From images to insights: a neuroradiologist's practical guide on white matter fiber tract anatomy and DTI patterns for pre-surgical planning.

INTRODUCTION: Diffusion tensor imaging (DTI) is a valuable non-invasive imaging modality for mapping white matter tracts and assessing microstructural integrity, and can be used as a "biomarker" in diagnosis, differentiation, and therapeutic monitoring. Although it has gained clinical importance as a marker of neuropathology, limitations in its interpretation underscore the need for caution. METHODS: This review provides an overview of the principles and clinical applicability of DTI. We focus on major white matter fiber bundles, detailing their normal anatomy and pathological DTI patterns, with emphasis on tracts routinely requested in our neurosurgical department in the preoperative context (uncinate fasciculus, arcuate fasciculus, pyramidal pathway, optic radiation, and dentatorubrothalamic tract). RESULTS: We guide neuroradiologists and neurosurgeons in defining volumes of interest for mapping individual tracts and demonstrating their 3D reconstructions. The intricate trajectories of white matter tracts pose a challenge for accurate fiber orientation recording, with each bundle exhibiting specific characteristics. Tracts adjacent to brain lesions are categorized as displaced, edematous, infiltrated, or disrupted, illustrated with clinical cases of brain neoplasms. To improve structured reporting, we propose a checklist of topics for inclusion in imaging evaluations and MRI reports. CONCLUSION: DTI is emerging as a powerful tool for assessing microstructural changes in brain disorders, despite some challenges in standardization and interpretation. This review serves an educational purpose by providing guidance for fiber monitoring and interpretation of pathological patterns observed in clinical cases, highlighting the importance and potential pitfalls of DTI in neuroradiology and surgical planning.

The Role of Natural and Synthetic Flavonoids in the Prevention of Marine Biofouling.

Marine biofouling is a major concern for the maritime industry, environment, and human health. Biocides which are currently used in marine coatings to prevent this phenomenon are toxic to the marine environment, and therefore a search for antifoulants with environmentally safe properties is needed. A large number of scientific papers have been published showing natural and synthetic compounds with potential to prevent the attachment of macro- and microfouling marine organisms on submerged surfaces. Flavonoids are a class of compounds which are highly present in nature, including in marine organisms, and have been found in a wide range of biological activities. Some natural and synthetic flavonoids have been evaluated over the last few years for their potential to prevent the settlement and/or the growth of marine organisms on submerged structures, thereby preventing marine biofouling. This review compiles, for the first-time, natural flavonoids as well as their synthetic analogues with attributed antifouling activity against macrofouling and microfouling marine organisms.

Proof of Concept of Natural and Synthetic Antifouling Agents in Coatings.

Marine biofouling, caused by the deposition and accumulation of marine organisms on submerged surfaces, represents a huge concern for the maritime industries and also contributes to environmental pollution and health concerns. The most effective way to prevent this phenomenon is the use of biocide-based coatings which have proven to cause serious damage to marine ecosystems. Several research groups have focused on the search for new environmentally friendly antifoulants, including marine and terrestrial natural products and synthetic analogues. Some of these compounds have been incorporated into marine coatings and display interesting antifouling activities caused by the interference with the biofilm-forming species as well as by the inhibition of the settlement of macroorganisms. This review highlights the proof-of-concept studies of emerging natural or synthetic antifouling compounds in coatings, from lab-made to commercial ones, performed between 2019 and 2023 and their results in the field or in in vivo laboratorial tests.

Leishmania (Leishmania) infantum DNA detection in Nyssomyia neivai in Vale do Ribeira, Paraná, Brazil.

BACKGROUND: The incidence of visceral leishmaniasis (VL) has increased in the Southern region of Brazil in recent years, especially in the State of Paraná. New species have been suggested with potential to act as vector in VL endemic areas. OBJECTIVES: Identify the Leishmania species in sand fly specimens collected from 2016 to 2018 in the municipality of Itaperuçu, Vale do Ribeira, Paraná, Brazil. METHODS: Light traps were used for collections and for the analysis of sand fly were used the multiplex polymerase chain reaction (PCR) methodology and subsequent sequencing. FINDINGS: Among the collected specimens, 88.62% were attributed to the species Nyssomyia neivai, which were grouped into 176 pools. Three positive pools were detected: two with Leishmania (Viannia) braziliensis and one with L. (Leishmania) infantum. The positivity rate for the parasite was 0.25% based on the presence of at least one infected insect in the pool. MAIN CONCLUSIONS: The detection of L. infantum in Ny. neivai draws attention due to its abundance and anthropophily in the State of Paraná. Moreover, this finding is considered as an alert and suggests that the vector competence of Ny. neivai and the criteria for its incrimination should be carried out, given its wide distribution in southern of Brazil.

Characterisation of an area of coexistent visceral and cutaneous leishmaniasis transmission in the State of Piauí, Brazil.

BACKGROUND: In Brazil, transmission of visceral and cutaneous leishmaniasis has expanded geographically over the last decades, with both clinical forms occurring simultaneously in the same area. OBJECTIVES: This study characterised the clinical, spatial, and temporal distribution, and performed entomological surveillance and natural infection analysis of a leishmaniasis-endemic area. METHODS: In order to characterise the risk of leishmaniasis transmission in Altos, Piauí, we described the clinical and socio-demographic variables and the spatial and temporal distribution of cases of American visceral leishmaniasis (AVL) and American cutaneous leishmaniasis (ACL) cases and identified potential phlebotomine vectors. FINDINGS: The urban area concentrated almost 54% of ACL and 86.8% of AVL cases. The temporal and spatial distribution of AVL and ACL cases in Altos show a reduction in the number of risk areas, but the presence of permanent disease transmission foci is observed especially in the urban area. 3,808 phlebotomine specimens were captured, with Lutzomyia longipalpis as the most frequent species (98.45%). Of the 35 females assessed for natural infection, one specimen of Lu. longipalpis tested positive for the presence of Leishmania infantum and Leishmania braziliensis DNA. MAIN CONCLUSION: Our results indicate the presence of risk areas for ACL and AVL in the municipality of Altos and highlight the importance of entomological surveillance to further understand a possible role of Lu. longipalpis in ACL transmission.

Chalcone derivatives as promising antifoulants: Molecular optimization, bioactivity evaluation and performance in coatings.

Marine biofouling remains a huge concern for maritime industries and for environmental health. Although the current biocide-based antifouling coatings can prevent marine biofouling, their use has been associated with toxicity for the marine environment, being urgent to find sustainable alternatives. Previously, our research group has identified a prenylated chalcone (1) with promising antifouling activity against the settlement of larvae of the macrofouling species Mytilus galloprovincialis (EC50 = 16.48 µM and LC50 > 200 µM) and lower ecotoxicity when compared to Econea®, a commercial antifouling agent in use. Herein, a series of chalcone 1 analogues were designed and synthesized in order to obtain optimized antifouling compounds with improved potency while maintaining low ecotoxicity. Compounds 8, 15, 24, and 27 showed promising antifouling activity against the settlement of M. galloprovincialis larvae, being dihydrochalcone 27 the most potent. The effect of compound 24 was associated with the inhibition of acetylcholinesterase activity. Among the synthesized compounds, compound 24 also showed potent complementary activity against Navicula sp. (EC50 = 4.86 µM), similarly to the lead chalcone 1 (EC50 = 6.75 µM). Regarding the structure-activity relationship, the overall results demonstrate that the substitution of the chalcone of the lead compound 1 by a dihydrochalcone scaffold resulted in an optimized potency against the settlement of mussel larvae. Marine polyurethane (PU)-based coatings containing the best performed compound concerning anti-settlement activity (dihydrochalcone 27) were prepared, and mussel larvae adherence was reduced compared to control PU coatings.

ChatGPT as an item calibration tool: Psychometric insights in a high-stakes examination.

INTRODUCTION: ChatGPT has attracted a lot of interest worldwide for its versatility in a range of natural language tasks, including in the education and evaluation industry. It can automate time- and labor-intensive tasks with clear economic and efficiency gains. METHODS: This study evaluated the potential of ChatGPT to automate psychometric analysis of test questions from the 2020 Portuguese National Residency Selection Exam (PNA). ChatGPT was queried 100 times with the 150 MCQ from the exam. Using ChatGPT's responses, difficulty indices were calculated for each question based on the proportion of correct answers. The predicted difficulty levels were compared to the actual difficulty levels of the 2020 exam MCQ's using methods from classical test theory. RESULTS: ChatGPT's predicted item difficulty indices positively correlated with the actual item difficulties (r (148) = -0.372, p < .001), suggesting a general consistency between the real and the predicted values. There was also a moderate significant negative correlation between the difficulty index predicted by ChatGPT and the number of challenges (r (148) = -0.302, p < .001), highlighting ChatGPT's potential for identifying less problematic questions. CONCLUSION: These findings unveiled ChatGPT's potential as a tool for assessment development, proving its capability to predict the psychometric characteristics of high-stakes test items in automated item calibration without pre-testing in real-life scenarios.

Effect of a Non-pharmacological Intervention on Vaccine-related Pain: Randomized Clinical Trial.

BACKGROUND: Acute pain during vaccine administration is an expected event; however, some strategies, such as the use of high frequency vibration devices associated with cryotherapy, may minimize it. AIM: Evaluate the effect of high frequency vibration associated with cryotherapy on the level of pain related to the administration of influenza vaccine by intramuscular route in adults. METHOD: A randomized clinical trial was conducted with 350 adults who received the influenza vaccine. Participants allocated to the intervention group used a high-frequency vibration device associated with cryotherapy during vaccination, and those in the control group administered the vaccine according to the service routine. RESULTS: Mean self-reported pain after vaccination of participants who used the device was lower (1.5 ±1.7) when compared to those who did not use it (1.9 ±1.9) (p = .041). There was an association of higher levels of pain after vaccination with participants in the control group (p = .011), females (p = .042), with higher level of pain expected with vaccination (p < .001) and higher level of anxiety before (p < .001) and after vaccination (p = .001). CONCLUSIONS: The use of high frequency vibration associated with cryotherapy has been shown to be a viable non-pharmacological intervention for the reduction of pain associated with influenza vaccination in adults.

Posterior semicircular canal cupulolithiasis during acute pontomedullary demyelination.

Positional vertigo poses a diagnostic challenge in people with multiple sclerosis (MS). The characteristics of positional nystagmus and its response to repositioning manoeuvres are usually sufficient to diagnose benign paroxysmal positional vertigo (BPPV). However, certain BPPV variants respond poorly to repositioning manoeuvres and their nystagmus pattern can resemble that of central positional vertigo caused by infratentorial demyelination. This diagnostic difficulty is particularly challenging if positional vertigo occurs during an MS relapse. We describe a woman with MS who developed a sixth nerve palsy and gaze-evoked nystagmus, caused by demyelination near or within areas classically involved in central positional vertigo. However, she also had positional vertigo from coincident BPPV (and not central positional vertigo). This was initially a treatment resistant-posterior semicircular canal cupulolithiasis but it later progressed to a posterior semicircular canal canalolithiasis, with symptoms promptly resolving after a repositioning manoeuvre.

Efferocytosis is restricted by axon guidance molecule EphA4 via ERK/Stat6/MERTK signaling following brain injury.

BACKGROUND: Efferocytosis is a process that removes apoptotic cells and cellular debris. Clearance of these cells alleviates neuroinflammation, prevents the release of inflammatory molecules, and promotes the production of anti-inflammatory cytokines to help maintain tissue homeostasis. The underlying mechanisms by which this occurs in the brain after injury remain ill-defined. METHODS: We used GFP bone marrow chimeric knockout (KO) mice to demonstrate that the axon guidance molecule EphA4 receptor tyrosine kinase is involved in suppressing MERTK in the brain to restrict efferocytosis of resident microglia and peripheral-derived monocyte/macrophages. RESULTS: Single-cell RNAseq identified MERTK expression, the primary receptor involved in efferocytosis, on monocytes, microglia, and a subset of astrocytes in the damaged cortex following brain injury. Loss of EphA4 on infiltrating GFP-expressing immune cells improved functional outcome concomitant with enhanced efferocytosis and overall protein expression of p-MERTK, p-ERK, and p-Stat6. The percentage of GFP+ monocyte/macrophages and resident microglia engulfing NeuN+ or TUNEL+ cells was significantly higher in KO chimeric mice. Importantly, mRNA expression of Mertk and its cognate ligand Gas6 was significantly elevated in these mice compared to the wild-type. Analysis of cell-specific expression showed that p-ERK and p-Stat6 co-localized with MERTK-expressing GFP + cells in the peri-lesional area of the cortex following brain injury. Using an in vitro efferocytosis assay, co-culturing pHrodo-labeled apoptotic Jurkat cells and bone marrow (BM)-derived macrophages, we demonstrate that efferocytosis efficiency and mRNA expression of Mertk and Gas6 was enhanced in the absence of EphA4. Selective inhibitors of ERK and Stat6 attenuated this effect, confirming that EphA4 suppresses monocyte/macrophage efferocytosis via inhibition of the ERK/Stat6 pathway. CONCLUSIONS: Our findings implicate the ERK/Stat6/MERTK axis as a novel regulator of apoptotic debris clearance in brain injury that is restricted by peripheral myeloid-derived EphA4 to prevent the resolution of inflammation.

Prototheca bovis in goats: Experimental mastitis and treatment with polypyrrole.

Prototheca bovis has been associated with several cases of mastitis in cattle but no record of intramammary infections has been reported in goats. This infection does not respond to available treatments and the disposal recommendation of affected animals cause great damage to the dairy industry. Alternatives for dealing with infections caused by Prototheca spp. are required worldwide. In vitro results suggest polypyrrole as promising molecule for combating this alga, because an algaecide effect was observed on tested Prototheca spp. isolates. Thus, this study evaluated goats as an experimental model for intramammary infection by P. bovis and a protocol for treating these animals with an intramammary polypyrrole solution. The possibility of P. bovis promoting an intramammary infection in goats was experimentally proven, demonstrating this species as an important model for studies involving algae mastitis. Furthermore, polypyrrole reduced the counts of Prototheca sp. in the analyzed samples, showing potential to fight this microorganism also in vivo. The results obtained in this study demonstrate the ability of P. bovis to colonize breast tissue in lactating goats and the highly soluble molecule of polypyrrole has potential use for the treatment of protothecosis.

Unusual Microsatellite-Instable Mixed Neuroendocrine and Non-neuroendocrine Neoplasm: A Clinicopathological Inspection and Literature Review.

BACKGROUND: Mixed neuroendocrine and non-endocrine neoplasms (MiNENs) are challenging to diagnose and manage clinically. The current understanding of MiNENs' pathobiology, molecular mechanisms, and management is incomplete. Though microsatellite instability (MSI) is known to impact carcinogenesis, reports examining MSI mechanisms for MiNENs are rare. METHODS: We report an unusual colonic MSI-MiNEN uncovered in an 89-year-old woman and the review of the literature. RESULTS: Pathologic inspection revealed a high-grade carcinoma composed of tumor cells with neuroendocrine histologic traits and immunophenotype intermixed with mucin-containing signet ring-like cells arranged in nested and micronodular patterns. Loss of MLH1 and PMS2 mismatch repair proteins was detected in tumor cells. INSM1 immunostaining highlighted about 50% of the tumour, further reinforcing the MiNEN diagnosis. Next-generation sequencing identified multiple carcinogenic mutations. Because of the advanced stage of the tumor and its adhesion to the adjacent organs, surgical resection was aborted; immunotherapy was initiated. The tumor is in remission 30 months following initiation of treatment, and the patient remains asymptomatic. CONCLUSION: This unique MSI MiNEN was characterized by its immunohistochemical and molecular signatures and illustrated how correctly diagnosing MSI can strongly improve a patient's outcomes.

Cryptococcosis in domestic and wild animals: A review.

Cryptococcosis is a fungal disease of public health relevance that affects numerous animal species and humans, causing respiratory and neurological impairment. Hence, we conducted a systematic review that included publications from 1975 to 2021 and covered 132 articles that addressed reports of cryptococcosis in domestic and wild animals, its main clinical manifestations, pathological findings, etiology, diagnosis, and therapeutic protocols. We found that the highest number of reports of cryptococcosis is in domestic species, especially cats. Among the wild and/or exotic animals, koalas and ferrets are the most affected, being important carriers of Cryptococcus spp. Pulmonary and neurological involvement is predominant in all species, although nonspecific clinical manifestations have been reported in various species, making clinical suspicion and diagnosis difficult. The countries with the most reports are Australia, the United States, Brazil, and Canada, with C. gattii VGI and VGII standing out. The therapies were based on azoles, amphotericin B, and 5-flucytosine, although there is no standard treatment protocol. Although, several diagnostic methods have been described, in a significant number of reports the diagnosis was made after a necropsy. Professionals are warned about diverse and nonspecific clinical manifestations in different animal species, which underlines the importance of cryptococcosis in the differential diagnosis in clinical practice. Furthermore, it is necessary to encourage the use of laboratory and molecular tools to improve the diagnosis of cryptococcosis. We also emphasize the urgent need for standardized therapeutic protocols to guide veterinary clinicians.

This review compiles studies on cryptococcosis in domestic and wild animals. Most reports occurred in cats and koalas. Pulmonary and neurological involvement was predominant in all affected species, and C. gattii VGI and VGII stood out in the etiology of the disease.

The role of early functional neuroimaging in predicting neurodevelopmental outcomes in neonatal encephalopathy.

Reliably assessing the early neurodevelopmental outcomes in infants with neonatal encephalopathy (NE) is of utmost importance to advise parents and implement early and personalized interventions. We aimed to evaluate the accuracy of neuroimaging modalities, including functional magnetic resonance imaging (fMRI) in predicting neurodevelopmental outcomes in NE. Eighteen newborns with NE due to presumed perinatal asphyxia (PA) were included in the study, 16 of whom underwent therapeutic hypothermia. Structural magnetic resonance imaging (MRI), and fMRI during passive visual, auditory, and sensorimotor stimulation were acquired between the 10th and 14th day of age. Clinical follow-up protocol included visual and auditory evoked potentials and a detailed neurodevelopmental evaluation at 12 and 18 months of age. Infants were divided according to sensory and neurodevelopmental outcome: severe, moderate disability, or normal. Structural MRI findings were the best predictor of severe disability with an AUC close to 1.0. There were no good predictors to discriminate between moderate disability versus normal outcome. Nevertheless, structural MRI measures showed a significant correlation with the scores of neurodevelopmental assessments. During sensorimotor stimulation, the fMRI signal in the right hemisphere had an AUC of 0.9 to predict absence of cerebral palsy (CP). fMRI measures during auditory and visual stimulation did not predict sensorineural hearing loss or cerebral visual impairment. CONCLUSION: In addition to structural MRI, fMRI with sensorimotor stimulation may open the gate to improve the knowledge of neurodevelopmental/motor prognosis if proven in a larger cohort of newborns with NE. WHAT IS KNOWN: • Establishing an early, accurate neurodevelopmental prognosis in neonatal encephalopathy remains challenging. • Although structural MRI has a central role in neonatal encephalopathy, advanced MRI modalities are gradually being explored to optimize neurodevelopmental outcome knowledge. WHAT IS NEW: • Newborns who later developed cerebral palsy had a trend towards lower fMRI measures in the right sensorimotor area during sensorimotor stimulation. • These preliminary fMRI results may improve future early delineation of motor prognosis in neonatal encephalopathy.

A suggestive approach for assessing item quality, usability and validity of Automatic Item Generation.

Automatic Item Generation (AIG) refers to the process of using cognitive models to generate test items using computer modules. It is a new but rapidly evolving research area where cognitive and psychometric theory are combined into digital framework. However, assessment of the item quality, usability and validity of AIG relative to traditional item development methods lacks clarification. This paper takes a top-down strong theory approach to evaluate AIG in medical education. Two studies were conducted: Study I-participants with different levels of clinical knowledge and item writing experience developed medical test items both manually and through AIG. Both item types were compared in terms of quality and usability (efficiency and learnability); Study II-Automatically generated items were included in a summative exam in the content area of surgery. A psychometric analysis based on Item Response Theory inspected the validity and quality of the AIG-items. Items generated by AIG presented quality, evidences of validity and were adequate for testing student's knowledge. The time spent developing the contents for item generation (cognitive models) and the number of items generated did not vary considering the participants' item writing experience or clinical knowledge. AIG produces numerous high-quality items in a fast, economical and easy to learn process, even for inexperienced and without clinical training item writers. Medical schools may benefit from a substantial improvement in cost-efficiency in developing test items by using AIG. Item writing flaws can be significantly reduced thanks to the application of AIG's models, thus generating test items capable of accurately gauging students' knowledge.

Protothecosis in Veterinary Medicine: a minireview.

Protothecosis is an important infection caused by Prototheca spp., an alga that affect humans and animals. Prototheca spp. infections determine losses of production and quality of life for the affected animals. In this disease, prevention and early diagnosis are essential to avoid the spread of the agent to susceptible hosts. This review aimed to assemble the reported cases of protothecosis in Veterinary Medicine, highlighting Prototheca species involved in the infections, the main animal species affected, as well as the clinical manifestations, diagnosis, and treatment. Our findings showed that protothecosis has been reported in various domestic and wild animal species, presenting diverse clinical manifestations, including mastitis in cows, respiratory manifestations in goats and cats, and a wide range of clinical signs in dogs. The clinical diagnosis and treatment of Prototheca spp. infections are difficult and infected animals are frequently discarded or euthanized. Due to the importance of this infection, protothecosis must be considered an important differential diagnosis in routine clinical Veterinary Medicine.

Oomicidal activity of polypyrrole nanoparticles against Pythium insidiosum.

This study evaluated in-vitro action of a new molecule, the polypyrrole nanoparticles (Ppy-NP), against Pythium insidiosum isolates using M38-A2/CLSI; the minimal inhibitory (MIC) and minimal oomicidal (MOC) concentrations were also determined. Additionally, changes in the hyphae wall of P. insidiosum CBS 575.85 treated with Ppy-NP were evaluated by scanning electron microscopy (SEM). The MIC100 and MOC for all isolates ranged from 8 to 32 µg mL-1, and the MIC90 and MIC50 were 16 µg mL-1. The SEM showed structural damage to the hyphae of P. insidisoum treated with Ppy-NP, as hyphae surfaces with less turgidity were found, thereby showing scaling and ruptures compared to the control (untreated hyphae). Our findings highlighted the anti-P. insidiosum properties of Ppy-NP proved to be a promising candidate for research using pythiosis experimental models.

Anti-Pythium insidiosum activity of bioactive compounds from medicinal plants.

Pythiosis is a serious disease caused by the aquatic oomycete Pythium insidiosum that mainly affects mammals. Unlike fungal and bacterial resistance induced by the indiscriminate use of drugs, P. insidiosum has low susceptibility to antifungal drugs. In this sense, essential oils and their major components emerge as a promising treatment line for this disease. Given the above, this study sought to verify P. insidiosum (n = 34) susceptibility to the bioactive compounds eugenol, α-terpineol, menthol, and carvacrol and correlate them with the respective essential oils of Eugenia caryophyllata, Melaleuca alternifolia, Mentha piperita, and Origanum vulgare. The essential oils and bioactive compounds were purchased commercially and tested according to the Clinical and Laboratory Standards Institute protocol M38-A2. Our findings showed that eugenol, α-terpineol, and carvacrol had superior anti-P. insidiosum action than their respective essential oils, suggesting that they may be responsible for inhibitory activity against P. insidiosum. Notably, the major compound with the best anti-P. insidiosum activity was α-terpineol; nonetheless, menthol showed less activity than its essential oil. The results imply that essential oils and their major compounds may be important allies in treating pythiosis, expanding the perspectives of developing new drugs with anti-P. insidiosum activity.

Effect of use and dosage of p-follicle-stimulating hormone for ovarian superstimulation before ovum pick-up and in vitro embryo production in pregnant Holstein heifers.

The benefit of ovarian superstimulation using exogenous FSH before ovum pick-up (OPU) and in vitro embryo production (IVEP) has been the subject of conflicting results. The objective of the present study, therefore, was to evaluate the effect of use and dose of porcine FSH (p-FSH) before OPU/IVEP on ovarian response and embryo production in pregnant heifers. Pregnant Holstein heifers (n = 48) were randomly assigned to receive 0, 160, or 300 mg NIH-FSH-P1 in a crossover design. Ovum pick-up was performed at 49, 63, and 77 d of gestation with a 14 d "washout" between OPU sessions. Follicle ablation was performed on D 0 (p.m.) and p-FSH treatments, consisting of 4 decreasing dose injections administered 12 h apart, were initiated 36 h after follicle ablation (d 2 a.m.). Heifers underwent OPU on D 5 (a.m.), 40 h after the last p-FSH treatment, and cumulus oocyte complexes (COC) were subjected to IVEP procedures. Differences between treatment groups were evaluated using generalized linear mixed models. There were quadratic effects of treatment on both number and percentage of small (<6 mm), medium (6-10 mm), and large (>10 mm) follicles. Number and percentage of medium follicles increased with increasing p-FSH dosages, although the magnitude of the change was greater between 0 and 160 mg, than between 160 and 300 mg of p-FSH. Total number of follicles, number of COC recovered and number of viable COC increased linearly with increasing p-FSH dose. Conversely, there was no evidence for an effect of p-FSH dose on COC recovery percentage nor the percentage of viable COC. Cleavage percentage, number of cleaved oocytes, blastocyst percentage, and number of blastocysts increased linearly with increasing p-FSH dose. In conclusion, use of p-FSH before OPU resulted in greater superstimulatory response and oocyte competence which in turn increased IVEP. Furthermore, these effects were dose dependent such that use of a greater dose of p-FSH up to 300 mg progressively increased embryo yield.

HRM Accuracy and Limitations as a Species Typing Tool for Leishmania Parasites.

High Resolution Melting Analysis (HRM) has been pointed out as a suitable alternative method to detect and identify Leishmania species. Herein, we aimed to evaluate the sensitivity, specificity, accuracy, and limitations of a HSP70-HRM protocol both as a diagnostic scheme applied in clinical samples and as a species typing tool for laboratory research and reference services. Our data reveal the pronounced species-typing potential of the HSP70-HRM in DNA from cultured parasites. For clinical samples, however, we advise caution due to parasite load-dependent accuracy. In light of these findings and considering the importance of parasite load determination for clinical and research purposes, we recommend the integration of the presented typing scheme and the previously published Leishmania quantifying approach as combined tools for clinicians, surveillance, and research.