Sleep deprivation induces late deleterious effects in a pharmacological model of Parkinsonism.

Parkinson's disease is a degenerative, chronic and progressive disease, characterized by motor dysfunctions. Patients also exhibit non-motor symptoms, such as affective and sleep disorders. Sleep disorders can potentiate clinical and neuropathological features and lead to worse prognosis. The goal of this study was to evaluate the effects of sleep deprivation (SD) in mice submitted to a progressive pharmacological model of Parkinsonism (chronic administration with a low dose of reserpine). Male Swiss mice received 20 injections of reserpine (0.1 mg/kg) or vehicle, on alternate days. SD was applied before or during reserpine treatment and was performed by gentle handling for 6 h per day for 10 consecutive days. Animals were submitted to motor and non-motor behavioral assessments and neurochemical evaluations. Locomotion was increased by SD and decreased by reserpine treatment. SD during treatment delayed the onset of catalepsy, but SD prior to treatment potentiated reserpine-induced catalepsy. Thus, although SD induced an apparent beneficial effect on motor parameters, a delayed deleterious effect on alterations induced by reserpine was found. In the object recognition test, both SD and reserpine treatment produced cognitive deficits. In addition, the association between SD and reserpine induced anhedonic-like behavior. Finally, an increase in oxidative stress was found in hippocampus of mice subjected to SD, and tyrosine hydroxylase immunoreactivity was reduced in substantia nigra of reserpine-treated animals. Results point to a possible late effect of SD, aggravating the deficits in mice submitted to the reserpine progressive model of PD.

Transmission Route Used By Parasitic Lasidium Larvae of The Freshwater Mussel Anodontites Trapesialis on Guppies Poecilia Reticulata During Short Cohabitation.

We describe for the first time the transmission route employed by the parasitic larvae of the freshwater mussel Anodontites trapesialis (Lamarck, 1819) during cohabitation on the guppy Poecilia reticulata Peter, 1859. The freshwater mussel and fish-host were employed as a model system to investigate the infection and parasite establishment. Laboratory experiments included video recording and histopathological analysis of the infection. In vivo video observations demonstrated that lasidium larvae could be transmitted to guppies during direct contact. A series of histology samples and photography suggest that this larva attaches and colonizes possibly on-site on the outer surface of the fish, causing cellular inflammation in the epidermis layer with cellular hyperplasia in the zone of parasite attachment. An evident hyaline layer, cellular hypertrophy, and a large number of undifferentiated proliferating cells were observed. Hemorrhagic tissue and swelling were observed in the epidermis and dermic zone. The total number of larvae per male and female guppy was 525 ± 86 and 494 ± 167, respectively. No parasitic preference was detected for male versus female parasitized fish.

Prevalence of Parkinson's disease: a population-based study in Portugal.

BACKGROUND AND PURPOSE: Portugal has been identified as one of the countries with a high prevalence of LRRK2-G2019S, considered to be the most frequent known cause of familial and sporadic Parkinson's disease (PD). The aim of this study was to evaluate the prevalence of PD in Portugal using a door-to-door methodology. METHODS: A cross-sectional study was conducted in the Portuguese community-dwelling population; that is, elderly people living in the community on their own, aged ≥50 years and resident in mainland Portugal, in two phases: (i) a questionnaire was applied to screen potential cases of PD; and (ii) screened cases were evaluated by an expert in PD to confirm diagnosis. RESULTS: The adjusted prevalence of PD for the Portuguese community-dwelling population aged ≥50 years was 0.24%. The estimated total number of cases of PD for the Portuguese population is 180/100 000 inhabitants. CONCLUSIONS: The results of this study show that a geographical region with a high frequency of a causal mutation for PD does not automatically imply a high prevalence of patients with PD.

Warm vegetarians? Heat waves and diet shifts in tadpoles.

Temperature can play an important role in determining the feeding preferences of ectotherms. In light of the warmer temperatures arising with the current climatic changes, omnivorous ectotherms may perform diet shifts toward higher herbivory to optimize energetic intake. Such diet shifts may also occur during heat waves, which are projected to become more frequent, intense, and longer lasting in the future. Here, we investigated how heat waves of different duration affect feeding preferences in omnivorous anuran tadpoles and how these choices affect larval life history. In laboratory experiments, we fed tadpoles of three species on animal, plant, or mixed diet and exposed them to short heat waves (similar to the heat waves these species experience currently) or long heat waves (predicted to increase under climate change). We estimated the dietary choices of tadpoles fed on the mixed diet using stable isotopes and recorded tadpole survival and growth, larval period, and mass at metamorphosis. Tadpole feeding preferences were associated with their thermal background, with herbivory increasing with breeding temperature in nature. Patterns in survival, growth, and development generally support decreased efficiency of carnivorous diets and increased efficiency or higher relative quality of herbivorous diets at higher temperatures. All three species increased herbivory in at least one of the heat wave treatments, but the responses varied among species. Diet shifts toward higher herbivory were maladaptive in one species, but beneficial in the other two. Higher herbivory in omnivorous ectotherms under warmer temperatures may impact species differently and further contribute to changes in the structure and function of freshwater environments.

Optogenetic activation of intracellular adenosine A2A receptor signaling in the hippocampus is sufficient to trigger CREB phosphorylation and impair memory.

Human and animal studies have converged to suggest that caffeine consumption prevents memory deficits in aging and Alzheimer's disease through the antagonism of adenosine A2A receptors (A2ARs). To test if A2AR activation in the hippocampus is actually sufficient to impair memory function and to begin elucidating the intracellular pathways operated by A2AR, we have developed a chimeric rhodopsin-A2AR protein (optoA2AR), which retains the extracellular and transmembrane domains of rhodopsin (conferring light responsiveness and eliminating adenosine-binding pockets) fused to the intracellular loop of A2AR to confer specific A2AR signaling. The specificity of the optoA2AR signaling was confirmed by light-induced selective enhancement of cAMP and phospho-mitogen-activated protein kinase (p-MAPK) (but not cGMP) levels in human embryonic kidney 293 (HEK293) cells, which was abolished by a point mutation at the C terminal of A2AR. Supporting its physiological relevance, optoA2AR activation and the A2AR agonist CGS21680 produced similar activation of cAMP and p-MAPK signaling in HEK293 cells, of p-MAPK in the nucleus accumbens and of c-Fos/phosphorylated-CREB (p-CREB) in the hippocampus, and similarly enhanced long-term potentiation in the hippocampus. Remarkably, optoA2AR activation triggered a preferential p-CREB signaling in the hippocampus and impaired spatial memory performance, while optoA2AR activation in the nucleus accumbens triggered MAPK signaling and modulated locomotor activity. This shows that the recruitment of intracellular A2AR signaling in the hippocampus is sufficient to trigger memory dysfunction. Furthermore, the demonstration that the biased A2AR signaling and functions depend on intracellular A2AR loops prompts the possibility of targeting the intracellular A2AR-interacting partners to selectively control different neuropsychiatric behaviors.

Magnetic resonance correlation of iron content with neuromelanin in the substantia nigra of early-stage Parkinson's disease.

BACKGROUND AND PURPOSE: Magnetic resonance (MR) studies have demonstrated a significant reduction of neuromelanin in the substantia nigra (SN) of Parkinson's disease (PD) patients with high accuracy for differential diagnosis compared to non-PD controls and essential tremor. However, studies state that not knowing how paramagnetic effects of iron influence neuromelanin signal is a limitation. In this study a neuromelanin-sensitive MR sequence was combined with T2* relaxometry iron quantification analysis to study the SN of early-stage PD patients to investigate the correlation between these parameters. METHODS: The inclusion criteria were untreated de novo PD patients and a 2-5 year disease duration (early PD); in addition, age-matched controls were enrolled. These were studied at 3.0 T with a high-resolution T1-weighted MR sequence to visualize neuromelanin and a relaxometry sequence for iron quantification. The primary outcome was the correlation of the width of the neuromelanin high signal region and the T2* values in the lateral, central and medial segments of the SN. RESULTS: Very weak correlations of T2* values with neuromelanin width, positive for global and negative for the medial and lateral SN segments, were found in both PD groups and control subjects. The SN neuromelanin width was markedly reduced in the de novo and early PD groups compared with controls in all SN segments, but no significant difference in T2* values was found between the groups. CONCLUSIONS: The SN neuromelanin signal does not have a significant correlation with iron content in PD patients or controls. The neuromelanin MR signal reduction in PD does not seem to be significantly influenced by paramagnetic iron effects.

Substantia nigra neuromelanin magnetic resonance imaging in de novo Parkinson's disease patients.

BACKGROUND: Depigmentation of the substantia nigra (SN) and locus coeruleus (LC) is a conspicuous pathological feature of Parkinson's disease (PD) and is related to the loss of neuromelanin, whose paramagnetic properties result in high signal on specific T1-weighted magnetic resonance imaging (MRI). Recent studies have suggested that neuromelanin decrease in the SN and LC of PD patients may emerge as a possible diagnostic biomarker. The SN neuromelanin signal in de novo and early stage PD patients was studied to assess its diagnostic accuracy. This is the first study based on a semi-automated MRI analysis of the neuromelanin signal in de novo PD patients. METHODS: The inclusion criteria were untreated de novo PD and a 2-5 year disease duration; in addition, age matched healthy controls were enrolled. These were studied with a high-resolution T1-weighted MRI sequence at 3 T to visualize neuromelanin. The primary outcome was SN high signal area, length and neuromelanin/midbrain ratio obtained with semi-automated methods. RESULTS: A total of 12 de novo PD patients and 10 PD patients with a 2-5 year disease duration were evaluated. The area, length of the SN T1 high signal and the SN neuromelanin/midbrain ratio were markedly decreased in the PD groups compared with age-matched controls, with a substantial overlap between the two PD groups. CONCLUSIONS: Neuromelanin-sensitive MRI techniques can discriminate PD patients from healthy individuals with high sensitivity and specificity. Our findings are consistent with recent findings showing that PD neuromelanin changes remain stable during the course of the disease.

Stem cells and regenerative medicine in domestic and companion animals: a multispecies perspective.

Since their original isolation, the majority of the work on embryonic stem cells (ESC) has been carried out in mice. While the mouse is an outstanding model for basic research, it also has considerable limitations for translational work, especially in the area of regenerative medicine. This is due to a combination of factors that include physiological and size differences when compared to humans. In contrast, domestic animal species, such as swine, and companion animal species, such as dogs, provide unique opportunities to develop regenerative medicine protocols that can then be utilized in humans. Unfortunately, at present, the state of knowledge related to, and availability of, ESC from domestic animals vary among species such as pig, horse, dog and cat, and without exception lags significantly behind the mouse and human. It is clear that much still needs to be discovered. The 'stem cell-like' cell lines being reported are still not satisfactorily used in regenerative medicine, due to reasons such as heterogeneity and chromosomal instability. As a result, investigators have searched for alternate source of cells that can be used for regenerative medicine. This approach has uncovered a range of adult stem cells and adult progenitor cells that have utility in both human and veterinary medicine. Here, we review a range of stem cells, from ESC to induced pluripotent stem cells, and discuss their potential application in the field of regenerative medicine.

[Clinical characteristics and follow-up of 60 patients with recent diagnosis of giant cell arteritis, NEWTON study]. / Caractéristiques et profils évolutifs des patients avec un diagnostic récent d'artérite à cellules géantes, étude NEWTON.

INTRODUCTION: The management of giant cell arteritis (GCA) has evolved with the arrival of tocilizumab (TCZ) and the use of PET/CT. Our objective is to describe the characteristics and followup of patients with recent diagnosis of GCA in current care. PATIENTS AND METHODS: The NEWTON cohort is a monocentric retrospective cohort based on data collected from 60 GCA patients diagnosed between 2017 and 2022 according to the ACR/EULAR 2022 criteria. RESULTS: The median age at diagnosis was 73 [68.75; 81] years old. At diagnosis, the main manifestations were unusual temporal headaches in 48 (80 %) and an inflammatory syndrome in 50 (83 %) patients. Temporal artery biopsy confirmed the diagnosis in 49/58 (84 %) patients. Doppler of the temporal arteries found a halo in 12/23 (52 %) patients. The PET/CT found hypermetabolism in 19/43 (44 %) patients. Prednisone was stopped in 17.5 [12.75; 24.25] months. During follow-up, 22 (37 %) patients received TCZ. At least one complication of corticosteroid therapy was observed in 22 (37 %) patients. After a median follow-up of 24 [12; 42] months, 25 (42 %) patients relapsed. At the end of the follow-up, 29 (48.3 %) patients were weaned from corticosteroid therapy and 15 (25 %) were on TCZ. CONCLUSION: Despite the increasing use of TCZ in the therapeutic arsenal and of the PET/CT in the imaging tools of GCA patients, relapses and complications of corticosteroid therapy remain frequent, observed in more than a third of patients.

Canine fibroblasts expressing human transcription factors: what is in the route for the production of canine induced pluripotent stem cells.

The aim of this study was to further clarify the mechanisms involved in inducing pluripotency using canine foetal fibroblast cells. The two pluripotency-related transcription factors, OCT4 and SOX2, coupled to a fluorescent reporter gene were transduced, individually or in combination, using a lentiviral system. Stable transgenic cell lineages were obtained and canine cells showed to be highly responsive to the integration and expression of human SOX2 and OCT4, also depending on the amount of virus used for incubation. Such positive results are essential for the establishment of pluripotency induction through the incorporation of known transcription factors into the genome of somatic cells.

Transorbital Endoscopic Surgery for Sphenoid Wing Meningioma: Long-Term Outcomes and Surgical Technique.

Sphenoid wing meningiomas are benign tumors that can result in proptosis, visual impairment, and pain. Traditional open surgical approaches are associated with significant morbidity. Transorbital endoscopic surgery has been developed as a minimally invasive approach to gain access to these tumors and address the main presenting symptoms. Case series reporting transorbital endoscopic resection of sphenoid wing meningiomas using combined endonasal, pre-caruncular, and extended superior eyelid approaches have demonstrated stable and/or improved short- and medium-term visual outcomes. Earlier medial optic nerve decompression appears to result in more favorable long-term visual outcomes. Transorbital endoscopic surgery therefore represents an emerging minimally invasive alternative to deal with these challenging lesions.

Role of bacterial intimin in colonic hyperplasia and inflammation.

Enteropathogenic Escherichia coli (EPEC) cells adhere to gut epithelial cells through intimin alpha: the ligand for a bacterially derived epithelial transmembrane protein called the translocated intimin receptor. Citrobacter rodentium colonizes the mouse colon in a similar fashion and uses a different intimin: intimin beta. Intimin alpha was found to costimulate submitogenic signals through the T cell receptor. Dead intimin beta+ C. rodentium, intimin alpha-transfected C. rodentium or E. coli strain K12, and EPEC induced mucosal hyperplasia identical to that caused by C. rodentium live infection, as well as a massive T helper cell-type 1 immune response in the colonic mucosa. Mutation of cysteine-937 of intimin to alanine reduced costimulatory activity in vitro and prevented immunopathology in vivo. The mucosal changes elicited by C. rodentium were interferon-gamma-dependent. Immunopathology induced by intimin enables the bacteria to promote conditions that are favorable for increased microbial colonization.

'Smart' grommets.

OBJECTIVE: A novel, smartphone-based technique for endoscopic grommet insertion is presented.Results and conclusionThis method is both cost-effective and time-saving, offering a valuable alternative to the traditional microscope-based method in a resource-constrained setting.

Hearing outcomes in children with meningitis at Red Cross War Memorial Children's Hospital, Cape Town, South Africa: A silent crisis.

BACKGROUND: Bacterial meningitis is one of the most common causes of hearing loss. Hearing loss develops within the first few days following the onset of meningitis, highlighting the importance of early audiology referrals. Post-meningitis hearing loss may lead to cochlear ossification, making traditional cochlear implantation impossible. OBJECTIVES: To determine the duration of time from meningitis diagnosis to audiology referral. A second objective was to determine the prevalence and severity of meningitis-related sensorineural hearing loss (SNHL) at Red Cross War Memorial Children's Hospital (RCWMCH), a paediatric tertiary hospital in Cape Town, South Africa (SA). METHODS: A retrospective folder review of all children diagnosed with meningitis and referred for audiological evaluation was conducted over an 18-month period between 1 January 2015 and 30 June 2016. Descriptive statistical methods were used for data analysis. RESULTS: The total number of inpatients diagnosed with unspecified meningitis at RCWMCH between January 2015 and June 2016 was 345. The total number diagnosed with bacterial meningitis was 68. Only 16 children with meningitis (23.5%) were referred to the Department of Audiology at RCWMCH over the 18-month period. Twelve children had confirmed bacterial meningitis. All the children had up-to-date immunisations. The mean (standard deviation) time from diagnosis to audiology referral was 17 weeks (16.9; range 1 - 60). The overall prevalence of hearing loss was 42.8%. Four children (28.5%) were diagnosed with severe to profound SNHL. CONCLUSIONS: Less than a quarter of all children diagnosed with bacterial meningitis at RCWMCH over the 18-month study period were referred to audiology. Frequent delays in referrals for audiological assessment following meningitis were noted. This may indicate a lack of awareness on the part of doctors. Healthcare practitioners need to be made aware of the need for prompt audiological testing for children with meningitis. In SA, local guidelines for early diagnosis and monitoring of hearing in children who have had meningitis should be developed in order to improve hearing outcomes.

Liver Transplantation Using Prolonged Cold Ischemia Time Grafts Preserved With Institute George-Lopez-1 Solution.

Institute George-Lopez-1 (IGL-1) solution is a preservation solution with lower potassium and lower viscosity than the University of Wisconsin solution that has been recently used in liver transplantation. In the present series, we compare the outcome of liver grafts from brain-dead donors preserved in IGL-1cold storage solution, with cold ischemia times (CITs) longer than 8 hours and those less than 8 hours. Two hundred fifty-two liver transplantations performed from January 2014 to December 2016 at Hospital Santa Isabel, Blumenau, Brazil, were retrospectively analyzed. The patients were divided in two groups according to the CIT. Group I patients (N = 155) had less than 8 hours of CIT with a mean age of 54 ± 11.35 years, whereas group II patients (N = 97) had more than 8 hours of CIT with a mean age of 52 ± 12.5 years. There was no difference between the groups related to indication for liver transplantation and donor characteristics. The only difference statically significant on laboratory data was between the levels of aspartate aminotransferase at day 1 after transplantation. On day 7 post-transplantation there was no difference statistically significant between aspartate aminotransferase, alanine aminotransferase, and bilirubin levels between the two groups. Similar 1-year patient survival rates were found in both groups, with 85.88% for group I and 85.75% in group II. The IGL-1 solution has been shown to be safe, effective, and with good results in liver transplantations. Early graft function and 1-year patient survival rates did not differ when grafts preserved for less than 8 hours were compared to those with CIT greater than 8 hours.

Gastroprotective and anti-inflammatory activities integrated to chemical composition of Myracrodruon urundeuva Allemão - A conservationist proposal for the species.

ETHNOPHARMACOLOGICAL RELEVANCE: Myracrodruon urundeuva Allemão (Aroeira-do-Sertão), Anacardiaceae, is one of the most used plants in folk medicine in Northeastern Brazil as an anti-inflammatory, healing and antiulcer. This species is threatened with extinction due to anthropogenic exploitation. The importance of this study is to demonstrate the feasibility of a conservationist model of replacement of the M. urundeuva adult tree (inner bark) for its under developing plants (shoots) in order to ensure the preservation of this species, but also to ensure sufficient raw material for pharmaceutical purposes. AIM OF THE STUDY: To characterize chemically and assess the gastroprotective and anti-inflammatory activities of the fluid extracts from M. urundeuva innebark (adult plant) as well as stem and leaves of shoots (young plant). MATERIALS AND METHODS: The fluid extracts were prepared by maceration-percolation with hydroalcoholic solution according to the methodology described in the Brazilian Pharmacopoeia. These extracts were cleaned-up through solid phase extraction (SPE) and chemically characterized by ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-ESI-QTOF MS/MS). Gastroprotective and anti-inflammatory activities of the extracts (700 or 1000 mg/kg) were assessed on ethanol-induced gastric lesions and Croton oil-induced ear edema in rats, respectively. The extracts were evaluated for cytotoxicity in vitro. RESULTS: The UPLC-ESI-QTOF-MS/MS analysis evidenced the presence of chalcones, flavonoids and tannins. Gastroprotective and anti-inflammatory activities achieved with fluid extracts from the stems and leaves was similar to inner bark. The fluid extracts were not toxic. CONCLUSION: It is possible to replace the inner bark of the adult tree for the stems and leaves from the shoots as raw material to be used in the preparation of its the phytotherapeutics. Therefore, this finding may help in the implementation of public policies that ensure the conservation of the species along with its sustainable use for pharmaceutical purposes.

Generation of LIF-independent induced pluripotent stem cells from canine fetal fibroblasts.

Takahashi and Yamanaka established the first technique in which transcription factors related to pluripotency are incorporated into the genome of somatic cells to enable reprogramming of these cells. The expression of these transcription factors enables a differentiated somatic cell to reverse its phenotype to an embryonic state, generating induced pluripotent stem cells (iPSCs). iPSCs from canine fetal fibroblasts were produced through lentiviral polycistronic human and mouse vectors (hOSKM/mOSKM), aiming to obtain pluripotent stem cells with similar features to embryonic stem cells (ESC) in this animal model. The cell lines obtained in this study were independent of LIF or any other supplemental inhibitors, resistant to enzymatic procedure (TrypLE Express Enzyme), and dependent on bFGF. Clonal lines were obtained from slightly different protocols with maximum reprogramming efficiency of 0.001%. All colonies were positive for alkaline phosphatase, embryoid body formation, and spontaneous differentiation and expressed high levels of endogenous OCT4 and SOX2. Canine iPSCs developed tumors at 120 days post-injection in vivo. Preliminary chromosomal evaluations were performed by FISH hybridization, revealing no chromosomal abnormality. To the best of our knowledge, this report is the first to describe the ability to reprogram canine somatic cells via lentiviral vectors without supplementation and with resistance to enzymatic action, thereby demonstrating the pluripotency of these cell lines.

Effect of xylitol:sorbitol on fluoride enamel demineralization reduction in situ.

OBJECTIVES: To evaluate if sugar alcohols would reduce enamel demineralization enhancing the fluoride (F) effect. METHODS: A crossover in situ study was conducted in four phases, during which 10 volunteers were submitted to one of the treatments: (I) Distilled and deionized water, as a negative control; (II) F (226 microg F/ml as NaF; concentration used in commercial mouthrinse); (III) X:S (xylitol:sorbitol 1:3; final concentration 1.6M; 28% of sugar alcohols) and (IV) F+X:S (same final concentration that groups II and III). The volunteers wore palatal appliances containing four bovine enamel blocks of known surface microhardness (SMH), covered with a 'test plaque' of mutans streptococci, which were immersed during 1 min in one of the allocated treatment solutions simultaneously that the volunteers rinsed their mouths with the same solution. After the rinsing the appliances were put in the mouth and after 20 min a cariogenic challenge was made with 20% sucrose solution during 1 min. After further 45 min the 'test plaque' was collected for F analysis, enamel SMH was again determined and the percentage of change in relation to baseline was calculated; F uptake in enamel was also determined. RESULTS: With respect to all the analyses made, the group F+X:S did not differ from the F treatment (p>0.05) and the groups treated with F and F+X:S differed from the negative control (p<0.05). CONCLUSIONS: The results suggest that xylitol:sorbitol may not enhance the effect of fluoride present in mouth rinse on the reduction of enamel demineralization.