Fabrication and characterization of a bioactive polymethylmethacrylate-based porous cement loaded with strontium/calcium apatite nanoparticles.

Polymethylmethacrylate (PMMA)-based cements are used for bone reparation due to their biocompatibility, suitable mechanical properties, and mouldability. However, these materials suffer from high exothermic polymerization and poor bioactivity, which can cause the formation of fibrous tissue around the implant and aseptic loosening. Herein, we tackled these problems by adding Sr2+ -substituted hydroxyapatite nanoparticles (NPs) and a porogenic compound to the formulations, thus creating a microenvironment suitable for the proliferation of osteoblasts. The NPs resembled the structure of the bone's apatite and enabled the controlled release of Sr2+ . Trends in the X-ray patterns and infrared spectra confirmed that Sr2+ replaced Ca2+ in the whole composition range of the NPs. The inclusion of an effervescent additive reduced the polymerization temperature and lead to the formation of highly porous cement exhibiting mechanical properties comparable to the trabecular bone. The formation of an opened and interconnected matrix allowed osteoblasts to penetrate the cement structure. Most importantly, the gas formation confined the NPs at the surface of the pores, guaranteeing the controlled delivery of Sr2+ within a concentration sufficient to maintain osteoblast viability. Additionally, the cement was able to form apatite when immersed into simulated body fluids, further increasing its bioactivity. Therefore, we offer a formulation of PMMA cement with improved in vitro performance supported by enhanced bioactivity, increased osteoblast viability and deposition of mineralized matrix assigned to the loading with Sr2+ -substituted hydroxyapatite NPs and the creation of an interconnected porous structure. Altogether, our results hold promise for enhanced bone reparation guided by PMMA cements.

A new approach for the determination of 210Pb by liquid scintillation counting.

A new approach to the calibration procedure is presented, as there is no need to discount bismuth from lead spectrum when constructing the efficiency curve. This work presents two calibration methods: one considering mostly lead spectrum contributions and other that considers both lead and bismuth contributions. Both methodologies provide consistent results when evaluated in an intercomparison program. Furthermore, this methodology allows simultaneous analysis of several samples and is suitable for any type of sample after proper digestion in liquid form.

Lipocalin-2 regulates adult neurogenesis and contextual discriminative behaviours.

In the adult mammalian brain, newborn granule cells are continuously integrated into hippocampal circuits, and the fine-tuning of this process is important for hippocampal function. Thus, the identification of factors that control adult neural stem cells (NSCs) maintenance, differentiation and integration is essential. Here we show that the deletion of the iron trafficking protein lipocalin-2 (LCN2) induces deficits in NSCs proliferation and commitment, with impact on the hippocampal-dependent contextual fear discriminative task. Mice deficient in LCN2 present an increase in the NSCs population, as a consequence of a G0/G1 cell cycle arrest induced by increased endogenous oxidative stress. Of notice, supplementation with the iron-chelating agent deferoxamine rescues NSCs oxidative stress, promotes cell cycle progression and improves contextual fear conditioning. LCN2 is, therefore, a novel key modulator of neurogenesis that, through iron, controls NSCs cell cycle progression and death, self-renewal, proliferation and differentiation and, ultimately, hippocampal function.

Lipocalin 2 modulates the cellular response to amyloid beta.

The production, accumulation and aggregation of amyloid beta (Aß) peptides in Alzheimer's disease (AD) are influenced by different modulators. Among these are iron and iron-related proteins, given their ability to modulate the expression of the amyloid precursor protein and to drive Aß aggregation. Herein, we describe that lipocalin 2 (LCN2), a mammalian acute-phase protein involved in iron homeostasis, is highly produced in response to Aß1-42 by choroid plexus epithelial cells and astrocytes, but not by microglia or neurons. Although Aß1-42 stimulation decreases the dehydrogenase activity and survival of wild-type astrocytes, astrocytes lacking the expression of Lcn2 are not affected. This protection results from a lower expression of the proapoptotic gene Bim and a decreased inflammatory response. Altogether, these findings show that Aß toxicity to astrocytes requires LCN2, which represents a novel mechanism to target when addressing AD.

The response of social anxiety disorder patients to threat scenarios differs from that of healthy controls

The objective of the present study was to evaluate the response of social anxiety disorder (SAD) patients to threat scenarios. First-choice responses to 12 scenarios describing conspecific threatening situations and mean scores of defensive direction and defensive intensity dimensions were compared between 87 SAD patients free of medication and 87 matched healthy controls (HC). A significant gender difference in the first-choice responses was identified for seven scenarios among HCs but only for two scenarios among SAD patients. A significantly higher proportion of SAD patients chose "freezing" in response to "Bush" and "Noise" scenarios, whereas the most frequent response by HCs to these scenarios was "check out". SAD males chose "run away" and "yell" more often than healthy men in response to the scenarios "Park" and "Elevator", respectively. There was a positive correlation between the severity of symptoms and both defensive direction and defensive intensity dimensions. Factorial analysis confirmed the gradient of defensive reactions derived from animal studies. SAD patients chose more urgent defensive responses to threat scenarios, seeming to perceive them as more dangerous than HCs and tending to move away from the source of threat. This is consistent with the hypothesis that the physiopathology of anxiety disorders involves brain structures responsible for defensive behaviors.

The response of social anxiety disorder patients to threat scenarios differs from that of healthy controls.

The objective of the present study was to evaluate the response of social anxiety disorder (SAD) patients to threat scenarios. First-choice responses to 12 scenarios describing conspecific threatening situations and mean scores of defensive direction and defensive intensity dimensions were compared between 87 SAD patients free of medication and 87 matched healthy controls (HC). A significant gender difference in the first-choice responses was identified for seven scenarios among HCs but only for two scenarios among SAD patients. A significantly higher proportion of SAD patients chose "freezing" in response to "Bush" and "Noise" scenarios, whereas the most frequent response by HCs to these scenarios was "check out". SAD males chose "run away" and "yell" more often than healthy men in response to the scenarios "Park" and "Elevator", respectively. There was a positive correlation between the severity of symptoms and both defensive direction and defensive intensity dimensions. Factorial analysis confirmed the gradient of defensive reactions derived from animal studies. SAD patients chose more urgent defensive responses to threat scenarios, seeming to perceive them as more dangerous than HCs and tending to move away from the source of threat. This is consistent with the hypothesis that the physiopathology of anxiety disorders involves brain structures responsible for defensive behaviors.

Ciclopirox olamine: an antifungal alternative against cryptococcosis.

AIMS: The in vitro activity of ciclopirox olamine was evaluated against Cryptococcus spp. obtained from the cerebrospinal fluid (CSF) of immunocompromised patients. METHODS AND RESULTS: The antifungal activity of ciclopirox olamine was tested against Cryptococcus spp. obtained from the CSF of immunocompromised patients, using amphotericin B and fluconazole as controls. The minimal inhibitory concentration was determined following the microdilution method indicated by the Clinical and Laboratory Standards Institute. The minimal fungicide concentration was determined by the absence of growth on Sabouraud dextrose agar. The data obtained showed that antifungal activity of ciclopirox olamine ranged from 0·25 to 1 µg ml(-1) . CONCLUSIONS: This paper underscores the importance of the antifungal potential of ciclopirox olamine against Cryptococcus spp. as an alternative treatment against systemic cryptococosis. In vivo experiments are essential for future medical use. SIGNIFICANCE AND IMPACT OF THE STUDY: This was the first time that ciclopirox olamine was tested against Cryptococcus spp. using the reference method. The antifungal activity of this drug against this species suggests an applicable potential for systemic cryptococcosis therapy.

A mobile bioassay laboratory for the assessment of internal doses based on in vivo and in vitro measurements.

Internal exposures may occur in nuclear power plants, radioisotope production, and in medicine and research laboratories. Such practices require quick response in case of accidents of a wide range of magnitudes. This work presents the design and calibration of a mobile laboratory for the assessment of accidents involving workers and the population as well as for routine monitoring. The system was set up in a truck with internal dimensions of 3.30 m × 1.60 m × 1.70 m and can identify photon emitters in the energy range of 100-3,000 keV in the whole body, organs, and in urine. A thyroid monitor consisting of a lead-collimated NaI(Tl)3" × 3" (7.62 × 7.62 cm) detector was calibrated with a neck-thyroid phantom developed at the IRD (Instituto de Radioproteção e Dosimetria). Whole body measurements were performed with a NaI(Tl)8" × 4" (20.32 × 10.16 cm) detector calibrated with a plastic-bottle phantom. Urine samples were measured with another NaI(Tl) 3" × 3" (7.62 × 7.62 cm) detector set up in a steel support. Standard solutions were provided by the National Laboratory for Metrology of Ionizing Radiation of the IRD. Urine measurements are based on a calibration of efficiency vs. energy for standard volumes. Detection limits were converted to minimum committed effective doses for the radionuclides of interest using standard biokinetic and dosimetric models in order to evaluate the applicability and limitations of the system. Sensitivities for high-energy activation and fission products show that the system is suitable for use in emergency and routine monitoring of individuals under risk of internal exposure by such radionuclides.

Methodology for radionuclides quantification through "in vitro" bioassay.

In Brazil, the radionuclides used for therapy are: (131)I, (153)Sm, (90)Y and (177)Lu, both for routine or research protocols. The radionuclide activity excreted by patients may be quantified by bioassay analysis and constitutes a powerful tool for individual treatment planning. The Bioassay Laboratory (LBIOVT) of the Institute of Radiation Protection and Dosimetry (IRD) has equipments for gamma and beta spectroscopy. These systems are calibrated in energy and efficiency using reference sources supplied by the National Laboratory of Radiation Metrology (LMNRI/IRD). The LBIOVT has operational procedures according ISO-ABNT-17025 recommendations and participates of international and national intercomparisons. The patient samples are collected immediately after radiopharmaceutical administrations, at the hospital or at the patient residence, and are handled, stored and transported according national radiation protection regulations. The radionuclide specific activity (Bq/L) is referenced to date and time of excretion, for the estimation of the individual biological half-live. The volume of excreta may carefully manipulated in order to avoid losses and misinterpretation in the activity quantification. The process of the LBIOVT accreditation and its participation in intercomparisons may guarantee the confidence of the results, allowing the minimization of the uncertainties in the individual monitoring.

Frequency of 22q11.2 microdeletion in sporadic non-syndromic tetralogy of Fallot cases.

BACKGROUND: Tetralogy of Fallot (TOF) is a congenital conotruncal heart defect commonly found in DiGeorge (DGS) and velocardiofacial (VCFS) syndromes. The deletion of chromosome 22q11 has also been demonstrated in sporadic or familial cases of TOF. The aim of the present study was to investigate the frequency of del22q11 in patients with non-syndromic TOF seen at a tertiary Pediatric Cardiology care center. METHOD: One hundred and twenty three non-syndromic TOF patients were selected and evaluated by history, physical examination and review of medical records. Venous blood was drawn for genomic DNA extraction after informed consent 22q11 microdeletion diagnosis was conducted through a standardized SNP genotyping assay and consecutive homozygosity mapping. Phenotype-genotype correlations regarding cardiac anatomy were conducted. RESULTS: We evaluated 123 non-syndromic TOF patients for a 22q11 deletion. 105 (85.4%) patients presented pulmonary stenosis and 18 (14.6%) had pulmonary atresia. Eight patients (6.5%) were found to have a deletion. Of the deleted patients, three (37.5%) presented pulmonary atresia. We have verified a tendency towards a higher prevalence of pulmonary atresia when comparing TOF patients with and without 22q11 microdeletion. CONCLUSIONS: 22q11.2 deletion in non-syndromic TOF patients is present in approximately 6% of patients. We suggest a tendency towards a higher prevalence of pulmonary atresia in non-syndromic TOF patients with 22q11 microdeletion. Molecular genetic screening of non-syndromic TOF patient may be important for the correct care of these patients and a more specific genetic diagnostic and counseling.

PCR screening for 22q11.2 microdeletion: development of a new cost-effective diagnostic tool.

BACKGROUND: Del22q11.2 syndrome is the most frequent known chromosomal microdeletion syndrome. Previous studies suggest that a substantial number of patients with congenital heart disease have a 22q11 deletion. The molecular diagnosis of Del22q11.2 is usually made by fluorescence in situ hybridization, an expensive and not widely available technique. We developed an efficient and cost-effective PCR SNP assay designed for the screening of 22q11.2 deletion through consecutive homozygosity. METHODS: Through the screening of dbSNP we have selected SNP markers located in the 22q11.2 microdeleted region. Population heterozygosities were determined in 213 normal individuals. Designed assays consisted of PCR amplification followed by restriction enzyme digestion. Fragments generated were visualized on agarose gel and genotyped. RESULTS: Selected markers were: rs5748411, rs2238778, rs4819523 and rs4680. All selected markers were localized in the 22q11.2 deleted region. Allele and genotype frequencies of all selected markers were under Hardy-Weinberg equilibrium. Selected SNPs were not in linkage disequilibrium. Predicted assay specificity was estimated to be 92.86% in the Brazilian population. CONCLUSIONS: The use of consecutive homozygosity in this SNP-based diagnostic test may be used as a cost-effective tool in reference molecular genetics laboratories.

High specificity PCR screening for 22q11.2 microdeletion in three different ethnic groups

Congenital heart defects are the most common of all human birth defects. Numerous studies have shown that a deletion within chromosome 22q11 is associated with DiGeorge syndrome and certain forms of sporadic congenital cardiovascular disease. We have determined the value of a PCR assay using markers D22S941, D22S944 and D22S264 designed for the screening of 22q11.2 deletion through consecutive homozygosity in an ethnically admixed urban population. The study population comprised 149 unrelated men and women from three different ethnic groups (white, mulatto and black). Test specificity for the overall population was estimated at 98.3 percent. We found no significant difference when comparing heterozygosity indices and ethnicity (P value = 0.43 (D22S944), 0.22 (D22S264), and 0.58 (D22S941)). There was no significant difference regarding assay specificity between the three different ethnic groups studied. This assay could constitute a cost-effective way to screen a large number of patients at increased risk, since PCR techniques are easily available, are fast, can be automatized, and are significantly less expensive than fluorescence in situ hybridization

High specificity PCR screening for 22q11.2 microdeletion in three different ethnic groups.

Congenital heart defects are the most common of all human birth defects. Numerous studies have shown that a deletion within chromosome 22q11 is associated with DiGeorge syndrome and certain forms of sporadic congenital cardiovascular disease. We have determined the value of a PCR assay using markers D22S941, D22S944 and D22S264 designed for the screening of 22q11.2 deletion through consecutive homozygosity in an ethnically admixed urban population. The study population comprised 149 unrelated men and women from three different ethnic groups (white, mulatto and black). Test specificity for the overall population was estimated at 98.3%. We found no significant difference when comparing heterozygosity indices and ethnicity (P value = 0.43 (D22S944), 0.22 (D22S264), and 0.58 (D22S941)). There was no significant difference regarding assay specificity between the three different ethnic groups studied. This assay could constitute a cost-effective way to screen a large number of patients at increased risk, since PCR techniques are easily available, are fast, can be automatized, and are significantly less expensive than fluorescence in situ hybridization.

Different TBX5 interactions in heart and limb defined by Holt-Oram syndrome mutations.

To better understand the role of TBX5, a T-box containing transcription factor in forelimb and heart development, we have studied the clinical features of Holt-Oram syndrome caused by 10 different TBX5 mutations. Defects predicted to create null alleles caused substantial abnormalities both in limb and heart. In contrast, missense mutations produced distinct phenotypes: Gly80Arg caused significant cardiac malformations but only minor skeletal abnormalities; and Arg237Gln and Arg237Trp caused extensive upper limb malformations but less significant cardiac abnormalities. Amino acids altered by missense mutations were located on the three-dimensional structure of a related T-box transcription factor, Xbra, bound to DNA. Residue 80 is highly conserved within T-box sequences that interact with the major groove of target DNA; residue 237 is located in the T-box domain that selectively binds to the minor groove of DNA. These structural data, taken together with the predominant cardiac or skeletal phenotype produced by each missense mutation, suggest that organ-specific gene activation by TBX5 is predicated on biophysical interactions with different target DNA sequences.

Sleep and respiratory patterns in the Djungarian hamster.

Djungarian hamsters adapted to a 12/12-h L/D photoperiod at 28 +/- 1 degrees C Tamb, implanted with electroencephalogram and electromyogram electrodes and fitted with pneumomanometers were monitored for 4 h on 5 consecutive days to obtain baseline sleep stages and respiration in the nontorpid hamster. Fifty-seven percent of recording time was spent in the sleep state with rapid eye movement (REM) sleep occupying 11% of the total sleep time. An analysis of successive 30-min epochs revealed that REM percentage and REM period (REMP) frequency in the first 30 min were significantly (p less than 0.0001) lower compared to the last 150 min, while REMP duration remained unchanged throughout the recording session. Respiratory frequency was significantly higher (p less than 0.01) by 24% in REM sleep as compared to Non-REM sleep. The euthermic sleep pattern in the non-torpid Djungarian hamster, adapted to a steady-state photoperiod, contained alternating periods of non-rapid eye movement (Non-REM) and REM sleep, similar to that seen in other rodents. REM sleep pattern regulation became steady after the initial 90-min transition period during which REM frequency but not duration increased. This transition to a steady REM sleep pattern might allow flexibility in entering the torpor state during the presence of unfavorable environmental stimuli.

Microdialysis as an approach to quantitate the release of neuropeptides.

1. In vivo microdialysis was performed on anesthetized and awake rats to measured release of cholecystokinin from the posterior nucleus accumbens. 2. Basal levels of cholecystokinin were detectable by radioimmunoassay in some, but not all animals. 3. Recovery through the microdialysis probe ranged from 0.1-2.4% for cholecystokinin, suggesting practical limitations to this approach with present technology.

Lidocaine excretion in breast milk.

Nursing mothers are occasionally treated with intravenous lidocaine for ventricular dysrhythmias. There have been no reports on the excretion of lidocaine into breast milk. This case documents the excretion of lidocaine into breast milk in small amounts and shows the validity of the TDx methodology used in the whole-milk lidocaine assay. We observed breast-milk concentrations of lidocaine at 40 percent of the serum levels. Clinical practitioners should be aware the lidocaine is excreted into breast milk in small amounts and the mother could probably continue to safely breast-feed her child while on parenteral lidocaine. Any adverse reactions in the nursing infant would probably be limited to an idiosyncratic or allergic reaction.

Espondilartrose cervical. Aspectos neurologicos e incidencia em um decenio. / Cervical spondylosis.Neurologic aspects and incidence during a decenium

Sao revistos os apectos neurologicos da espondilartrose cervical. E estudada em seguida a incidencia dessa patologia no decenio 1971-80 no Hospital da Clinicas da Univesidade de Pernambuco, comparando-se os achados clinicos desse material com o estudado por Maia, na mesma regiao, na decada de 50