Microarray-based ultra-high resolution discovery of genomic deletion mutations

BACKGROUND: Oligonucleotide microarray-based comparative genomic hybridization (CGH) offers an attractive possible route for the rapid and cost-effective genome-wide discovery of deletion mutations. CGH typically involves comparison of the hybridization intensities of genomic DNA samples with microarray chip representations of entire genomes, and has widespread potential application in experimental research and medical diagnostics. However, the power to detect small deletions is low. RESULTS: Here we use a graduated series of Arabidopsis thaliana genomic deletion mutations (of sizes ranging from 4 bp to ~5 kb) to optimize CGH-based genomic deletion detection. We show that the power to detect smaller deletions (4, 28 and 104 bp) depends upon oligonucleotide density (essentially the number of genome-representative oligonucleotides on the microarray chip), and determine the oligonucleotide spacings necessary to guarantee detection of deletions of specified size. CONCLUSIONS: Our findings will enhance a wide range of research and clinical applications, and in particular will aid in the discovery of genomic deletions in the absence of a priori knowledge of their existence

Milking Characteristics of Istrian and Littoral Dinaric Donkey Breeds

Istrian and Littoral Dinaric donkey are autochthonous breeds which habitat primarily in the Mediterranean part of Croatia. During the second half of the 20th century they have lost primary function as working animals, which led to breeds suppression. Nowadays these two breeds are in the status of endangered ones. One of the possibilities for their economic re-affirmation is launching the program for production and processing donkey’s milk. The aim of the study was to determine the possibility of milk production of Istrian and Littoral Dinaric jennies in terms of quantity and chemical composition. Istrian jennies produce 745.4 mL/milking while Littoral Dinaric produce 317.8 mL/milking. Milk form Littoral Dinaric jennies contained a significantly higher proportion of milk fat and proteins. There were no significant differences in the content of lactose, dry matter, number of somatic cells and microorganisms in milk between these two breeds. Considering the potential for milk production and milk quality, we believe that both breeds are suitable for integration in the programs for milk production, thus enabling for both population to be economically sustainable. It is necessary to develop production technology and markets, with special emphasis considering quality of donkey milk and safety of consumers

Outcomes of transnasal endoscopic repair of cerebrospinal fluid leaks: a prospective cohort study

Abstract Background Although cerebrospinal fluid (CSF) leak repair of the anterior and middle skull base defect by endonasal endoscopic surgery (EES) presents one of the more difficult challenges, it has shown high success rates with less morbidity. Our objective is to evaluate the outcomes of transnasal endoscopic repair of CSF leak regarding success rate, impact on olfaction, and sinonasal function. Methods A prospective cohort study was conducted to evaluate the CSF leak repair outcomes related to the site, size of the defect, surgical techniques, and the materials that been used through Smell Identification Test (SIT), 22-item Sino-Nasal Outcome Test (SNOT-22), Perioperative Sinus Endoscopy score (POSE), and Lund-MacKay Scoring (LM) of CT scan. Results Twenty-one patients were enrolled in the study; 12 out of 21 were females with a higher prevalence of traumatic causes of 61.9%. Different techniques and materials were used for the repair with a success rate recorded at 90.5% after the first closure attempt. The mean standard deviation (SD) scores postoperatively (after 6 months) was markedly decreased in SNOT 22 with mean (SD) 5.55 ± 3.6, slightly increase in POSE (mean ± SD = 0.43 ± 0.6), and slightly decrease in SIT (mean ± SD =10.31 ± 4.7) and LM (mean ± SD = 0.57 ± 0.7). Conclusion Transnasal endoscopic CSF leak repair is an effective technique for skull base defect closure with a high success rate and no valuable morbidity to sinonasal function other than mild hyposmia in patients where nasoseptal (NSF) and septal flap have been used. Trial registration The study was approved by the institutional review board and ethics committee of (The Arab Board of Health Specializations) with order no. (453) on 1April 2018

Effects of FSH on testicular mRNA transcript levels in the hypogonadal mouse

FSH acts through the Sertoli cell to ensure normal testicular development and function. To identify transcriptional mechanisms through which FSH acts in the testis, we have treated gonadotrophin-deficient hypogonadal (hpg) mice with recombinant FSH and measured changes in testicular transcript levels using microarrays and real-time PCR 12, 24 and 72 h after the start of treatment. Approximately 400 transcripts were significantly altered at each time point by FSH treatment. At 12 h, there was a clear increase in the levels of a number of known Sertoli cell transcripts (e.g. Fabp5, Lgals1, Tesc, Scara5, Aqp5). Additionally, levels of Leydig cell transcripts were also markedly increased (e.g. Ren1, Cyp17a1, Akr1b7, Star, Nr4a1). This was associated with a small but significant rise in testosterone at 24 and 72 h. At 24 h, androgen-dependent Sertoli cell transcripts were up-regulated (e.g. Rhox5, Drd4, Spinlw1, Tubb3 and Tsx) and this trend continued up to 72 h. By contrast with the somatic cells, only five germ cell transcripts (Dkkl1, Hdc, Pou5f1, Zfp541 and 1700021K02Rik) were altered by FSH within the time-course of the experiment. Analysis of canonical pathways showed that FSH induced a general decline in transcripts related to formation and regulation of tight junctions. Results show that FSH acts directly and indirectly to induce rapid changes in Sertoli cell and Leydig cell transcript levels in the hpg mouse but that effects on germ cell development must occur over a longer time-span

A PRELIMINARY INVESTIGATION TO THE POSSIBLE IMPACT OF EL NINO/SOUTHERN OSCILLATION (ENSO) PHENOMENA ON THE CLIMATE OF TRINIDAD AND TOBAGO USING REMOTELY SENSED DATA

Abstract Trinidad and Tobago like most other small developing island states is quite vulnerable to changes in Climate and Sea Surface Temperature (SST). Climate changes can affect the rainfall regimes, air temperature, soil moisture budgets, water resources and arable land usage. While changes in air and sea surface temperature can adversely affect coral reefs, mangroves, sea grass bed communities and fishery populations that depend on them for habitat and breeding grounds. Analysis of the temporal and spatial changes of SST over the years is an important way of understanding the present variability in the climate. This will then allow for an improved assessment of the susceptibility, vulnerability and risk that both humans and natural ecosystems may face because of the changes in the coupling interaction of the various components of the Earth's environmental system. However, decision-making, management and developing environmental policies in this regard is seriously compromised due information poverty, i.e. the lack of data of effective and reliable information base. This paper advances a methodology to endure information poverty and to obtain and analyse the SST patterns utilising it as a proxy for determining the climate variability of Trinidad and Tobago. The methodology is based on obtaining, using and analysing available remotely sensed SST from the National Oceanographic and Atmospheric Agency (NOAA) via the Internet. As well as using available rainfall data. In the case of Trinidad and Tobago, the SST and rainfall data was obtained for the last 16 years. Mean monthly values of SST and rainfall for the period 1986-1999 were then derived. The results indicate that variability in the climate is occurring. Furthermore, it shows that during the El Niño years 1986-87 and 1997-98, the SST were at a maximum while the rainfall fell to minimum. The preliminary results indicated the possibility of developing statistical associations between SST and land precipitation as well as the association between the SST-land precipitation pattern and external events such as the El Niño Southern Oscillation (ENSO) phenomenon. These ideas in turn can lead to investigating the association between the variability in SST and land precipitation on water resources and land use patterns that might lead to the possible prediction of the land precipitation patterns based on SST analysis

Development and pilot implementation of a novel protocol to assess capacity and readiness of health systems to adopt HPV detection-based cervical cancer screening in Europe

BACKGROUND: Cervical cancer remains a significant public health concern in Europe. Effective introduction and scaling up of human papillomavirus (HPV) detection-based cervical cancer screening (CCS) requires a systematic assessment of health systems capacity. However, there is no validated capacity assessment methodology for CCS programmes, especially in European contexts. Addressing this gap, our study introduces an innovative and adaptable protocol for evaluating the capacity of CCS programmes across varying European health system settings. METHODS: Our research team developed a three-step capacity assessment framework, incorporating a health policy review checklist, a facility visit survey, and key informants' interview guide followed by a strengths, weaknesses, opportunities and threats (SWOT) analysis. Piloting this comprehensive approach, we explored the CCS capacity in three countries: Estonia, Portugal and Romania. These countries were selected due to their contrasting healthcare structures and resources, providing a diverse overview of the European context. RESULTS: Conducted over a period of 9 months, the capacity assessment covered multiple resources, 27 screening centres, 16 colposcopy and treatment centres and 15 key informant interviews. Our analysis highlighted both shared and country-specific challenges. A key common issue was ensuring high compliance to follow-up and management of screen-positive women. We identified considerable heterogeneity in resources and organization across the three countries, underscoring the need for tailored, rather than one-size-fits-all, solutions. CONCLUSIONS: Our study's novelty lies in the successful development of this capacity assessment methodology implementable within a relatively short time frame, proving its feasibility for use in various contexts and countries. The resulting set of materials, adaptable to different cancer types, is a ready-to-use toolkit to improve cancer screening processes and outcomes. This research marks a significant stride towards comprehensive capacity assessment for CCS programmes in Europe. Future directions include deploying these tools in other countries and cancer types, thereby contributing to the global fight against cancer. © 2024. The Author(s).CBIG-SCREEN has received funding from the EU Horizon 2020 Research and Innovation Programme under grant agreement no. 964049. Official starting date: 1 March 2021

Apolipoprotein E4 has extensive conformational heterogeneity in lipid-free and lipid-bound forms

The ε4-allele variant of apolipoprotein E (ApoE4) is the strongest genetic risk factor for Alzheimer\u27s disease, although it only differs from its neutral counterpart ApoE3 by a single amino acid substitution. While ApoE4 influences the formation of plaques and neurofibrillary tangles, the structural determinants of pathogenicity remain undetermined due to limited structural information. Previous studies have led to conflicting models of the C-terminal region positioning with respect to the N-terminal domain across isoforms largely because the data are potentially confounded by the presence of heterogeneous oligomers. Here, we apply a combination of single-molecule spectroscopy and molecular dynamics simulations to construct an atomically detailed model of monomeric ApoE4 and probe the effect of lipid association. Importantly, our approach overcomes previous limitations by allowing us to work at picomolar concentrations where only the monomer is present. Our data reveal that ApoE4 is far more disordered and extended than previously thought and retains significant conformational heterogeneity after binding lipids. Comparing the proximity of the N- and C-terminal domains across the three major isoforms (ApoE4, ApoE3, and ApoE2) suggests that all maintain heterogeneous conformations in their monomeric form, with ApoE2 adopting a slightly more compact ensemble. Overall, these data provide a foundation for understanding how ApoE4 differs from nonpathogenic and protective variants of the protein

Synthesis of Fluorine-18 Functionalized Nanoparticles for use as in vivo Molecular Imaging Agents

Nanoparticles containing fluorine-18 were prepared from block copolymers made by ring opening metathesis polymerization (ROMP). Using the fast initiating ruthenium metathesis catalyst (H_2IMes)(pyr)_2(Cl)_2Ru=CHPh, low polydispersity amphiphilic block copolymers were prepared from a cinnamoyl-containing hydrophobic norbornene monomer and a mesyl-terminated PEG-containing hydrophilic norbornene monomer. Self-assembly into micelles and subsequent cross-linking of the micelle cores by light-activated dimerization of the cinnamoyl groups yielded stable nanoparticles. Incorporation of fluorine-18 was achieved by nucleophilic displacement of the mesylates by the radioactive fluoride ion with 31% incorporation of radioactivity. The resulting positron-emitting nanoparticles are to be used as in vivo molecular imaging agents for use in tumor imaging

Clinical Profile of Cardiac Involvement in Danon Disease: A Multicenter European Registry

Background: The X-linked Danon disease manifests by severe cardiomyopathy, myopathy, and neuropsychiatric problems. We designed this registry to generate a comprehensive picture of clinical presentations and outcome of patients with Danon disease in cardiomyopathy centers throughout Europe. Methods: Clinical and genetic data were collected in 16 cardiology centers from 8 European countries. Results: The cohort comprised 30 male and 27 female patients. The age at diagnosis was birth to 42 years in men and 2 to 65 in women. Cardiac involvement was observed in 96%. Extracardiac manifestations were prominent in men but not in women. Left ventricular (LV) hypertrophy was reported in 73% of male and 74% of female patients. LV systolic dysfunction was reported in 40% of men (who had LV ejection fraction, 34±11%) and 59% of women (LV ejection fraction, 28±13%). The risk of arrhythmia and heart failure was comparable among sexes. The age of first heart failure hospitalization was lower in men (18±6 versus 28±17 years; P<0.003). Heart failure was the leading cause of death (10 of 17; 59%), and LV systolic dysfunction predicted an adverse outcome. Eight men and 8 women (28%) underwent heart transplantation or received an LV assist device. Our cohort suggests better prognosis of female compared with male heart transplant recipients. Conclusions: Danon disease presents earlier in men than in women and runs a malignant course in both sexes, due to cardiac complications. Cardiomyopathy features, heart failure and arrhythmia, are similar among the sexes. Clinical diagnosis and management is extremely challenging in women due to phenotypic diversity and the absence of extracardiac manifestations

Prenatal attachment: using measurement invariance to test the validity of comparisons across eight culturally diverse countries

Studies in high-income countries (HICs) have shown that variability in maternal-fetal attachment (MFA) predict important maternal health and child outcomes. However, the validity of MFA ratings in low- and middle-income countries (LMICs) remains unknown. Addressing this gap, we assessed measurement invariance to test the conceptual equivalence of the Prenatal Attachment Inventory (PAI: Muller, 1993) across eight LMICs. Our aim was to determine whether the PAI yields similar information from pregnant women across different cultural contexts. We administered the 18-item PAI to 1181 mothers in the third trimester (Mean age = 28.27 years old, SD = 5.81 years, range = 18–48 years) expecting their first infant (n = 359) or a later-born infant (n = 820) as part of a prospective birth cohort study involving eight middle-income countries: Ghana, Jamaica, Pakistan, Philippines, Romania, South Africa, Sri Lanka and Vietnam. We used Multiple Group Confirmatory Factor Analyses to assess across-site measurement invariance. A single latent factor with partial measurement invariance was found across all sites except Pakistan. Group comparisons showed that mean levels of MFA were lowest for expectant mothers in Vietnam and highest for expectant mothers in Sri Lanka. MFA was higher in first-time mothers than in mothers expecting a later-born child. The PAI yields similar information about MFA across culturally distinct middle-income countries. These findings strengthen confidence in the use of the tool across different settings; future studies should explore the use of the PAI as a screen for maternal behaviour that place children at risk