Exposure of humans to the zoonotic nematode Dirofilaria immitis in Northern Portugal.

Dirofilariosis caused by Dirofilaria immitis (heartworm) is a zoonosis, considered an endemic disease of dogs and cats in several countries of Western Europe, including Portugal. This study assesses the levels of D. immitis exposure in humans from Northern Portugal, to which end, 668 inhabitants of several districts belonging to two different climate areas (Csa: Bragança, Vila Real and Csb: Aveiro, Braga, Porto, Viseu) were tested for anti-D. immitis and anti-Wolbachia surface proteins (WSP) antibodies. The overall prevalence of seropositivity to both anti-D. immitis and WSP antibodies was 6.1%, which demonstrated the risk of infection with D. immitis in humans living in Northern Portugal. This study, carried out in a Western European country, contributes to the characterisation of the risk of infection with D. immitis among human population in this region of the continent. From a One Health point of view, the results of the current work also support the close relationship between dogs and people as a risk factor for human infection.

Clinical study on the efficacy of LED phototherapy for pain control in an orthodontic procedure.

Pain is an unpleasant and emotional subjective sensory experience that occurs during orthodontic procedures. Currently, LED phototherapy is an alternative to the use of laser light as analgesic agent due to similarity of response and lower cost. This case-control, quantitative, qualitative, and longitudinal study aimed to investigate the effect of IR LED phototherapy (λ846 ± 20 nm) in pain during the process of tooth separation during orthodontic treatment. After approval by the Institution Ethics Committee, 40 patients (30 female/10 male, 20-30 years old, average age 24.5 ± 2.6 years old) fulfilling the inclusion criteria entered the study and received a set of four visual analog scales (VAS) for scoring pain immediately, 48 h, 72 h, and 7 days after the insertion of the separating elastics. The patients were randomly distributed into two groups (experimental and control). The patients of experimental group received LED phototherapy (180 mW, 22 s, 4 J, 8 J/cm2, 0.36 W/cm2, spot of 0.5 cm2, spot diameter 0.8 cm) at the same times in which VAS was performed, and control patients were not irradiated. It was found that, in both groups, there was an increase in pain 48 h after insertion of the elastic tooth separator, decreasing 72 h after its installation and reached the lowest level of pain after 7 days. Comparison between groups showed that pain level in the LED group was always statistically significantly lower (p < 0.05), except for the time of installation (T1). The use of LED light was effective in significantly reducing the level of pain after insertion of the elastic tooth separators when compared to the control group.

Alzheimer's disease-related amyloid-ß1-42 peptide induces the loss of human sperm function.

Characteristically identified as the main component of senile plaques present in patients suffering from Alzheimer's disease, Aß has been detected in human testis and reproductive fluids, but its effect on spermatozoa has not been addressed. The present study evaluated whether the most toxic and aggregant amyloid precursor protein (APP)-proteolytic product, amyloid-ß1-42 (Aß1-42), was capable of affecting sperm functionality. Normozoospermic samples were either exposed to different Aß1-42 doses or to the untreated and scrambled controls for a maximum of 48 h at 37 °C and 5%CO2, and motility, viability and mitochondrial status were evaluated. Additionally, tyrosine phosphorylation was analyzed by immunocytochemistry and acrosomal integrity through PSA-FITC. A shorter treatment period was used to monitor prompt Ca2+ responses. Aß1-42 peptide decreased motility before inducing mitochondrial impairment (p < 0.05; n = 6). Both outcomes became more pronounced with time, reaching their maximal decrease at 48 h, where even 1 µM produced undesirable effects (p < 0.05; n = 6). Aß1-42 peptide also decreased cell survival (p < 0.05; n = 6). Furthermore, although no effects on tyrosine phosphorylation were observed (p > 0.05; n = 6), reduced acrosomal integrity was detected (p < 0.05; n = 7), which was not correlated with viability loss (p > 0.05). In parallel, all Aß1-42 concentrations elicited a [Ca2+]i rise but a significant difference was only observed at 20 µM (p < 0.05; n = 7) and a tendency was obtained with 10 µM (p = 0.053; n = 7). In conclusion, Aß1-42 peptide oligomers impair sperm function in vitro, although further studies are required to determine the clinical relevance of these findings.

Seroprevalence of heartworm (Dirofilaria immitis) in feline and canine hosts from central and northern Portugal.

Dirofilaria immitis is endemic in Portugal. Several studies have reported the presence of canine heartworm disease, although no previous studies on feline infections have been published. The aim of this study was to determine the prevalence of D. immitis in cats and dogs from central and northern Portugal. Blood samples from 434 cats were tested for circulating anti-D. immitis and anti-Wolbachia antibodies. Furthermore, 386 dogs were tested for circulating D. immitis antigens. Overall feline seroprevalence was 15%, while canine prevalence was 2.1%. The highest feline seroprevalences of 18.7% and 17.6% were found in Aveiro and Viseu, respectively, while the highest canine prevalences of 8.8% and 6.8% were found in Coimbra and Aveiro, respectively. Cats and dogs showing respiratory signs presented higher prevalences of 24.4% and 17%, respectively, while 50% of cats with gastrointestinal signs were seropositive. The present study confirms the seropositivity of D. immitis in the feline population in central and northern Portugal, and suggests the importance of including heartworm disease in the list of differential diagnoses of cats and dogs showing clinical signs compatible with the disease.

Antifungal activity of terpenes isolated from the Brazilian Caatinga: a review.

Terpenoids, also named terpenes or isoprenoids, are a family of natural products found in all living organisms. Many plants produce terpenoids as secondary metabolites, and these make up a large part of essential oils. One of most important characteristic is that the compounds are volatile, have odor and can be used in a variety of applications in different industrial segments and traditional medicine. Brazil has a rich and diverse flora that can be used as a source of research for obtaining new molecules. Within the Brazilian flora, it is worth mentioning the Caatinga as an exclusively Brazilian biome where plants adapt to a specific series of weather conditions and therefore become a great storehouse of the terpenoid compounds to be described herein. Fungal infections have become increasingly common, and a great demand for new agents with low toxicity and side effects has thus emerged. Scientists must search for new molecules exhibiting antifungal activity to develop new drugs. This review aims to analyze scientific data from the principal published studies describing the use of terpenes and their biological applications as antifungals.

Analysis of the toxicological and pharmacokinetic profile of Kaempferol-3-O-ß-D-(6"-E-p-coumaryl) glucopyranoside - Tiliroside: in silico, in vitro and ex vivo assay.

Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as "pitó". This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O-ß-D-(6"-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential.

In silico, in vitro, and ex vivo studies of the toxicological and pharmacological properties of the flavonoid 5,7-dihydroxy-3,8,4'-trimethoxy.

Phytochemical studies of the species Pavonia glazioviana were performed. Quercetin, kaempferol, acacetin, and trimethoxylated flavonoid compounds (which present biological activity) were isolated. We aimed to evaluate the in silico, in vitro, and ex vivo toxicity of flavonoid 5,7-dihydroxy-3,8,4'-trimethoxy (Pg-1) obtained from P. glazioviana through chemical structure analyses, toxicity assessment, and predictive bioactive properties, using human samples in in vitro tests. In silico analysis suggested that Pg-1 presents a good absorption index for penetrating biological membranes (for oral bioavailability), while also suggesting potential antimutagenic, anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic, and apoptosis agonist bioactivities. Assessment of hemolytic and genotoxic effects revealed low hemolysis rates in red blood cells with no cellular toxicity in oral mucosa cells. The reduced cytotoxic activity suggested the safety of the concentrations used (500-1000 µg/mL), and demonstrated the varied interactions of Pg-1 with the analyzed cells. The data obtained in the present study suggested potential therapeutic application, and the non-toxic profile indicated viability for future studies.

Prognostic value of odor identification impairment in multiple sclerosis: 10-Years follow-up.

BACKGROUND: Olfactory dysfunction has been linked to clinical severity variables in multiple MS populations. Though, its prognostic value is still unknown. OBJECTIVE: The aim of this study was to explore the long-term outcome associated with Brief-Smell Identification Test (B-SIT) performance in a cohort of MS patients. METHODS: A retrospective review of the clinical records was conducted in 149 patients who participated in a previous study, with a median follow-up of 121 months. Demographic and clinical data regarding the last clinical appointment with EDSS measurement were collected. Multiple Sclerosis Severity Scale (MSSS) and Age-Related Multiple Sclerosis Severity (ARMSS) scores were calculated. Date of the last clinical contact or death was recorded. RESULTS: Among MS patients with progressive clinical course (n = 33), those with impaired B-SIT at baseline had greater change per month during follow-up (as measured by increases in MSSS and ARMSS scores) and a higher hazard of death. No significant associations were found among patients with relapsing and remitting MS (n = 116). CONCLUSIONS: The study results demonstrate that odor identification impairment has prognostic value in progressive MS, suggesting that a brief odor identification measure can be a marker of neurodegeneration in progressive MS.

Urotensin II-induced increase in myocardial distensibility is modulated by angiotensin II and endothelin-1.

Endogenous regulators, such as angiotensin-II (AngII), endothelin-1 (ET-1) and urotensin-II (U-II) are released from various cell types and their plasma levels are elevated in several cardiovascular diseases. The present study evaluated a potential crosstalk between these systems by investigating if the myocardial effects of U-II are modulated by AngII or ET-1. Effects of U-II (10(-8), 10(-7), 10(-6) M) were tested in rabbit papillary muscles in the absence and in the presence of losartan (selective AT(1) receptor antagonist), PD-145065 (nonselective ET-1 receptors antagonist), losartan plus PD-145065, AngII or ET-1. U-II promoted concentration-dependent negative inotropic and lusitropic effects that were abolished in all experimental conditions. Also, U-II increased resting muscle length up to 1.008+/-0.002 L/L(max). Correcting it to its initial value resulted in a 19.5+/-3.5 % decrease of resting tension, indicating increased muscle distensibility. This effect on muscle length was completely abolished in the presence of losartan and significantly attenuated by PD-145065 or losartan plus PD-145065. This effect was increased in the presence of AngII, resulting in a 27.5+/-3.9 % decrease of resting tension, but was unaffected by the presence of ET-1. This study demonstrated an interaction of the U-II system with the AngII and ET-1 systems in terms of regulation of systolic and diastolic function.

Nitric oxide and prostaglandins - important players in endothelin-1 induced myocardial distensibility.

This study investigated whether endothelin (ET)-1-induced increase in myocardial distensibility is preserved in heart failure (HF) and whether it is modulated by nitric oxide (NO) and prostaglandins. New Zealand white rabbits were treated with doxorubicin (1 mg/kg, intravenously twice a week for 8 weeks, DOX-HF group) or saline (control group). Effects of ET-1 (0.1, 1, 10 nM) were tested in papillary muscles from the DOX-HF group and a control group in the presence of: i) intact endocardial endothelium (EE); ii) damaged EE; iii) N(G)-nitro-L-arginine (L-NNA; NO synthase inhibitor), and iv) indomethacin (INDO; cyclooxygenase inhibitor). In the presence of an intact EE, ET-1 promoted concentration-dependent positive inotropic and lusitropic effects that were maintained after damaging the EE, in the presence of L-NNA or INDO and in the DOX-HF Group. ET-1 reduced resting tension at the end of the isometric twitch (increased diastolic distensibility) by 3.2+/-1.3 %, 6.0+/-1.6 % and 8.8+/-2.7 % (at 0.1, 1 and 10 nM, respectively), in muscles with intact EE, effect that was completely abolished after damaging EE, in the presence of L-NNA or INDO or in the DOX-HF Group. This study demonstrated that the increase in myocardial distensibility induced by ET-1 is absent in HF and is dependent of NO and prostaglandin release.

Analysis of the toxicological and pharmacokinetic profile of Kaempferol-3-O-ß-D-(6"-E-p-coumaryl) glucopyranoside - Tiliroside: in silico, in vitro and ex vivo assay / Análise do perfil toxicológico e farmacocinético de Kaempferol-3-O-ß-D- (6 "-E-p-cumaril) glucopiranosídeo - Tilirosídeo: ensaio in silico, in vitro e ex vivo

Abstract Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as "pitó". This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O-β-D-(6"-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential.

Resumo O tilirosídeo é um flavonóide glicosídico presente em muitas espécies de plantas, incluindo Helicteres velutina K. Schum (Malvaceae sensu lato), conhecida no Brasil como "pitó". Esta molécula mostrou ter muitas atividades biológicas, porém nenhum estudo foi realizado para investigar a toxicidade dessa substância. O presente trabalho teve como objetivo avaliar a possível toxicidade celular in silico, in vitro e ex-vivo do kaempferol-3-O-β-D- (6 "-Ep-coumaroil) glucopiranosídeo (tilirosídeo), por meio de análises de estrutura química, toxicidade avaliação e propriedades bioativas preditivas, utilizando amostras humanas para testes in vitro e ex-vivo. A análise in silico sugere que o tilirosídeo exibe bom índice de absorção para penetrar nas membranas biológicas. Além disso, apresentou considerável potencial de proteção celular contra os radicais livres e com atividades anticarcinogênica, antioxidante, antineoplásica, antiinflamatória, anti-hemorrágica e antitrombótica. A avaliação dos efeitos hemolíticos e genotóxicos do tilirosídeo mostrou baixas taxas de hemólise nas hemácias e ausência de toxicidade em células da mucosa oral. Os dados obtidos indicam que esta molécula pode possuir uma abordagem terapêutica promissora como uma possível nova droga com potencial biotecnológico.

Analysis of the toxicological and pharmacokinetic profile of Kaempferol-3-O-β-D-(6"-E-p-coumaryl) glucopyranoside - Tiliroside: in silico, in vitro and ex vivo assay / Análise do perfil toxicológico e farmacocinético de Kaempferol-3-O-β D-(6"-E-p-cumaril) glucopiranosídeo - Tilirosídeo: ensaio in silico, in vitro e ex vivo

Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as "pitó". This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O-β-D-(6”-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential.

O tilirosídeo é um flavonóide glicosídico presente em muitas espécies de plantas, incluindo Helicteres velutina K. Schum (Malvaceae sensu lato), conhecida no Brasil como “pitó”. Esta molécula mostrou ter muitas atividades biológicas, porém nenhum estudo foi realizado para investigar a toxicidade dessa substância. O presente trabalho teve como objetivo avaliar a possível toxicidade celular in silico, in vitro e ex-vivo do kaempferol-3-O-β-D- (6 ”-Ep-coumaroil) glucopiranosídeo (tilirosídeo), por meio de análises de estrutura química, toxicidade avaliação e propriedades bioativas preditivas, utilizando amostras humanas para testes in vitro e ex-vivo. A análise in silico sugere que o tilirosídeo exibe bom índice de absorção para penetrar nas membranas biológicas. Além disso, apresentou considerável potencial de proteção celular contra os radicais livres e com atividades anticarcinogênica, antioxidante, antineoplásica, antiinflamatória, anti-hemorrágica e antitrombótica. A avaliação dos efeitos hemolíticos e genotóxicos do tilirosídeo mostrou baixas taxas de hemólise nas hemácias e ausência de toxicidade em células da mucosa oral. Os dados obtidos indicam que esta molécula pode possuir uma abordagem terapêutica promissora como uma possível nova droga com potencial biotecnológico.

Analysis of the toxicological and pharmacokinetic profile of Kaempferol-3-O--D-(6-E-p-coumaryl) glucopyranoside - Tiliroside: in silico, in vitro and ex vivo assay

Abstract Tiliroside is a glycosidic flavonoid present in many plants species including Helicteres velutina K. Schum (Malvaceae sensu lato), commonly known in Brazil as pitó. This molecule has been shown to have many biological activities, however no study has been carried out to investigate the toxicity of this substance. The present work aimed to evaluate the possible cellular toxicity in silico, in vitro and ex-vivo of the kaempferol-3-O--D-(6-E-p-coumaroyl) glucopyranoside (tiliroside), through chemical structure analysis, toxicity assessment and predictive bioactive properties, using human samples for in vitro and ex-vivo tests. The in silico analysis suggests that tiliroside exhibited great absorption index when penetrating biological membranes. In addition, it also displayed considerable potential for cellular protection against free radicals, and anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic and antithrombotic activities. The assessment of the hemolytic and genotoxic effects of tiliroside showed low hemolysis rates in red blood cells and absence of cellular toxicity in the oral mucosa cells. The data obtained indicate that this molecule could be a promising therapeutic approach as a possible new drug with biotechnological potential.

Resumo O tilirosídeo é um flavonóide glicosídico presente em muitas espécies de plantas, incluindo Helicteres velutina K. Schum (Malvaceae sensu lato), conhecida no Brasil como pitó. Esta molécula mostrou ter muitas atividades biológicas, porém nenhum estudo foi realizado para investigar a toxicidade dessa substância. O presente trabalho teve como objetivo avaliar a possível toxicidade celular in silico, in vitro e ex-vivo do kaempferol-3-O--D- (6 -Ep-coumaroil) glucopiranosídeo (tilirosídeo), por meio de análises de estrutura química, toxicidade avaliação e propriedades bioativas preditivas, utilizando amostras humanas para testes in vitro e ex-vivo. A análise in silico sugere que o tilirosídeo exibe bom índice de absorção para penetrar nas membranas biológicas. Além disso, apresentou considerável potencial de proteção celular contra os radicais livres e com atividades anticarcinogênica, antioxidante, antineoplásica, antiinflamatória, anti-hemorrágica e antitrombótica. A avaliação dos efeitos hemolíticos e genotóxicos do tilirosídeo mostrou baixas taxas de hemólise nas hemácias e ausência de toxicidade em células da mucosa oral. Os dados obtidos indicam que esta molécula pode possuir uma abordagem terapêutica promissora como uma possível nova droga com potencial biotecnológico.

In silico, in vitro, and ex vivo studies of the toxicological and pharmacological properties of the flavonoid 5,7-dihydroxy-3,8,4'-trimethoxy

Phytochemical studies of the species Pavonia glazioviana were performed. Quercetin, kaempferol, acacetin, and trimethoxylated flavonoid compounds (which present biological activity) were isolated. We aimed to evaluate the in silico, in vitro, and ex vivo toxicity of flavonoid 5,7-dihydroxy-3,8,4'-trimethoxy (Pg-1) obtained from P. glazioviana through chemical structure analyses, toxicity assessment, and predictive bioactive properties, using human samples in in vitro tests. In silico analysis suggested that Pg-1 presents a good absorption index for penetrating biological membranes (for oral bioavailability), while also suggesting potential antimutagenic, anticarcinogenic, antioxidant, antineoplastic, anti-inflammatory, anti-hemorrhagic, and apoptosis agonist bioactivities. Assessment of hemolytic and genotoxic effects revealed low hemolysis rates in red blood cells with no cellular toxicity in oral mucosa cells. The reduced cytotoxic activity suggested the safety of the concentrations used (500-1000 µg/mL), and demonstrated the varied interactions of Pg-1 with the analyzed cells. The data obtained in the present study suggested potential therapeutic application, and the non-toxic profile indicated viability for future studies.

The content of the LRS chamber provides a new quality tool for the characterization of the donor platelet profile.

Cobe Trima version 4 is an apheresis system designed for the collection of combinations of Red Blood Cells (RBC), Platelets (PLT) and plasma components from a single donor. The validation of this apheresis system for PLT components in our institution evidenced the LRS efficacy, as leucoreduction was attained on a 100% basis. However, there were some unexpected occurrences regarding the PLT content. Certain donations showed large disagreements between programmed and obtained yields, not being found clear reasons for those outcomes. Furthermore, the mean platelet volume (MPV) was relatively low when compared with other platelets components produced by other methodologies. An investigation was initiated in order to know whether these shortcomings were donor or process/instrument related. A link was established between the raw material (donor), the process/instrument and the final product, and a new tool was introduced by the study of the LRS chamber. The LRS chamber content was assessed and the PLT cellular indices and PLT aggregation states compared with those obtained from the respective donor and the final product from the same origin. The storage stability of the final products was also analyzed based on these same tests to investigate if the initial low MPV had any deleterious effect during the shelf life of the components. Twenty five plateletpheresis donors, three of them new ones, were randomly selected for this study. For the first time, the content of the LRS chamber was used as a quality tool for identification of the cause of unexpected yields. Large aggregates remained in the LRS chamber in certain donors who were prone to undergo spontaneous aggregation, making the platelet yields low and platelet MPV very small. Based on repeated failures and on this new quality parameter showing LRS chamber abnormality, two donors were temporarily deferred from the panel. While it was not possible to identify the causes, it is appropriate to raise the question whether these profiles were Trima version 4 specific or not. Hence, further investigation is needed for a better clarification. In short, the simultaneous analysis of products and LRS content provided useful information not only for the characterization of donor-related phenomena but also helped in the identification of potential shortcomings in the machine performance allowing for remedial action to be taken on evidence based data.

Autologous hematopoietic SCT normalizes miR-16, -155 and -142-3p expression in multiple sclerosis patients.

Autologous hematopoietic SCT (AHSCT) has been investigated in the past as a therapeutic alternative for multiple sclerosis (MS). Despite advances in clinical management, knowledge about mechanisms involved with clinical remission post transplantation is still limited. Abnormal microRNA and gene expression patterns were described in MS and have been suggested as disease biomarkers and potential therapeutic targets. Here we assessed T- and B-cell reconstitution, microRNAs and immunoregulatory gene expression after AHSCT. Early immune reconstitution was mainly driven by peripheral homeostatic proliferation. AHSCT increased CD4(+)CD25(hi)FoxP3(+) regulatory T-cell counts and expression of CTLA-4 and GITR (glucocorticoid-induced TNFR) on CD4(+)CD25(hi) T cells. We found transient increase in exhausted PD-1(+) T cells and of suppressive CD8(+)CD28(-)CD57(+) T cells. At baseline, CD4(+) and CD8(+) T cells from MS patients presented upregulated miR-16, miR-155 and miR-142-3p and downregulated FOXP3, FOXO1, PDCD1 and IRF2BP2. After transplantation, the expression of FOXP3, FOXO1, PDCD1 and IRF2BP2 increased, reaching control levels at 2 years. Expression of miR-16, miR-155 and miR-142-3p decreased towards normal levels at 6 months post therapy, remaining downregulated until the end of follow-up. These data strongly suggest that AHSCT normalizes microRNA and gene expression, thereby improving the immunoregulatory network. These mechanisms may be important for disease control in the early periods after AHSCT.

Visual topography of V1 in the Cebus monkey.

The representation of the visual field in the striate cortex (V1) was mapped with multiunit electrodes in the Cebus monkey. Nine Cebus apella, anesthetized with N2O and immobilized with pancuromium bromide were studied in repeated recording sessions. In each hemisphere, V1 contains a continuous representation of the contralateral visual hemifield. The representation of the vertical meridian (VM) forms the external border of V1 except at the anteriormost portion of the calcarine fissure. The representation of the horizontal meridian (HM) divides the area so that the representation of the lower visual field is located dorsally, and that of the upper field ventrally. The convoluted surface of V1 can be only partially unfolded, and no precise "flattened" map can be obtained without introducing surface discontinuities. The visual topography of V1 is presented in a series of coronal sections and in "flattened" maps. The representation of the central visual field is magnified relative to that of the periphery in V1. The evaluation of the cortical magnification factors measured along isoeccentric and isopolar dimensions in the partially unfolded model of V1 revealed anisotropies in the representation of the visual field with larger magnification along isopolar lines than along isoeccentric lines. Receptive field size increases with increasing eccentricity, whereas point image size decreases with increasing eccentricity.

Representation of the visual field in the second visual area in the Cebus monkey.

The representation of the visual field in the second visual area (V2) was reconstructed from multiunit visual responses and anatomical tracers. Receptive field plotting was performed during multiple recording sessions in seven Cebus apella monkeys under N2O/O2 and immobilized with pancuronium bromide. V2 forms a continuous belt of variable width around striate cortex (V1) except at the most anterior portion of the calcarine sulcus. In each hemisphere V2 contains a visuotopic representation of the contralateral visual hemifield. The representation of the vertical meridian is adjacent to that of V1 and forms the posterior border of V2. The representation of the fovea of V2 is adjacent to that of V1. The representation of the horizontal meridian (HM) is continuous with that of V1; then it splits to form the anterior border of V2, both dorsally and ventrally. The lower quadrant of the visual field is represented dorsally and the upper quadrant ventrally. The visual topography of V2 is coarser than that of V1. In V2, receptive fields corresponding to recording sites separated by a cortical distance of up to 4 mm may represent the same portion of the visual field. In three additional animals, combined injections of fluorescent tracers along the HM representation in V1 yielded two projection sites at the anterior border of V2. The split of the HM representation is estimated to occur at an eccentricity below 1 degree. Quantitative analysis showed that in V2 the representation of the central visual field is magnified relative to that of the periphery. The cortical magnification factor is greater along the isopolar dimension than along the isoeccentric one. Receptive field size in V2 increases with increasing eccentricity. In sections stained for myelin by the Heidenhein-Wöelcke method V2 can be distinguished from the surrounding cortex for most of its extent.

Topographic organization of cortical input to striate cortex in the Cebus monkey: a fluorescent tracer study.

Cortical afferents to area V1 were studied in seven Cebus monkeys by means of retrograde fluorescent tracers. Injections were placed in V1, under electrophysiological guidance, in the regions of representation of both the upper and lower visual quadrants, at eccentricities that ranged from 0.5 to 64 degrees. In all cases retrogradely filled neurons were found in retinotopically corresponding portions of areas V2 and MT, as defined electrophysiologically (Rosa et al: J. Comp. Neurol. 275:326, 1988; Fiorani et al: J Comp Neurol 287:98, 1989). The results also revealed two other visual zones located anterior to V2 here named third and fourth visual areas. A topographical organization of the connections was observed in these areas, with upper quadrant located ventrally and lower quadrant located dorsally. A clear central-peripheral gradient, from the lateral to the medial cortical surface, was also observed in these areas. Lower field injections revealed crude topographic organization in area DZ and a diffuse projecting zone in the annectent gyrus. Peripheral injections in V1 revealed a clear upper and lower field segregation in areas PO and prostriata as well as a complex topography in MST. In addition, another region of labeling revealed the presence of an area, the temporal ventral posterior region, with an organized topographic representation of the upper field, with a central to peripheral gradient, from the lateral to the medial cortical surface. Three groups of cortical areas were distinguished according to the laminar distribution of neurons labeled from V1. In the first group, which is characterized by dense infra- and supragranular labeling, only V2 was included. The second group consists of areas V3, MT, and PO. These areas show dense labeling in the infragranular layers and occasionally sparse labeling in the supragranular layers. Finally, V4 and the other projecting areas, which are characterized by exclusive labeling of the infragranular layers were included in the third group.