Urinary sodium, iodine, and volume in relation to metabolic syndrome in Mesoamerican children and their parents.

BACKGROUND AND AIMS: The roles of sodium or iodine intake on the metabolic syndrome (MetS) etiology remain controversial. We evaluated the associations of 24 h urinary sodium and iodine with MetS among Mesoamerican children and their adult parents. METHODS AND RESULTS: We conducted a cross-sectional study among 217 school-age children and 478 parents from 9 Mesoamerican cities. Exposures were high 24 h urinary sodium excretion and concentration (>2000 mg/d or mg/L, respectively) and high 24 h urinary iodine excretion and concentration (≥300 µg/d or µg/L, respectively). In children, the outcome was a standardized metabolic score from five criteria analogous to the Adult Treatment Panel (ATP) III criteria. In adults, MetS was defined according to the ATP III criteria. We estimated adjusted mean differences in the metabolic risk score and adjusted prevalence ratios of MetS between exposure categories using multivariable regression. In children, high sodium concentration was associated with a 0.10 units (43% of a SD) higher score (P = 0.001) and high iodine concentration was related to a 0.09 units (39% of a SD) higher score (P = 0.009). Unexpectedly, high 24 h urinary volume was associated with a lower metabolic score. In adults, high 24 h sodium excretion was related to hypertension and high iodine concentration was related to increased MetS prevalence. CONCLUSION: High sodium and iodine concentrations, but not 24 h iodine excretion, are significantly associated with MetS in children, whereas high 24 h urinary volume is related to a decreased metabolic score. In adults, high iodine concentration tends to be related to increased MetS prevalence, but not 24 h iodine excretion.

Fibroblast growth factor-2 promotes in vitro activation of cat primordial follicles.

This study evaluated the effect of fibroblast growth factor-2 (FGF-2) on the morphology, primordial follicle activation and growth after in vitro culture of domestic cat ovarian tissue. Ovaries (n = 12) from prepubertal domestic cats were collected and fragmented. One fragment was fixed for histological analysis (fresh control). The remaining fragments were incubated in control medium alone or with 10, 50 or 100 ng/ml FGF-2 for 7 days. After in vitro culture, the following endpoints were analyzed: morphology, activation by counting primordial and developing follicles, and growth (follicle and oocyte diameters). Treatment with 100 ng/ml FGF-2 maintained (P > 0.05) the percentage of normal follicles similar to fresh control. Follicle survival was greater (P < 0.05) after culture in 100 ng/ml FGF-2 than in 50 ng/ml FGF-2. The percentage of primordial follicles decreased (P < 0.05) and the percentage of developing follicles increased (P < 0.05) in all treatments compared with fresh tissue. The proportion of developing follicles increased (P < 0.05) in tissues incubated with 100 ng/ml FGF-2 compared with control medium and other FGF-2 concentrations. Furthermore, culture in 10 or 100 ng/ml FGF-2 resulted in increased (P < 0.05) follicle and oocyte diameters compared with fresh tissues and MEM+. In conclusion, FGF-2 at 100 ng/ml maintains follicle survival and promotes the in vitro activation and growth of cat primordial follicles.

Achados clínicos, histopatológicos e moleculares da mielopatia degenerativa canina: relato de caso / Clinical, histopathological and molecular findings of canine degenerative myelopathy: case report

Objetivou-se descrever os achados clínicos, histopatológicos e moleculares associados à MDC em um cão da raça Pastor-Suiço. O cão possuía uma paraparesia progressiva em membros pélvicos e foi submetido a avaliações clínicas, pelas quais se obteve, entre outros diferenciais, o diagnóstico presuntivo de MDC. Com a evolução dos sinais, o tutor optou pela eutanásia. Os achados histopatológicos da medula espinhal foram compatíveis com uma degeneração segmentar axonal e mielínica. O diagnóstico molecular foi realizado por meio da extração do DNA obtido por swab oral. Uma PCR foi otimizada utilizando-se primers descritos em literatura para amplificar a região do gene SOD1. A amostra foi, então, submetida a sequenciamento unidirecional, que revelou que o animal em questão era homozigoto para o alelo A para a mutação c.118G>A no éxon 2 do gene SOD1. O diagnóstico clínico presuntivo da MDC no presente caso foi esclarecido por meio dos achados histopatológicos, associados aos achados clínicos, e da sua caracterização molecular. Ressalta-se a contribuição deste relato, que traz aspectos clínicos, histopatológicos e moleculares associados à MDC na raça Pastor-Suíço, para a qual, até o presente momento, na literatura consultada, não há relato dessa enfermidade.(AU)

The objective of this study was to describe the clinical, histopathological and molecular findings associated with MDC in a Swiss Shepherd dog. The dog had a progressive paraparesis in pelvic limbs and was submitted to clinical evaluations where, among other differentials, the presumptive diagnosis of MDC was obtained. With the progression of the nervous deficits tutor opted for euthanasia. The histopathological findings of the spinal cord were compatible with axonal and myelinic segmental degeneration. Molecular diagnosis was performed by extracting the DNA obtained by oral swab. PCR was optimized using primers described in the literature to amplify the SOD1 gene region. The sample was then subjected to one-way sequencing which revealed that the animal in question was homozygous for the A allele for the c.118G>A mutation in exon 2 of the SOD1 gene. The presumptive diagnosis of MDC in the present case was clarified by histopathological findings, as well as by its molecular characterization. The contribution of this report brings clinical, histopathological and molecular aspects associated with canine degenerative myelopathy in the Swiss Shepherd breed, that until this moment, in the literature consulted, there is no report of this disease in the breed mentioned.(AU)

Adjuvant-induced arthritis affects testes and ventral prostate of Wistar rats.

BACKGROUND: Rheumatoid arthritis (RA) may reduce the testosterone production, thereby leading to testicular dysfunction and subfertility. OBJECTIVES: This study aimed to evaluate whether adjuvant-induced arthritis (AIA) induces histopathological and morphometric-stereological alterations on testes with repercussions on the prostate, and alternatively, verifying AIA-induced direct effects on the prostate, regardless of the testicular involvement. MATERIAL AND METHODS: Adult male Wistar rats were sham-orchiectomized or orchiectomized. Twenty days after the surgery, these animals were injected with vehicle (SHAM and ORQ groups, respectively) or adjuvant (Mycobacterium tuberculosis) to induce arthritis (AIA and ORQ/AIA groups, respectively). Forty days later, testes and ventral prostate were processed for histopathological and morphometric-stereological analyses, as well as to PCNA immunohistochemistry. Collagen deposit was evaluated in prostate. Circulating testosterone levels were determined 15 days post-AIA induction in SHAM and AIA rats and 40th day in all groups. RESULTS: In the testes, AIA promoted histopathological changes characterized by an increase in the percentage of abnormal tubules and reduction in the height of the seminiferous epithelium, daily production of spermatozoa, and cellular proliferation. In the prostate, AIA decreased the luminal volume of the secretory ducts. In condition of androgenic deprivation due to the orchiectomy, AIA induced proliferation of the prostatic epithelium. DISCUSSION: The effects of arthritis on testes and prostate were observed 40 days post-AIA induction, possibly results of the hypoandrogenism were already established on 15th day post-induction, which is related to the decline of the steroidogenesis in the Leydig cells. On the other hand, the joint inflammatory process may also have direct repercussions upon the prostate, regardless of this hypoandrogenism. CONCLUSION: AIA effects on reproductive tissues may be related to both hypoandrogenism and other direct inflammatory mechanisms. Possibly, these AIA effects on the testes and prostate occur at a stage in which the inflammatory process is most active, about 15-20 days after induction, remaining evident until the 40th day.

The senses of the choroid plexus.

The composition of cerebrospinal and brain interstitial fluids is ensured by barriers between the blood and the brain parenchyma (the blood-brain barrier) and between the blood and the cerebrospinal fluid (the blood-cerebrospinal fluid barrier). Barrier function results from the combination of tight junctions between cells that impair solute flux via the paracellular pathway, cell membrane transporters that enable selective transcellular solute passage, and intracellular metabolizing enzymes that transform molecules in transit. Collectively, they comprise a chemical surveillance system, essential to protect the brain from toxicants, microorganisms, and other harmful compounds. Conversely, this chemical surveillance system compromises the brain delivery of many pharmacologic agents against brain cancer and brain metastasis, neurodegenerative diseases, and brain infections. Despite their importance, the mechanisms underlying the regulation of the components of this chemical surveillance system in response to alterations in the composition of blood and brain fluids are still poorly understood. We propose that odorant receptors, vomeronasal receptors and taste receptors, recently identified at brain barriers might be upstream components of this surveillance system. These chemosensory receptors are strategically placed to monitor the composition of blood, cerebrospinal and brain interstitial fluids. Upon ligand-binding, they may deploy the action of transporters and detoxifying enzymes or other unprecedented functions in brain barrier cells, to cope with alterations in the composition of blood and brain cerebrospinal and interstitial fluids, working as guardians of the central nervous system.

Treating TNF Receptor Associated Periodic Fever Syndrome in End-Stage Renal Failure.

Tumor necrosis factor receptor associated periodic syndrome (TRAPS) is a rare monogenic autoinflammatory disease. Its most severe manifestation is secondary amyloidosis. A 44-year-old male presented with nephrotic syndrome. Kidney biopsy was conclusive for secondary amyloidosis. The patient and his children had a history of recurrent febrile periods since infancy. All subjects were positive for a heterozygous variant of the TNFRSF1A gene, confirming TRAPS diagnosis. The patient progressed to end-stage renal failure and developed recurrent pericarditis episodes. He was started on anakinra while on hemodialysis with marked reduction of his serum amyloid A protein (SAA) levels. Meanwhile he received a cadaveric renal transplant and maintains anakinra treatment. Despite renal failure being the most feared complication of AA amyloidosis caused by TRAPS, little data is available about safety of anti-IL-1 treatment in patients with severe kidney failure. The authors report this case of a patient on dialysis treated with anakinra in which no complications were registered. Though amyloidosis is established, the authors believe containing its progression and reducing inflammatory activity can improve patient prognosis and reduce recurrence of amyloidosis in kidney transplant, as has been demonstrated in transplanted patients due to familial Mediterranean fever amyloidosis.

Acupuncture on anxiety and inflammatory events following surgery of mandibular third molars: a split-mouth, randomized, triple-blind clinical trial.

The aim of this study was to compare the effects of acupuncture and placebo acupuncture on the control of pain, oedema, and trismus following the extraction of third molars and on the control of preoperative anxiety. Sixteen patients (mean age 22.5±3.45years) each underwent four acupuncture sessions, one prior to each surgery and the others at 24, 48, and 72hours after each surgery (left and right tooth). Oedema was determined using measurements of the face and trismus was determined by maximum mouth opening at baseline and at 24, 48, 72hours and 7days following surgery. Postoperative pain was evaluated by the patients using a visual analogue scale (VAS) at 24, 48, and 72hours following surgery. Anxiety was evaluated using the State-Trait Anxiety Inventory and a VAS at baseline and before and after acupuncture prior to surgery. The statistical analysis was performed using the paired t-test and Wilcoxon test. Acupuncture showed a better performance in the control of oedema at 48hours (P=0.026), 72hours (P=0.046), and 7days (P=0.040) when compared to placebo. There was no statistically significant difference between the acupuncture and placebo groups in the control of pain, trismus, or anxiety.

Metal loads and biomarker suite responses in a tropical carnivorous fish indicative of anthropogenic impacts in a Southeastern Brazilian lagoon.

Tropical coastal lagoons are highly productive environments exhibiting high biodiversity. However, the use of these ecosystems by local communities is of concern, since this generally leads to environmental degradation. The Imboassica coastal lagoon, located in Macaé city, in Northern Rio de Janeiro, is an important ecosystem in the state, however, already displaying signs of anthropogenic impacts. Carnivorous fish Hoplias malabaricus specimens were sampled from this impacted site, as well as from a reference area. Fish from Imboassica Lagoon presented lower condition factor, lower cholinesterase activity, and higher percentage of erythrocyte micronuclei when compared to fish from the reference site. Metals in fish from Imboassica Lagoon were always higher than Encantada Lagoon, with some seasonal differences, where some metals were higher in the rainy season compared to the dry season in muscle tissue, with the exception of Cu, Fe, Sr, and Zn; and in the liver, except for Ba, Cd, Cr, Ni, and Sr. Cr and Mn in the edible muscle portion of the fish were higher than the limits established by Brazilian and International legislations as permissible for human consumption, thus leading to concerns regarding public health risks for the local population that use fish as their main protein source.

Performance evaluation of a natural treatment system for small communities, composed of a UASB reactor, maturation ponds (baffled and unbaffled) and a granular rock filter in series.

Post-treatment of anaerobic reactor effluent with maturation ponds is a good option for small to medium-sized communities in tropical climates. The treatment line investigated, operating in Brazil, with an equivalent capacity to treat domestic sewage from 250 inhabitants, comprised a upflow anaerobic sludge blanket reactor followed by two shallow maturation ponds (unbaffled and baffled) and a granular rock filter (decreasing grain size) in series, requiring an area of only 1.5 m2 inhabitant-1. With an overall hydraulic retention time of only 6.7 days, the performance was excellent for a natural treatment system. Based on over two years of continuous monitoring, median removal efficiencies were: biochemical oxygen demand = 93%, chemical oxygen demand = 79%, total suspended solids = 87%, ammonia = 43% and Escherichia coli = 6.1 log units. The final effluent complied with European discharge standards and WHO guidelines for some forms of irrigation, and appeared to be a suitable alternative for treating domestic sewage for small communities in warm areas, especially in developing countries.

Consequences of intraoperative spinal cord manipulation in dogs with thoracolumbar intervertebral disc extrusion / Consequências da manipulação medular intraoperatória em cães com extrusão intervertebral de disco toracolombar

The objective of the present study was to evaluate if extradural contact during hemilaminectomy would cause neurological deterioration in the early and/or late postoperative period in dogs with intervertebral disc extrusion. Nineteen dogs with thoracolumbar intervertebral disc extrusion underwent hemilaminectomy for spinal cord decompression and removal of extruded disc material. Meningeal contacts during surgery were quantified. Paraplegia (with nociception) and paraparesis were observed in 11/19 and 8/19 of dogs, respectively, before surgery. At the end of our study, only two (2/19) had paraplegia and one (1/19), paraparesis. There were more extradural contacts when extruded intervertebral disc material was at a ventrolateral position. Extradural contacts during surgery had no influence on neurological progression nor on time to recovery of motor function. Immediately (24 and 48 hours) after surgery, 13/19 dogs had the same neurological stage before surgery. At 7 and 90 days, 13/19 and 17/19 dogs, respectively, showed neurological improvement, compared with their preoperative stage. There was no influence of the number of extradural contacts on neurological recovery. These findings indicate that a careful inspection of the vertebral canal for removal of as much extruded disc material as possible does not cause neurologic deterioration.(AU)

O presente trabalho teve como objetivo avaliar se contatos extradurais durante hemilaminectomia em cães com extrusão de disco intervertebral causariam piora neurológica no pós-operatório imadiato e/ou tardio. Dezenove cães com extrusão toracolombar de disco intervertebral foram submetidos à hemilaminectomia para descompressão medular e remoção do material extruso. Durante o procedimento cirúrgico, os contatos meningomedulares foram quantificados. Antes da cirurgia, 11/19 cães apresentavam paraplegia (com nocicepção) e 8/19 cães, paraparesia. Ao fim do estudo, apenas dois cães (2/19) mostravam paraplegia com dor profunda e um (1/19), paraparesia. Observou-se maior quantidade de contatos extradurais quando o material discal extruso encontrava-se em posição ventrolateral. Os contatos extradurais não mostraram influência estatística na evolução neurológica dos animais, bem como no tempo de recuperação das funções motora. Vinte e quatro e 48 horas após a cirurgia, 13/19 cães apresentavam o mesmo grau neurológico de antes da cirurgia. Após sete e 90 dias de pós-operatório, 13/19 e 17/19 demonstraram melhora neurológica em comparação com o pré-operatório, respectivamente. A quantidade de contatos extradurais não influenciou na recuperação neurológica dos cães. Esses achados indicam que uma inspeção minuciosa do canal vertebral pode ser recomendada, a fim de remover o máximo de material discal extruso, evitando-se piora neurológica por compressão medular.(AU)

Aparência tomográfica dos discos intervertebrais mineralizados em cães da raça Dachshund Miniatura / Tomographic appearance of calcified intervertebral discs in miniature dachshund

Objetivou-se com este trabalho descrever os aspectos de tomografia computadorizada de discos intervertebrais de cães da raça Dachshund com um ano de idade, assim como quantificar, qualificar e localizar a mineralização nos componentes do DIV e no espaço intervertebral da coluna vertebral. Dez cães dessa raça, com um ano de idade, foram submetidos à tomografia computadorizada (TC). Nas imagens de reconstrução sagital dos animais, procedeu-se à identificação de sete DIV em cada animal, entre as vértebras T9 e L3, totalizando a análise de 70 DIV. Entre os DIV mineralizados, a presença da alteração foi determinada quanto à localização no núcleo pulposo (NP), no anel fibroso (AF) e em AF/NP. Dos 70 DIV avaliados, 45 foram considerados como mineralizados. Entre os classificados como mineralizados, 20% (9/45), 17,8% (8/45) e 62,2% (28/45) estavam localizados nas regiões do NP, AF e AF/NP, respectivamente. A mineralização encontrada neste estudo esteve caracterizada por aumento de atenuação radiográfica com densidade mineral na topografia do DIV, havendo variações da localização e da radiodensidade de área calcificada entre animais e DIV no mesmo indivíduo. As alterações relativas à mineralização dos DIV podem ser classificadas tomograficamente quanto à localização da alteração no disco em AF, NP e AF/NP.(AU)

The objective was to describe CT imaging of calcified IVD in one-year-old Dachshunds, as well as describe the location in the soft tissue structures that make up the IVD. Ten one-year-old dogs underwent computed tomography (CT). In sagittal reconstruction images of animals, the identification of 70 IVD present in the spaces between L3 - T9 were performed. Among calcified IVDs, the presence of the change was determined as the location in the nucleus pulposus (NP), annulus fibrosus (AF) and both. Of a total of 70 evaluated IVDs, 45 were found to calcified and 25 not calcified. Among calcified discs, location was determined in nine (20%) in NP, eight discs (17.77%) in AF and 28 (62, 23%) NP & AF. Calcification in this study was characterized by increased radiographic attenuation mineral density in the IVD topography. There is variation in the location and radiodensity of calcified areas between animals and IVD in the same individual. Changes related to IVD calcification can be classified as tomographic location in AF, NP and NP & AF.(AU)

Influence of third molars in mandibular fractures. Part 2: mandibular condyle-a meta-analysis.

The aim of this systematic review was to investigate the influence of the presence and position of mandibular third molars in mandibular condyle fractures. An electronic search was conducted in PubMed, Scopus, Web of Science, Cochrane Library, and VHL, through January 2016. The eligibility criteria included observational studies. The search strategy resulted in 704 articles. Following the selection process, 13 studies were included in the systematic review and 11 in the meta-analysis. In terms of the risk of bias analysis, six studies presented ≤6 stars in the Newcastle-Ottawa scale assessment. The presence of a mandibular third molar decreased the probability of condylar fracture (cross-sectional and case-control studies: odds ratio (OR) 0.26, 95% confidence interval (CI) 0.17-0.40, I2=87.8%; case-control studies: OR 0.30, 95% CI 0.16-0.58, I2=91.6%). The third molar positions most favourable to condylar fracture according to the Pell and Gregory classification are class A (OR 1.32, 95% CI 1.09-1.61, I2=0%) and class I (OR 1.37, 95% CI 1.05-1.77, I2=32.8%). Class B (OR 0.69, 95% CI 0.49-0.97, I2=56.0%) and class II (OR 0.71, 95% CI 0.57-0.87, I2=0%) act as protective factors for condylar fracture. The results suggest that the presence of a mandibular third molar decreases the chance of condylar fracture and that the positions of the third molar most favourable for condylar fracture are classes A and I, with classes B and II acting as protective factors.

Influence of third molars in mandibular fractures. Part 1: mandibular angle-a meta-analysis.

The aim of this systematic review was to investigate the influence of the presence and position of mandibular third molars on angle fractures. An electronic search was conducted in the PubMed, Scopus, Web of Science, Cochrane Library, and VHL databases, through January 2016. The eligibility criteria included observational studies. The search strategy resulted in 704 articles. Following the selection process, 35 studies were included in the systematic review and 28 in the meta-analysis. Twenty studies presented a score of ≤6 stars in the Newcastle-Ottawa scale assessment, indicating a risk of bias in the analysis. The presence of a mandibular third molar increases the chance of an angle fracture (case-control and cross-sectional studies: odds ratio (OR) 3.83, 95% confidence interval (CI) 3.02-4.85, I2=83.1%; case-control studies: OR 3.27, 95% CI 2.57-4.16, I2=81.3%). The third molar positions most favourable to angle fracture according to the Pell and Gregory classification are class B (OR 1.44, 95% CI 1.06-1.96, I2=87.2%) and class II (OR 1.67, 95% CI 1.36-2.04, I2=72.4%). Class A (OR 0.60, 95% CI 0.45-0.81, I2=87.1%) and class I (OR 0.51, 95% CI 0.37-0.71, I2=89.4%) act as protective factors for angle fracture. The results suggest that the presence of the third molar increases the chance of angle fracture by 3.27 times and that the most favourable positions of the third molar for angle fracture are classes B and II, whilst classes A and I act as protective factors.

Evaluation of a Brain Acetylcholinesterase Extraction Method and Kinetic Constants after Methyl-Paraoxon Inhibition in Three Brazilian Fish Species.

Acetylcholinesterase (AChE) is an important enzyme in the control of the neuronal action potential and sensitive to organophosphate inhibition. Brain fish AChE is less sensitive to organophosphate inhibition than AChE from terrestrial animals, although this sensitivity is variable among species and has not yet been fully evaluated in fish species. In this setting, inhibition kinetic constants for progressive irreversible inhibition of brain acetylcholinesterase due to methyl-paraoxon exposure were determined in three fish species (Mugil liza, Genidens genidens and Lagocephalus laevigatus) and hen (Gallus domesticus). Enzyme extraction using a detergent was shown to be adequate, and samples presented activity inhibition in high substrate concentrations and suppression of inhibition by methyl-paraoxon in the presence of the substrate, similar to kinetic patterns from purified enzyme preparations. Catfish (G. genidens) AChE presented the highest sensitivity among the evaluated fish species (IC50 = 1031.20 nM ± 63.17) in comparison to M. liza and L. laevigatus (IC50: 2878.83 ± 421.94 and 2842.5 ± 144.63 nM respectively). The lower dissociation constant (Kd = 20.3 ± 2.95 µM) of catfish AChE showed greater enzyme affinity for methyl-paraoxon, explaining this species higher sensitivity to organophosphates. Hen AChE presented higher ki (900.57 ± 65.3 mM-1min-1) and, consequently, greater sensitivity to methyl-paraoxon, explained by a lower Kd (0.6 ± 0.13 µM). Furthermore, hen AChE did not differentiate between the propionylthiocholine and acetylthiocholine substrates, indicating easier access of methyl-paraoxon to the hen enzyme activity site. The results obtained herein indicate a suitable extraction of AChE and, despite different inhibition kinetic constants, demonstrate that fish AChE is less sensitive to methyl-paraoxon, probably due to reduced access to the catalytic center which provides greater enzyme substrate selectivity.

Sex Hormone Decline and Amyloid ß Synthesis, Transport and Clearance in the Brain.

Sex hormones (SH) are essential regulators of the central nervous system. The decline in SH levels along with ageing may contribute to compromised neuroprotection and set the grounds for neurodegeneration and cognitive impairments. In Alzheimer's disease, besides other pathological features, there is an imbalance between amyloid ß (Aß) production and clearance, leading to its accumulation in the brain of older subjects. Aß accumulation is a primary cause for brain inflammation and degeneration, as well as concomitant cognitive decline. There is mounting evidence that SH modulate Aß production, transport and clearance. Importantly, SH regulate most of the molecules involved in the amyloidogenic pathway, their transport across brain barriers for elimination, and their degradation in the brain interstitial fluid. This review brings together data on the regulation of Aß production, metabolism, degradation and clearance by SH.

Sex Hormones Protect Against Amyloid-ß Induced Oxidative Stress in the Choroid Plexus Cell Line Z310.

The choroid plexus (CP) epithelium is a unique structure in the brain that forms an interface between the peripheral blood on the basal side and the cerebrospinal fluid (CSF) on the apical side. It is a relevant source of many polypeptides secreted to the CSF with neuroprotective functions and also participates in the elimination and detoxification of brain metabolites, such as ß-amyloid (Aß) removal from the CSF through transporter-mediated influx. The CP is also a target tissue for sex hormones (SHs) that have recognised neuroprotective effects against a variety of insults, including Aß toxicity and oxidative stress in the central nervous system. The present study aimed to understand how SHs modulate Aß-induced oxidative stress in a CP cell line (Z310 cell line) by analysing the effects of Aß1-42 on oxidative stress, mitochondrial function and apoptosis, as well as by assessing how 17ß-oestradiol (E2 ) and 5α-dihydrotestosterone (DHT) modulated these effects and the cellular uptake of Aß1-42 by CP cells. Our findings show that E2 and DHT treatment reduce Aß1-42 -induced oxidative stress and the internalisation of Aß1-42 by CP epithelial cells, highlighting the importance of considering the background of SHs and therefore sex-related differences in Aß metabolism and clearance by CP cells.

The dark and bright sides of an enzyme: a three dimensional structure of the N-terminal domain of Zophobas morio luciferase-like enzyme, inferences on the biological function and origin of oxygenase/luciferase activity.

Beetle luciferases, the enzymes responsible for bioluminescence, are special cases of CoA-ligases which have acquired a novel oxygenase activity, offering elegant models to investigate the structural origin of novel catalytic functions in enzymes. What the original function of their ancestors was, and how the new oxygenase function emerged leading to bioluminescence remains unclear. To address these questions, we solved the crystal structure of a recently cloned Malpighian luciferase-like enzyme of unknown function from Zophobas morio mealworms, which displays weak luminescence with ATP and the xenobiotic firefly d-luciferin. The three dimensional structure of the N-terminal domain showed the expected general fold of CoA-ligases, with a unique carboxylic substrate binding pocket, permitting the binding and CoA-thioesterification activity with a broad range of carboxylic substrates, including short-, medium-chain and aromatic acids, indicating a generalist function consistent with a xenobiotic-ligase. The thioesterification activity with l-luciferin, but not with the d-enantiomer, confirms that the oxygenase activity emerged from a stereoselective impediment of the thioesterification reaction with the latter, favoring the alternative chemiluminescence oxidative reaction. The structure and site-directed mutagenesis support the involvement of the main-chain amide carbonyl of the invariant glycine G323 as the catalytic base for luciferin C4 proton abstraction during the oxygenase activity in this enzyme and in beetle luciferases (G343).

"Tasting" the cerebrospinal fluid: Another function of the choroid plexus?

The choroid plexus (CP) located in brain ventricles, by forming the interface between the blood and the cerebrospinal fluid (CSF) is in a privileged position to monitor the composition of these body fluids. Yet, the mechanisms involved in this surveillance system remain to be identified. The taste transduction pathway senses some types of molecules, thereby evaluating the chemical content of fluids, not only in the oral cavity but also in other tissues throughout the body, such as some cell types of the airways, the gastrointestinal tract, testis and skin. Therefore, we hypothesized that the taste transduction pathway could also be operating in the CP to assess the composition of the CSF. We found transcripts for some taste receptors (Tas1r1, Tas1r2, Tas1r3, Tas2r109 and Tas2r144) and for downstream signaling molecules (α-Gustducin, Plcß2, ItpR3 and TrpM5) that encode this pathway, and confirmed the expression of the corresponding proteins in Wistar rat CP explants and in the CP epithelial cells (CPEC). The functionality of the T2R receptor expressed in CP cells was assessed by calcium imaging, of CPEC stimulated with the bitter compound D-Salicin, which elicited a rise in the intracellular Ca(2+). This effect was diminished in the presence of the bitter receptor blocker Probenecid. In summary, we described the expression of the taste-related components involved in the transduction signaling cascade in CP. Taken together, our results suggest that the taste transduction pathway in CPEC makes use of T2R receptors in the chemical surveillance of the CSF composition, in particular to sense bitter noxious compounds.

Sex-Related Differences in Rat Choroid Plexus and Cerebrospinal Fluid: A cDNA Microarray and Proteomic Analysis.

The choroid plexus (CP) epithelium is a unique structure in the brain that forms an interface between the peripheral blood and the cerebrospinal fluid (CSF), which is mostly produced by the CP itself. Because the CP transcriptome is regulated by the sex hormone background, the present study compared gene/protein expression profiles in the CP and CSF from male and female rats aiming to better understand sex-related differences in CP functions and brain physiology. We used data previously obtained by cDNA microarrays to compare the CP transcriptome between male and female rats, and complemented these data with the proteomic analysis of the CSF of castrated and sham-operated males and females. Microarray analysis showed that 17 128 and 17 002 genes are expressed in the male and female CP, which allowed the functional annotation of 141 and 134 pathways, respectively. Among the most expressed genes, canonical pathways associated with mitochondrial dysfunctions and oxidative phosphorylation were the most prominent, whereas the most relevant molecular and cellular functions annotated were protein synthesis, cellular growth and proliferation, cell death and survival, molecular transport, and protein trafficking. No significant differences were found between males and females regarding these pathways. Seminal functions of the CP differentially regulated between sexes were circadian rhythm signalling, as well as several canonical pathways related to stem cell differentiation, metabolism and the barrier function of the CP. The proteomic analysis identified five down-regulated proteins in the CSF samples from male rats compared to females and seven proteins exhibiting marked variation in the CSF of gonadectomised males compared to sham animals, whereas no differences were found between sham and ovariectomised females. These data clearly show sex-related differences in CP gene expression and CSF protein composition that may impact upon neurological diseases.

Tumour-suppression function of KLF12 through regulation of anoikis.

Suppression of detachment-induced cell death, known as anoikis, is an essential step for cancer metastasis to occur. We report here that expression of KLF12, a member of the Kruppel-like family of transcription factors, is downregulated in lung cancer cell lines that have been selected to grow in the absence of cell adhesion. Knockdown of KLF12 in parental cells results in decreased apoptosis following cell detachment from matrix. KLF12 regulates anoikis by promoting the cell cycle transition through S phase and therefore cell proliferation. Reduced expression levels of KLF12 results in increased ability of lung cancer cells to form tumours in vivo and is associated with poorer survival in lung cancer patients. We therefore identify KLF12 as a novel metastasis-suppressor gene whose loss of function is associated with anoikis resistance through control of the cell cycle.