as psychiatric, inflammatory, metabolic, and genital tract diseases, need to be addressed.

Background Heart failure is common in the elderly and is associated with high rates of hospitalisation, readmission and mortality. International guidelines however are not frequently implemented in this population. Methods We retrospectively studied the clinical profile, investigations, treatment on discharge, length of hospital stay, readmission rate and mortality in 261 patients, aged ≥75 years, with a discharge diagnosis of heart failure. Clinical frailty was estimated using the Canadian Study of Health and Aging clinical frailty scale. Results Hypertension (64%), atrial fibrillation (50.6%) and ischaemic heart disease (46%) were common, and 75.6% of patients were clinically vulnerable or frail. 23.5% of admitters had an inpatient echocardiogram and 20% of patients had at least one readmission episode for heart failure. On discharge, 64.6% of admissions were treated with an ACE inhibitor or angiotensin II receptor antagonist, 49.3% with a β blocker and 28.7% with an aldosterone receptor antagonist (ARA). Patients discharged from cardiology wards were more likely to receive a β blocker (p

Objective We studied the complications and timing implications of prehospital activated charcoal (PAC). Appropriateness of PAC administration was also evaluated. Methods We retrospectively reviewed prehospital records over 32 months for overdose cases, where PAC was administered. Cases were assessed for amount and type of ingestant, clinical findings, timing of PAC, timing of transport and arrival into the emergency department (ED), and complications. Encounter duration in cases of PAC was compared with that, for all cases during the study period, where an overdose patient who did not receive activated charcoal was transported. Results Two thousand eight hundred forty-five total cases were identified. In 441 cases, PAC was given; and complications could be assessed. Two hundred eighty-one of these had complete information regarding timing of ingestion, activated charcoal administration, and transport. The average time between overdose and PAC was 49.8 minutes (range, 7-199 minutes; median, 41.0 minutes; SD, 30.4 minutes). Complications included emesis (7%), declining mental status (4%), declining blood pressure (0.4%), and declining oxygen saturation (0.4%). Four hundred seventeen cases of PAC had documentation of timing of emergency medical service (EMS) arrival on scene and arrival at the ED. Average EMS encounter time was 29 minutes (range, 10-53 minutes; median, 27.9 minutes). Two thousand forty-four poisoning patients were transported who did not receive PAC. The average EMS encounter time for this group was 28.1 minutes (range, 4-82 minutes; median, 27.3 minutes), not significantly different ( P =.114). Conclusions Prehospital activated charcoal did not appear to markedly delay transport or arrival of overdose patients into the ED and was generally safe.

Crohn9s disease (CD) has a bimodal distribution in incidence, with a second peak in the elderly. However, its diagnosis in the elderly is difficult due to a wider range of more common differential diagnoses such as diverticulitis, ischaemic colitis and colorectal cancer. We report a likely case of CD in a 95-year-old woman. She presented with diarrhoea and rectal bleeding and was found to have multiple pleomorphic ulcers with a patchy cobblestone mucosa on sigmoidoscopy. Histopathology demonstrated focal ulceration, altered crypt architecture and adjacent neutrophil polymorph infiltration with no granolomata or features of malignancy. The patient passed away after steroid treatment was started. This case is a reminder that CD can present in the elderly and highlights the challenging diagnosis and high mortality of CD-related hospitalisation in the elderly. When considering management, attention should be given to comorbid disease, age-related changes in pharmacokinetics and patients social circumstances.

Background The role of IL-7 and pre-TCR signaling during T cell development has been well characterized in murine but not in human system. We and others have reported that human BM hematopoietic progenitor cells (HPCs) display poor proliferation, inefficient double negative (DN) to double positive (DP) transition and no functional maturation in the in vitro OP9-Delta-like 1 (DL1) culture system. Results In this study, we investigated the importance of optimal IL-7 and pre-TCR signaling during adult human T cell development. Using a modified OP9-DL1 culture ectopically expressing IL-7 and Fms-like tyrosine kinase 3 ligand (Flt3L), we demonstrated enhanced T cell precursor expansion. IL-7 removal at various time points during T cell development promoted a slight increase of DP cells; however, these cells did not differentiate further and underwent cell death. As pre-TCR signaling rescues DN cells from programmed cell death, we treated the culture with anti-CD3 antibody. Upon pre-TCR stimulation, the IL-7 deprived T precursors differentiated into CD3+TCRαβ+DP cells and further matured into functional CD4 T cells, albeit displayed a skewed TCR Vβ repertoire. Conclusions Our study establishes for the first time a critical control for differentiation and maturation of adult human T cells from HPCs by concomitant regulation of IL-7 and pre-TCR signaling.

The role of IL-7 in pre-T cell receptor (TCR) signaling during human T cell development is poorly understood. To study this, we engineered Molt3, a T cell progenitor T-acute lymphoblastic leukemia cell line, using lentiviral IL-7 receptor α (IL-7Rα) to serve as a model system. IL-7 promoted pre-TCR activation in IL-7Rα(hi) Molt3 as illustrated by CD25 up-regulation after anti-CD3 stimulation. Anti-CD3 treatment activated Akt and Erk1/2 signaling pathways as proven using specific inhibitors, and IL-7 further enhanced both signaling pathways. The close association of IL-7Rα with CD3ζ in the pre-TCR complex was illustrated through live imaging confocal fluorescence microscopy. These results demonstrate a direct and cooperative role of IL-7 in pre-TCR signaling.

A small group of adults disproportionately and ineffectively use acute services including emergency medical services (EMS) and emergency departments (EDs). The resulting episodic, uncoordinated care is of lower quality and higher cost and simultaneously consumes valuable public safety and acute care resources.To address this issue, we measured the impact of a pilot, EMS-based case management and referral intervention termed the San Diego Resource Access Program (RAP) to reduce EMS, ED, and inpatient (IP) visits.This was a historical cohort study of RAP records and billing data of EMS and one urban hospital for 51 individuals sequentially enrolled in the program. The study sample consisted of adults with ≥ 10 EMS transports within 12 months and others reported by prehospital personnel with significant recent increases in transports. Data were collected over a 31-month time period from December 2006 to June 2009. Data were collected for equal pre- and postenrollment time periods based on date of initial RAP contact, and comparisons were made using the Wilcoxon signed-rank test. Overall use for subjects is reported.The majority of subjects were male (64.7%), homeless (58.8%), and 40 to 59 years of age (72.5%). Between the pre and post periods, EMS encounters declined 37.6% from 736 to 459 (p = 0.001), resulting in a 32.1% decrease in EMS charges from $689,743 to $468,394 (p = 0.004). The EMS task time and mileage decreased by 39.8% and 47.5%, respectively, accounting for 262 (p = 0.008) hours and 1,940 (p = 0.006) miles. The number of ED encounters at the one participating hospital declined 28.1% from 199 to 143, which correlated with a 12.7% decrease in charges from $413,410 to $360,779. The number of IP admissions declined by 9.1% from 33 to 30, corresponding to a 5.9% decrease in IP charges from $687,306 to $646,881. Hospital length of stay declined 27.9%, from 122 to 88 days. Across all services, total charges declined by $314,406.This pilot study demonstrated that an EMS-based case management and referral program was an effective means of decreasing EMS transports by frequent users, but had only a limited impact on use of hospital services.

Background The use of continuous positive airway pressure (CPAP)-assisted ventilation in the prehospital setting has not been well studied. Objectives The purpose of this study was to measure the efficacy of adding prehospital CPAP to an urban emergency medical services (EMS) respiratory distress protocol for persons with respiratory distress. Methods An historical cohort analysis of consecutive EMS patients presenting during the years 2005–2010. Groups were matched for severity of respiratory distress. Physiologic variables were the primary outcomes obtained from first responders and upon triage in the Emergency Department. Additional outcomes included endotracheal intubation rate, hospital mortality, overall hospital length of stay (LOS), intensive care unit (ICU) admission, and ICU LOS. Results There were a total of 410 consecutive patients with predetermined criteria for severe respiratory distress, 235 historical controls matched with 175 post-implementation patients, entered in the study. The average age was 67 years; 54% were men. There were significant median differences in heart and respiratory rates favoring the historical cohort (all p 0.05). Patients who were continued on non-invasive ventilatory assistance had a significantly improved rate of intubation and ICU LOS (all p Conclusion The addition of CPAP to an EMS prehospital respiratory distress protocol resulted in improved heart and respiratory rates. Though not statistically significant, decrease in overall and ICU LOS were observed. Patients with continued ventilatory assistance seemed to have improved rates of intubation and ICU LOS.

Despite recent improvements to in situ proteolysis strategies, a higher efficiency is still needed to increase both the number of peptides detected and the associated ion intensity, leading to a complete and reliable set of biomarkers for diagnostic or prognostic purposes. In the study presented here, an extract of a systematic study is illustrated investigating a range of surfactants assisting trypsin proteolytic activity. Method development was trialled on fingermarks; this specimen results from a transfer of sweat from an individual's fingertip to a surface upon contact. As sweat carries a plethora of biomolecules, including peptides and proteins, fingermarks are, potentially, a very valuable specimen for non-invasive prognostic or diagnostic screening. A recent study has demonstrated the opportunity to quickly detect peptides and small proteins in fingermarks using Matrix Assisted Laser Desorption Ionization Mass Spectrometry Profiling (MALDI MSP). However, intact detection bears low sensitivity and does not allow species identification; therefore, a shotgun proteomic approach was employed involving in situ proteolysis. Data demonstrate that in fingermarks, further improvements to the existing method can be achieved using MEGA-8 as surfactant in higher percentages as well as combinations of different detergents. Also, for the first time, Rapigest SF, normally used in solution digestions, has been shown to successfully work also for in situ proteolysis.

Recent progress in dendritic cell (DC) and T cell therapies has offered a new avenue in treating multiple myeloma (MM). We have reported that MM patients' DCs cannot effectively present MM antigens and, instead, preferrentially activate T regulatory cells (Tregs). Furthermore, the MM cells actively suppress patients' DC and T cell functions (Han et al. 2008 Mol Ther. 16, 269–279). Therefore, an effective immunotherapy must overcome immune tolerance to the cancer cells and effectively expand functional MM-specific effector cells. To obtain critical data about MM patient's immune status, a longitudinal study of Treg cells before and after treatments of twenty MM patients was conducted. Patients' bone marrow (BM) and peripheral blood (PB) specimens were collected in the first visit and the follow up visits during treatment. Treg cell populations were examined using antibodies against CD4, CD25, CD127 and Foxp3. At the same time, naïve versus memory phenotype was assessed using antibodies against CD45RA, CCR7 and CD27. Besides the above markers, the surface molecule KLRG1 reflecting T cell senescence and longivity was also detected by flow cytometry. Analysis of Tregs at different time points showed a clear trend of gradual increase in Treg population both in the BM and PB of the MM patients; compared with healthy donors, MM patients' Tregs expand more than two to three folds. Surface marker analysis suggests that the expanded Tregs are mostly memory effector cells, which coincides with a decrease in the ratio of naïve Tregs. Analysis of MM-specific T cell responses by examining effector cytokines and cytolytic activities against autologous tumor cells revealed that BM contained more cytotoxic T cells than did PB. Increased expansion of MM-specific immune effector cells in culture was observed when Treg activities were suppressed using various immune modulators. Further understanding of Treg-mediated cancer tolerance will be essential to develop clinically more effective immunotherapy towards improved survival of MM patients. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-157. doi:10.1158/1538-7445.AM2011-LB-157