Cytokine Release Assays as Tests for Exposure to Leishmania,and for Confirming Cure from Leishmaniasis,in Solid Organ Transplant Recipients

Spain has one of the world’s largest pools of organ donors and is a global leader in terms of the number of transplants it performs. The current outbreak of leishmaniasis in Fuenlabrada (in the southwest of the region of Madrid, Spain) has involved 600 clinical cases since late 2009 (prevalence 0.2%). It may therefore be wise to monitor the town’s transplanted population for Leishmania infantum; its members are immunosuppressed and at greater risk of infection and relapse following treatment. The present work examines the use of cytokine release assays to determine the prevalence of Leishmania infection in this population, and to confirm recovery following treatment for visceral leishmaniasis (VL). The humoral and cellular immune responses to L. infantum were characterized in 63 solid organ transplant (SOT) recipients from Fuenlabrada, 57 of whom reported no previous episode of VL (NVL subjects), and six of whom had been cured of VL (CVL subjects). Seventeen subjects (12 NVL and 5 CVL) showed a patent lymphoproliferative response to soluble Leishmania antigen (SLA). Stimulation of peripheral blood mononuclear cell cultures and of whole blood with SLA led to the production of different combinations of cytokines that might serve to confirm Leishmania infection or recovery from VL and help prevent cured patients from relapsing into this serious condition.     

Rind Structure, Epicuticular Wax Morphology and Water Permeability of 'Fortune' Mandarin Fruits affected by Peel Pitting

Rind structure and surface wax morphology of 'Fortune' mandarinsaffected by pre-harvest peel pitting were examined as relatedto the severity of this disorder. In fruits with slight symptomsof pitting (< 10% surface area pitted) a discrete numberof epidermal cells were injured with signs of cellular breakdown.In fruits with moderate symptoms of pitting (10-50% surfacearea pitted) the injured zones also involved hypodermal cells.The surface of rind had an undulating appearance in fruits withsevere symptoms of pitting (> 50% surface area pitted). Thedepressed areas in these fruits were devoid of crystalline waxstructures and had a rough morphology. Broad regions of rindhad crushed epidermal and hypodermal cells with infolded walls.These cells were either empty or filled with reduced cytoplasmamounts located in central positions of the cell. No signs ofdisruption in the cuticle were observed. Under field conditions,pitting occurrence coincided with higher permeability valuesof isolated cuticles. Furthermore, length of cold treatmentsrequired to show post-harvest pitting symptoms was shorter infruits sampled at this stage than in those collected earlieror later. Our results indicate that the low temperatures inducethe breakdown of the more external cellular strata of the rind.Thus, the physiological function of cuticle is highly impairedwhich, in turn, increases water permeability considerably.Copyright1994, 1999 Academic Press Chilling injury, Citrus, cold storage, pre-harvest pitting     

<Emphasis Type="Italic">Membraniporopsis tubigera</Emphasis>, an invasive bryozoan in sandy beaches of southern Brazil and Uruguay

The massive irruption of the invasive bryozoan Membraniporopsis tubigera (Osburn) in sandy beaches of southern Brazil and Uruguay is reported. The species, originally described from Puerto Rico, Texas and Florida, has also been recorded for Brazilian beaches from 21°S to 26°S as well as for harbours of Australia, New Zealand and the Sea of Japan. The southward spreading rate of this bryozoan along the Brazilian and Uruguayan coasts can be estimated in approximately 183–195 km year⁻¹. The chances that this invasion could proceed southwards in the Southwest Atlantic and the possible impacts that it may be causing are discussed. The case of M. tubigera seems to be qualitatively and quantitatively different from those of other alien bryozoans previously recorded for this region, since it appeared massively in exposed sandy beaches, a habitat regarded to date as apparently free from the pervasive ecological impact of invasion by exotic species in the Southwest Atlantic.     

The Prognostic Value of BRCA1 mRNA Expression Levels Following Neoadjuvant Chemotherapy in Breast Cancer

Background

A fraction of sporadic breast cancers has low BRCA1 expression. BRCA1 mutation carriers are more likely to achieve a pathological complete response with DNA-damage-based chemotherapy compared to non-mutation carriers. Furthermore, sporadic ovarian cancer patients with low levels of BRCA1 mRNA have longer survival following platinum-based chemotherapy than patients with high levels of BRCA1 mRNA.

Methodology/Principal Findings

Tumor biopsies were obtained from 86 breast cancer patients who were candidates for neoadjuvant chemotherapy, treated with four cycles of neoadjuvant fluorouracil, epirubicin and cyclophosphamide. Estrogen receptor (ER), progesterone receptor (PR), HER2, cytokeratin 5/6 and vimentin were examined by tissue microarray. HER2 were also assessed by chromogenic in situ hybridization, and BRCA1 mRNA was analyzed in a subset of 41 patients for whom sufficient tumor tissue was available by real-time quantitative PCR. Median time to progression was 42 months and overall survival was 55 months. In the multivariate analysis for time to progression and overall survival for 41 patients in whom BRCA1 could be assessed, low levels of BRCA1 mRNA, positive PR and negative lymph node involvement predicted a significantly lower risk of relapse, low levels of BRCA1 mRNA and positive PR were the only variables associated with significantly longer survival.

Conclusions/Significance

We provide evidence for a major role for BRCA1 mRNA expression as a marker of time to progression and overall survival in sporadic breast cancers treated with anthracycline-based chemotherapy. These findings can be useful for customizing chemotherapy.     

Of Cattle,Sand Flies and Men: A Systematic Review of Risk Factor Analyses for South Asian Visceral Leishmaniasis and Implications for Elimination

Background

Studies performed over the past decade have identified fairly consistent epidemiological patterns of risk factors for visceral leishmaniasis (VL) in the Indian subcontinent.

Methods and Principal Findings

To inform the current regional VL elimination effort and identify key gaps in knowledge, we performed a systematic review of the literature, with a special emphasis on data regarding the role of cattle because primary risk factor studies have yielded apparently contradictory results. Because humans form the sole infection reservoir, clustering of kala-azar cases is a prominent epidemiological feature, both at the household level and on a larger scale. Subclinical infection also tends to show clustering around kala-azar cases. Within villages, areas become saturated over a period of several years; kala-azar incidence then decreases while neighboring areas see increases. More recently, post kala-azar dermal leishmaniasis (PKDL) cases have followed kala-azar peaks. Mud walls, palpable dampness in houses, and peri-domestic vegetation may increase infection risk through enhanced density and prolonged survival of the sand fly vector. Bed net use, sleeping on a cot and indoor residual spraying are generally associated with decreased risk. Poor micronutrient status increases the risk of progression to kala-azar. The presence of cattle is associated with increased risk in some studies and decreased risk in others, reflecting the complexity of the effect of bovines on sand fly abundance, aggregation, feeding behavior and leishmanial infection rates. Poverty is an overarching theme, interacting with individual risk factors on multiple levels.

Conclusions

Carefully designed demonstration projects, taking into account the complex web of interconnected risk factors, are needed to provide direct proof of principle for elimination and to identify the most effective maintenance activities to prevent a rapid resurgence when interventions are scaled back. More effective, short-course treatment regimens for PKDL are urgently needed to enable the elimination initiative to succeed.     

Histological and Physiological Characterization of Rind Breakdown of 'Navelate' Sweet Orange

Rind breakdown of ‘Navelate’ sweet orange is characterizedby sunken colourless areas of the peel which develop into reddish-brown,dry areas partially covering the exposed portion of the maturefruit. Sudden changes in relative humidity at fruit colour breakseem to be responsible for the natural development of this disorder,which begins at the transitional zone of the flavedo-albedoand advances across the flavedo reaching the epidermis. Affectedcells have reduced amounts of cytoplasm located in a centralposition and have twisted and squashed walls, forming areasof collapsed cells amongst the healthy cells of the flavedoand albedo. Comparisons of healthy and damaged areas of affectedfruits showed no significant differences in wax morphology andcuticular thickness or permeability. Our results suggest thatan excessive loss of water from hypodermal and albedo cellsis responsible for the disorder. Copyright 2001 Annals of BotanyCompany Citrus, physiological disorder, cell collapse, rind blemish, fruit quality     

Methodology of Clinical Trials Aimed at Assessing Interventions for Cutaneous Leishmaniasis

The current evidence-base for recommendations on the treatment of cutaneous leishmaniasis (CL) is generally weak. Systematic reviews have pointed to a general lack of standardization of methods for the conduct and analysis of clinical trials of CL, compounded with poor overall quality of several trials. For CL, there is a specific need for methodologies which can be applied generally, while allowing the flexibility needed to cover the diverse forms of the disease. This paper intends to provide clinical investigators with guidance for the design, conduct, analysis and report of clinical trials of treatments for CL, including the definition of measurable, reproducible and clinically-meaningful outcomes. Having unified criteria will help strengthen evidence, optimize investments, and enhance the capacity for high-quality trials. The limited resources available for CL have to be concentrated in clinical studies of excellence that meet international quality standards.     

Spatial and temporal evolution of distal 10q deletion,a prognostically unfavorable event in diffuse low-grade gliomas

Background

The disease course of patients with diffuse low-grade glioma is notoriously unpredictable. Temporal and spatially distinct samples may provide insight into the evolution of clinically relevant copy number aberrations (CNAs). The purpose of this study is to identify CNAs that are indicative of aggressive tumor behavior and can thereby complement the prognostically favorable 1p/19q co-deletion.

Results

Genome-wide, 50 base pair single-end sequencing was performed to detect CNAs in a clinically well-characterized cohort of 98 formalin-fixed paraffin-embedded low-grade gliomas. CNAs are correlated with overall survival as an endpoint. Seventy-five additional samples from spatially distinct regions and paired recurrent tumors of the discovery cohort were analyzed to interrogate the intratumoral heterogeneity and spatial evolution. Loss of 10q25.2-qter is a frequent subclonal event and significantly correlates with an unfavorable prognosis. A significant correlation is furthermore observed in a validation set of 126 and confirmation set of 184 patients. Loss of 10q25.2-qter arises in a longitudinal manner in paired recurrent tumor specimens, whereas the prognostically favorable 1p/19q co-deletion is the only CNA that is stable across spatial regions and recurrent tumors.

Conclusions

CNAs in low-grade gliomas display extensive intratumoral heterogeneity. Distal loss of 10q is a late onset event and a marker for reduced overall survival in low-grade glioma patients. Intratumoral heterogeneity and higher frequencies of distal 10q loss in recurrences suggest this event is involved in outgrowth to the recurrent tumor.

Electronic supplementary material

The online version of this article (doi:10.1186/s13059-014-0471-6) contains supplementary material, which is available to authorized users.     

How Far Are We from Visceral Leishmaniasis Elimination in Bangladesh? An Assessment of Epidemiological Surveillance Data

Introduction

In 2005, Bangladesh, India, and Nepal joined forces to eliminate Visceral Leishmaniasis (or kala-azar) from the region by 2015. In Bangladesh the elimination target is set at less than one new case per 10,000 population per year at upazila (sub-district) level. As the deadline approaches, we review the status of the elimination initiative in this country.

Methods

We collected all available disease surveillance data at the Disease Control Unit of the Directorate General of Health Services, Government of Bangladesh from 1994 to 2013. Additionally, we retrieved data from the Civil Surgeon Office from the Mymensingh district, one of the most heavily affected areas in Bangladesh.

Results

Between 1994 and 2013, 109,266 kala-azar cases causing 329 deaths were reported from 37 endemic districts in Bangladesh. Only 16 districts reported cases every year. The Mymensingh district was the most affected with 53,582 (49.04%) cases. Between 2008 and 2013 only 16 upazilas showed incidence rates above the elimination target in which they ranged from 1.06 to 18.25 per 10,000 people per year.

Discussion

While clear progress has been made towards eliminating VL, 16 upazilas in Bangladesh had not yet reached the target in 2013, based on official notification data that probably suffered from under-reporting bias. The elimination initiative urgently needs to establish methods to ascertain and monitor the elimination target.