Evaluation of digital staining for ex vivo confocal laser scanning microscopy.

BACKGROUND: Ex vivo confocal laser scanning microscopy (CLSM) is a novel diagnostic tool for the fast examination of native tissue. However, CLSM produces black/white/green images, depending on the refraction indices of the tissue structures, complemented by nuclear fluorescence staining, which the vast majority of Mohs surgeons and dermatopathologists are not trained to interpret. Digital staining is applicable to ex vivo CLSM investigations to simulate the images of conventional slides stained with haematoxylin and eosin (H&E). OBJECTIVES: The aim of our study was to evaluate in detail the appearance of human skin structures using digitally stained ex vivo CLSM images and compare the results to that of conventional H&E slides of the same specimen. METHODS: After providing informed consent, 26 patients donated their Burow's triangles (healthy skin) that resulted from plastic reconstruction after the R0 excision of skin tumours. After being investigated by ex vivo CLSM, including automated digital staining (VivaScope 2500M-4G, MAVIG GmbH), the specimens were fixed in formalin, embedded in paraffin and stained with H&E. RESULTS: Almost all skin structures in the digitally stained ex vivo CLSM images morphologically resembled the structures in the histopathological images acquired from H&E slides. Due to the high refraction index of melanin, the hair shafts appeared bright pink, and the melanocytes and melanophages were poorly imaged, resulting in a strong pink appearance that vastly differed from the appearance of conventional H&E-stained histopathology. CONCLUSIONS: Digital staining of ex vivo CLSM images is an easy and highly useful tool to facilitate the interpretation of black-field images generated by confocal laser scanning microscopy for dermatopathologists and Mohs surgeons who are familiar with H&E staining. Unlike the pigmented structures, the cutaneous and subcutaneous structures had excellent visualization with only minimal differences from their appearance on H&E slides.

Morbidity and mortality of candidaemia in Europe: an epidemiologic meta-analysis.

OBJECTIVES: Candidaemia is a serious hazard to hospitalized patients, but European epidemiological data are restricted to national studies focusing on Northern Europe, population-based surveillance programmes or studies conducted in distinct local areas. The aim was to provide current data on the overall burden and epidemiological development of candidaemia in Europe. METHODS: A Web of Knowledge™ search was carried out from January 2000 to February 2019. Appropriate data were collected on total cases, study duration, incidence, species distribution and/or mortality rates. Meta-analysis was performed to pool individual studies. Heterogeneity was examined using the I2 statistic. Calculations of pooled incidence and mortality rates, subgroup analysis by geographical origin, study period and scenarios were carried out. Daily candidaemia incidence and mortality rates in Europe were extrapolated. Systematic review and meta-analysis were used to determine incidence and mortality of candidaemia in the UN European region. Complete datasets were categorized into population-based and hospital-based epidemiological studies and were analysed separately. Subgroup analyses were performed for geographic distributions and time-dependent developments. RESULTS: In population-based studies, 43 799 cases of candidaemia were diagnosed in 1 885 271 885 person-years, revealing an overall pooled incidence rate of 3.88/100 000. The highest pooled incidence rate was observed in intensive care units (5.5/1000 admissions, Day 30 mortality rate 37%), followed by tertiary care centres (0.96/1000 admissions, pooled Day 30 mortality rate 38%) and the mixed group of teaching and general hospitals (0.52/1000 admissions, pooled Day 30 mortality rate 37%). European incidence of candidaemia was extrapolated to approximately 79 cases per day, of which an estimated 29 patients might have fatal outcome at Day 30. CONCLUSIONS: Pooled incidence rates, species distribution and outcome of candidaemia differ considerably between clinical groups, European regions and over time. We observed an increasing overall pooled incidence rate of candidaemia and a higher proportion of Candida spp. other than C. albicans in the current decade in population-based data.

Impact of single-room contact precautions on hospital-acquisition and transmission of multidrug-resistant Escherichia coli: a prospective multicentre cohort study in haematological and oncological wards.

OBJECTIVES: Colonization and infection with third-generation cephalosporin-resistant Escherichia coli (3GCR-EC) are frequent in haematological and oncological patients. In this high-risk setting, German guidelines recommend single-room contact precautions (SCP) for patients with 3GCR-EC that are non-susceptible to fluoroquinolones (F3GCR-EC). However, this recommendation is controversial, as evidence is limited. METHODS: We performed a prospective, multicentre cohort study at four haematology and oncology departments assessing the impact of SCP on hospital-acquired colonization or bloodstream infection (BSI) with F3GCR-EC. Two sites performed SCP for F3GCR-EC patients including single rooms, gloves and gowns (SCP sites), and two did not (NCP sites). Active screening for 3GCR-EC was performed and isolates were characterized with molecular typing methods including whole genome sequencing and core genome multiple locus sequence typing to assess patient-to-patient transmission. Potential confounders were assessed by competing-risk regression analysis. RESULTS: Within 12 months, 1386 patients at NCP sites and 1582 patients at SCP sites were included. Hospital-acquisition of F3GCR-EC was observed in 22/1386 (1.59%) and 16/1582 (1.01%) patients, respectively (p 0.191). There were 3/1386 (0.22%) patients with BSI caused by F3GCR-EC at NCP sites and 4/1582 (0.25%) at SCP sites (p 1.000). Patient-to-patient transmission occurred in three cases at NCP and SCP sites each (p 1.000). The number of patients needed to screen in order to prevent one patient-to-patient transmission of F3GCR-EC was determined to be 3729. CONCLUSIONS: Use of SCP had no significant impact on hospital-acquisition or patient-to-patient transmission of F3GCR-EC in this high-risk setting.

Pretreatment human immunodeficiency virus type 1 (HIV-1) drug resistance in transmission clusters of the Cologne-Bonn region, Germany.

OBJECTIVES: In Germany, previous reports have demonstrated transmitted human immunodeficiency virus type 1 (HIV-1) drug-resistance mutations (DRM) in 11% of newly diagnosed individuals, highlighting the importance of drug-resistance screening before the initiation of antiretroviral therapy (ART). Here, we sought to understand the molecular epidemiology of HIV DRM transmission in the Cologne-Bonn region of Germany, given one of the highest rates of new HIV diagnoses in western Europe (13.7 per 100 000 habitants). METHODS: We analysed 714 HIV-1 ART-naive infected individuals diagnosed at the University Hospitals Cologne and Bonn between 2001 and 2016. Screening for DRM was performed according to the Stanford University Genotypic Resistance Interpretation. Shared DRM were defined as any DRM present in genetically linked individuals (<1.5% genetic distance). Phylogenetic and network analyses were performed to infer putative relationships and shared DRM. RESULTS: The prevalence of any DRM at time of diagnosis was 17.2% (123/714 participants). Genetic transmission network analyses showed comparable frequencies of DRM in clustering versus non-clustering individuals (17.1% (85/497) versus 17.5% (38/217)). The observed rate of DRM in the region was higher than previous reports 10.8% (87/809) (p < 0.001), revealing the need to reduce onward transmission in this area. Genetically linked individuals harbouring shared DRM were more likely to live in suburban areas (24/38) than in central Cologne (1/38) (p < 0.001). CONCLUSION: The rate of DRM was exceptionally high. Network analysis elucidated frequent cases of shared DRM among genetically linked individuals, revealing the potential spread of DRM and the need to prevent onward transmission of DRM in the Cologne-Bonn area.

Measles, mumps, rubella and VZV: importance of serological testing of vaccine-preventable diseases in young adults living with HIV in Germany.

Measles, mumps, rubella (MMR) and varicella zoster virus (VZV) infection can cause serious diseases and complications in the HIV-positive population. Due to successful vaccination programmes measles, mumps and congenital rubella syndrome has become neglected in Germany. However, recent outbreaks of measles have occurred from import-associated cases. In this cross-sectional study the serostatus for MMR and VZV in 2013 HIV-positive adults from three different university outpatient clinics in Bonn (n = 544), Cologne (n = 995) and Munich (n = 474) was analysed. Sera were tested for MMR- and VZV-specific immunglobulin G antibodies using commercial immunoassays. Seronegativity was found in 3% for measles, 26% for mumps, 11% for rubella and 2% for VZV. Regarding MMR, 35% of patients lacked seropositivity against at least one infectious agent. In multivariable analysis younger age was strongly associated with seronegativity against all four viruses, measles, mumps, rubella (P < 0·001, P < 0·001 and P = 0·001, respectively) and VZV (P = 0·001). In conclusion, there is high need for MMR and VZV vaccination in people living with HIV in Germany born in 1970 or later. Thus, systematic MMR and VZV antibody screening and vaccination should be implemented in the HIV-positive population to prevent serious disease and complications of vaccine-preventable diseases.

Prematuros tardíos: estudio clínico retrospectivo de sus causas y consecuencias / Late preterm infants: a retrospective clinical study of their causes and consequences

ANTECEDENTES: Los niños nacidos entre las 34 0/7 y 36 6/7 semanas se denominan prematuros tardíos (PT), constituyen 5-7% de los nacidos y poseen mayor morbimortalidad que los niños de término. OBJETIVO: Analizar las causas de partos PT en nuestra institución. Comparar morbilidad neonatal de nacidos PT y de término. MÉTODOS: Estudio de cohorte retrospectivo. Se revisan registros de nacimientos entre enero de 2009 y diciembre de 2012 identificando los nacidos vivos entre las 34 0/7 y 36 6/7 y entre las 39 0/6 y 40 6/7 semanas. Se identifica grupo clínico de parto prematuro al que pertenecen. Se compara frecuencia de resultados perinatales de nacidos PT y de término completo. RESULTADOS: En el período estudiado nacieron 8.890 niños vivos. Tasa de partos PT fue 7,49% (n = 666). El 64,11% fue por causa idiopática o asociado a rotura de membranas y 35,89% por indicación médica. Dentro del grupo de indicaciones médicas un 19,25% de interrupciones no están basadas en evidencia y ninguna puramente electiva. Los PT, comparados con niños de término, tienen más riesgo de hospitalización y morbilidad neonatal; este riesgo es mayor a menor edad gestacional. CONCLUSIONES: En nuestro hospital 1 de cada 5 PT pudo haber nacido a una edad gestacional mayor, lo que podría haber evitado los riesgos perinatales asociados. Se sugiere que las instituciones de salud analicen las causas de interrupciones del embarazo en el grupo de PT, lo que podría ser un indicador de calidad obstétrica.

BACKGROUND: Children born between 34 0/7 and 36 6/7 weeks of gestation are called late-preterm infants. Represent 5 to 7% of live births and they are at higher risk of morbidity and mortality than term infants. OBJECTIVE: To analyze causes of late preterm births in our institution. To compare neonatal morbidity in late-preterm and term infants. METHODS: A retrospective cohort study. Birth records between January 2009 and December 2012 were reviewed identifying live births between 34 0/7 and 36 6/7 and between 39 0/6 and 40 6/7 weeks of gestation. Clinical group of preterm labor to which they belong was identified. Frequency of perinatal outcomes of late-preterm and term infants were compared. RESULTS: In the period under study there were 8890 live birth. The late-preterm birth rate was 7.49% (n = 666); 64.11% of them was idiopathic in cause or associated with rupture of membranes and 35.89% was for medical indication. In the group of medical indications 19.25% of the interruptions were not based on evidence and none were purely elective. Late-preterm infants have higher risk of hospitalization and neonatal morbidity than term infants; this risk is higher at earlier gestational ages. CONCLUSIONS: In our hospital 1 in 5 late-preterm birth could have been born at higher gestational age, preventing perinatal risks. It is suggested that health institutions analyze causes of interruption of pregnancy in the late-preterm group, which could constitute an indicator of obstetric quality.

Effects of umbrella palms and wastewater depth on wastewater treatment in a subsurface flow constructed wetland.

On-site subsurface flow constructed wetlands are designed to provide secondary quality effluent. Plants and wetland volume are considered in their design. There have been no studies, however, comparing wastewater treatment at different wastewater depths, and plant effects in wetlands are not completely understood. Investigations were conducted on these variables using four wetland cells 228 m wide by 4.75 m long containing 1 to 5 cm diameter river rock. Cyperus alternifolius (umbrella palms) were planted in one cell, and side-by-side comparisons were made between the planted and a control cell. Side-by-side comparisons were also made between cells with equal surface areas and different depths. At best umbrella palms improved effluent 5-day biochemical oxygen demand (BOD5) by 8%, suspended solids by 6%, and did not improve fecal coliform or P wastewater quality in July. When ambient air temperatures were < or = 12 degrees C during December, plants did not improve most treatment parameters. They did, however, significantly improve NH4+ treatment even when ambient air temperatures were as low as 8 degrees C. Increasing wastewater depth enhanced fecal coliform die-off but did not reduce effluent considered when maximum NH4+ reduction is a BOD5, suspended solids, NH4+, or P in effluent. Umbrella palms should be treatment goal and it is not necessary to reduce other parameters. Surface area was more important to wastewater treatment than depth.

Water flow patterns in subsurface flow constructed wetlands designed for on-site domestic wastewater treatment.

Bypass flow in subsurface flow constructed wetlands, which may be related to several wetland characteristics, reduces detention time and may result in inadequately treated wastewater. Subsurface flow constructed wetlands, 2.3 m wide by 4.8 m long and containing a gravel matrix, were used to investigate the impact of wetland depth, inlet pipe location, loading volume, and plants on water flow. Flow patterns were determined using blue dye or bromide as tracers. The blue dye adsorbed to the gravel and was not an effective tracer for following water movement. Water dispersed as it flowed through the wetland, and approximately two pore volumes of added water were required to displace approximately 99% of the bromide tracer added as a pulse. In 17 and 25 cm deep wetlands, water flow was uniform with depth, and inlet depth had little influence on water flow patterns. Water flow in a 40 cm deep wetland was not uniform with depth for either inlet pipe placement. The presence of plants caused preferential water flow around root masses, thereby, limiting their potential to interface with wastewater. Water mixing by dispersion from the surface to deeper depths may enhance aeration. Bromide was first detected in effluent after only 0.5 pore volumes of tap water had been added. This indicates that detention time for some wastewater would be less than expected, since plug flow is usually assumed in subsurface flow constructed wetlands. This occurrence should be considered for time-dependent treatments, such as fecal coliform and biological oxygen demand reduction.