Optical constraints on two-photon voltage imaging.

Significance: Genetically encoded voltage indicators (GEVIs) are a valuable tool for studying neural circuits in vivo, but the relative merits and limitations of one-photon (1P) versus two-photon (2P) voltage imaging are not well characterized. Aim: We consider the optical and biophysical constraints particular to 1P and 2P voltage imaging and compare the imaging properties of commonly used GEVIs under 1P and 2P excitation. Approach: We measure the brightness and voltage sensitivity of voltage indicators from commonly used classes under 1P and 2P illumination. We also measure the decrease in fluorescence as a function of depth in the mouse brain. We develop a simple model of the number of measurable cells as a function of reporter properties, imaging parameters, and desired signal-to-noise ratio (SNR). We then discuss how the performance of voltage imaging would be affected by sensor improvements and by recently introduced advanced imaging modalities. Results: Compared with 1P excitation, 2P excitation requires ∼ 10 4 -fold more illumination power per cell to produce similar photon count rates. For voltage imaging with JEDI-2P in the mouse cortex with a target SNR of 10 (spike height to baseline shot noise), a measurement bandwidth of 1 kHz, a thermally limited laser power of 200 mW, and an imaging depth of > 300 µ m , 2P voltage imaging using an 80-MHz source can record from no more than ∼ 12 neurons simultaneously. Conclusions: Due to the stringent photon-count requirements of voltage imaging and the modest voltage sensitivity of existing reporters, 2P voltage imaging in vivo faces a stringent tradeoff between shot noise and tissue photodamage. 2P imaging of hundreds of neurons with high SNR at a depth of > 300 µ m will require either major improvements in 2P GEVIs or qualitatively new approaches to imaging.

Environmental stressor assessment of hydrocarbonoclastic bacteria biofilms from a marine oil spill.

The environmental and economic impact of an oil spill can be significant. Biotechnologies applied during a marine oil spill involve bioaugmentation with immobilised or encapsulated indigenous hydrocarbonoclastic species selected under laboratory conditions to improve degradation rates. The environmental factors that act as stressors and impact the effectiveness of hydrocarbon removal are one of the challenges associated with these applications. Understanding how native microbes react to environmental stresses is necessary for effective bioaugmentation. Herein, Micrococcus luteus and M. yunnanensis isolated from a marine oil spill mooring system showed hydrocarbonoclastic activity on Maya crude oil in a short time by means of total petroleum hydrocarbons (TPH) at 144 h: M. luteus up to 98.79 % and M. yunnanensis 97.77 % removal. The assessment of Micrococcus biofilms at different temperature (30 °C and 50 °C), pH (5, 6, 7, 8, 9), salinity (30, 50, 60, 70, 80 g/L), and crude oil concentration (1, 5, 15, 25, 35 %) showed different response to the stressors depending on the strain. According to response surface analysis, the main effect was temperature > salinity > hydrocarbon concentration. The hydrocarbonoclastic biofilm architecture was characterised using scanning electron microscopy (SEM) and atomic force microscopy (AFM). Subtle but significant differences were observed: pili in M. luteus by SEM and the topographical differences measured by AFM Power Spectral Density (PSD) analysis, roughness was higher in M. luteus than in M. yunnanensis. In all three domains of life, the Universal Stress Protein (Usp) is crucial for stress adaptation. Herein, the uspA gene expression was analysed in Micrococcus biofilm under environmental stressors. The uspA expression increased up to 2.5-fold in M. luteus biofilms at 30 °C, and 1.3-fold at 50 °C. The highest uspA expression was recorded in M. yunnanensis biofilms at 50 °C with 2.5 and 3-fold with salinities of 50, 60, and 80 g/L at hydrocarbon concentrations of 15, 25, and 35 %. M. yunnanensis biofilms showed greater resilience than M. luteus biofilms when exposed to harsh environmental stressors. M. yunnanensis biofilms were thicker than M. luteus biofilms. Both biofilm responses to environmental stressors through uspA gene expression were consistent with the behaviours observed in the response surface analyses. The uspA gene is a suitable biomarker for assessing environmental stressors of potential microorganisms for bioremediation of marine oil spills and for biosensing the ecophysiological status of native microbiota in a marine petroleum environment.

Thrombotic Event Prevention in Patients Undergoing Posterior Lumbar Arthrodesis: our experiencie. A retrospective case series study. / Prevención de eventos trombóticos en pacientes sometidos a cirugía de artrodesis lumbar por vía posterior: experiencia en nuestro centro. Estudio retrospectivo de casos.

INTRODUCTION: There is no generalized consensus regarding perioperative prophylaxis of venous thromboembolism (VTE), either on using or timing it in patients undergoing spine surgery. VTE is a current concern because, even though being an uncommon event, it can cause serious complications. The aim of the present study is to propose guidelines for the prevention of thrombotic events in posterior spinal surgery, either as deep vein thrombosis or pulmonary thromboembolism. If the number of patients getting prophylaxis drugs is reduced a subsequent reduction of the incidence of epidural hematoma can be expected. MATERIALS AND METHODS: A number of 235 patients who had undergone posterior spinal arthrodesis in the previous five years were studied. Mechanical thromboprophylaxis measures consisting of compression stockings were applied in all of them. Anticoagulant drugs were also applied whenever risk factors for thrombosis were observed. Early weight-bearing was resumed immediately after surgery. Demographic, clinical, and surgical variables were collected, as well as complications appearing during the follow-up period, that was scheduled at one, two, four, six and twelve months after the surgery. Thrombotic events, if present, were diagnosed by clinical and imaging tests such as ultrasound and CT angiography. RESULTS: From the total 235 patients of this series, one hundred and fifty-three cases met the study inclusion criteria. A total of four thrombotic events appeared, one in the form of deep vein thrombosis and other three in the form of pulmonary thromboembolism. These last patients suffering an embolism died because of it. None of the variables studied had statistical significance for the occurrence of a thrombotic event. All four patients who suffered thrombotic events were receiving anticoagulant drugs, in addition to mechanical compression stockings, because of the presence of risk factors for thrombosis. CONCLUSIONS: By applying the fore mentioned protocol, adequate prevention of thromboembolic events was achieved in this study population of patients undergoing posterior spinal surgery.

Unrecognized volatile and semi-volatile organic compounds from brake wear.

Motor vehicles are among the major sources of pollutants and greenhouse gases in urban areas and a transition to "zero emission vehicles" is underway worldwide. However, emissions associated with brake and tire wear will remain. We show here that previously unrecognized volatile and semi-volatile organic compounds, which have a similarity to biomass burning emissions are emitted during braking. These include greenhouse gases or, these classified as Hazardous Air Pollutants, as well as nitrogen-containing organics, nitrogen oxides and ammonia. The distribution and reactivity of these gaseous emissions are such that they can react in air to form ozone and other secondary pollutants with adverse health and climate consequences. Some of the compounds may prove to be unique markers of brake emissions. At higher temperatures, nucleation and growth of nanoparticles is also observed. Regions with high traffic, which are often disadvantaged communities, as well as commuters can be impacted by these emissions even after combustion-powered vehicles are phased out.

Nirmatrelvir for Vaccinated or Unvaccinated Adult Outpatients with Covid-19.

BACKGROUND: Nirmatrelvir in combination with ritonavir is an antiviral treatment for mild-to-moderate coronavirus disease 2019 (Covid-19). The efficacy of this treatment in patients who are at standard risk for severe Covid-19 or who are fully vaccinated and have at least one risk factor for severe Covid-19 has not been established. METHODS: In this phase 2-3 trial, we randomly assigned adults who had confirmed Covid-19 with symptom onset within the past 5 days in a 1:1 ratio to receive nirmatrelvir-ritonavir or placebo every 12 hours for 5 days. Patients who were fully vaccinated against Covid-19 and who had at least one risk factor for severe disease, as well as patients without such risk factors who had never been vaccinated against Covid-19 or had not been vaccinated within the previous year, were eligible for participation. Participants logged the presence and severity of prespecified Covid-19 signs and symptoms daily from day 1 through day 28. The primary end point was the time to sustained alleviation of all targeted Covid-19 signs and symptoms. Covid-19-related hospitalization and death from any cause were also assessed through day 28. RESULTS: Among the 1296 participants who underwent randomization and were included in the full analysis population, 1288 received at least one dose of nirmatrelvir-ritonavir (654 participants) or placebo (634 participants) and had at least one postbaseline visit. The median time to sustained alleviation of all targeted signs and symptoms of Covid-19 was 12 days in the nirmatrelvir-ritonavir group and 13 days in the placebo group (P = 0.60). Five participants (0.8%) in the nirmatrelvir-ritonavir group and 10 (1.6%) in the placebo group were hospitalized for Covid-19 or died from any cause (difference, -0.8 percentage points; 95% confidence interval, -2.0 to 0.4). The percentages of participants with adverse events were similar in the two groups (25.8% with nirmatrelvir-ritonavir and 24.1% with placebo). In the nirmatrelvir-ritonavir group, the most commonly reported treatment-related adverse events were dysgeusia (in 5.8% of the participants) and diarrhea (in 2.1%). CONCLUSIONS: The time to sustained alleviation of all signs and symptoms of Covid-19 did not differ significantly between participants who received nirmatrelvir-ritonavir and those who received placebo. (Supported by Pfizer; EPIC-SR ClinicalTrials.gov number, NCT05011513.).

Serum Troponin I Assessments in 5- to 30-Year-Olds After BNT162b2 Vaccination.

INTRODUCTION: Rare myocarditis and pericarditis cases have occurred in coronavirus disease 2019 (COVID-19) messenger RNA (mRNA) vaccine recipients. Troponin levels, a potential marker of myocardial injury, were assessed in healthy participants before and after BNT162b2 vaccination. METHODS: Vaccine-experienced 12- to 30-year-olds in phase 3 crossover C4591031 Substudy B (NCT04955626) who had two or three prior BNT162b2 30-µg doses were randomized to receive BNT162b2 30 µg followed by placebo, or placebo followed by BNT162b2 30 µg, 1 month apart. A participant subset, previously unvaccinated against COVID-19, in the phase 3 C4591007 study (NCT04816643) received up to three vaccinations (BNT162b2 10 µg or placebo [5- to 11-year-olds]) or open-label BNT162b2 30 µg (12- to 15-year-olds). Blood samples collected pre-vaccination, 4 days post-vaccination, and 1-month post-vaccination (C4591031 Substudy B only) were analyzed. Frequencies of elevated troponin I levels (male, > 35 ng/l; female, > 17 ng/l) were assessed. RESULTS: Percentages of 12- to 30-year-olds (n = 1485) in C4591031 Substudy B with elevated troponin levels following BNT162b2 or placebo receipt were 0.5% and 0.8% before vaccination, 0.7% and 1.0% at day 4, and 0.7% and 0.5% at 1 month, respectively. In Study C4591007 (n = 1265), elevated troponin I levels were observed in 0.2, 0.4, and 0.2% of 5- to 11-year-old BNT162b2 recipients at baseline and 4 days post-dose 2 and 3, respectively; corresponding values in 12- to 15-year-olds were 0.4, 0.4, and 0.7%. No 5- to 11-year-old placebo recipients had elevated troponin levels. No myocarditis or pericarditis cases or deaths were reported. CONCLUSIONS: Among 5- to < 30-year-olds in both studies, troponin levels were rarely elevated (≤ 1.0%) and similar before and post-vaccination; troponin levels were also similar between BNT162b2 and placebo in 12- to 30-year-old and 5- to 11-year-old recipients in the respective studies. No myocarditis or pericarditis cases were reported. These findings did not provide evidence that BNT162b2 causes troponin elevations. No utility of routine measurement of troponin levels in asymptomatic BNT162b2 recipients was identified.

[Translated article] Vancomycin powder in the prevention of infection in primary knee and hip arthroplasty: Case-control study with 1151 arthroplasties.

BACKGROUND AND OBJECTIVE: Vancomycin powder (VP) has been positively used in spinal surgery to reduce the rate of infections. Hardly any data have been published on hip and knee joint replacement surgery, and its usefulness is questioned. Our objective was to investigate the effectiveness of VP in reducing prosthetic infection and its possible complications. METHODS: Primary hip (THA) and knee (TKA) arthroplasties were reviewed, performed by five surgeons in one hospital centre, between 2017 and 2018. One gram of VP was used on the implant prior to surgical closure based on the surgeon's preferences. With a 5-year follow-up in which the infection rate and local complications were analysed. RESULTS: One thousand one hundred and fifty-one arthroplasties were performed, 748 were TKA and 403 were THA. Nine patients were diagnosed with prosthetic infection, of which five received VP and four did not (p=0.555). Likewise, another 15 patients suffered wound complications, of which 11 received VP and 4 did not (p=0.412). There were no differences, either, in the rest of the complications depending on the use or not of VP (p=0.101). Likewise, the number of patients who needed reintervention was similar (p=0.999). No systemic complications were detected due to the use of VP. CONCLUSIONS: It has not been possible to demonstrate that the use of VP reduces the rates of prosthetic infection in the hip and knee, so we cannot recommend its use.

Geographical differences in chronic obstructive pulmonary disease mortality trends by sex, Spain, 1980-2021.

BACKGROUND: To analyse changes in trends in mortality due to chronic obstructive pulmonary disease (COPD) in Spain by Autonomous Community (AC) and sex during the period 1980-2021.METHODS: Data on population and COPD death records (International Classification of Diseases, 10th edition, codes J40â-"J44 and J47) were retrieved from the National Institute of Statistics for the study period. Age-standardised mortality rates by AC and sex were assessed using joinpoint regression models.RESULTS: There were 562,668 deaths due to COPD (423,855 in men and 138,813 in women), with an average annual increase of 1.6%. COPD deaths in men increased in most ACs, except for Asturias (â-"0.5% per year). The Canary Islands (14% per year) and Madrid (6.5% per year) had the highest increases. In women, the figures show a wide range of values at the AC level (from a â-"1.4% decrease to 7.9% increase). Nationally, the sex ratio increased from 1980 to 2021. In men, six ACs showed a significant decrease, while in women only two ACs showed a significant decrease.CONCLUSION: A steady decrease in COPD mortality was observed in most ACs for men, while a different trend was observed in women in several ACs. Despite past and ongoing tobacco control initiatives, this condition remains a leading cause of death.

Vancomycin powder in the prevention of infection in primary knee and hip arthroplasty: Case-control study with 1151 arthroplasties. / Vancomicina en polvo en la prevención de la infección en artroplastia primaria de rodilla y cadera: estudio de casos y controles con 1.151 artroplastias.

BACKGROUND AND OBJECTIVE: Vancomycin powder (VP) has been positively used in spinal surgery to reduce the rate of infections. Hardly any data have been published on hip and knee joint replacement surgery, and its usefulness is questioned. Our objective was to investigate the effectiveness of VP in reducing prosthetic infection and its possible complications. METHODS: Primary hip (THA) and knee (TKA) arthroplasties were reviewed, performed by five surgeons in one hospital center, between 2017 and 2018. 1g of VP was used on the implant prior to surgical closure based on the surgeon's preferences. With a 5-year follow-up in which the infection rate and local complications were analyzed. RESULTS: One thousand one hundred and fifty one arthroplasties were performed, 748 were TKA and 403 were THA. Nine patients were diagnosed with prosthetic infection, of which five received VP and four did not (P=.555). Likewise, another 15 patients suffered wound complications, of which 11 received VP and 4 did not (P=.412). There were no differences, either, in the rest of the complications depending on the use or not of VP (P=.101). Likewise, the number of patients who needed reintervention was similar (P=.999). No systemic complications were detected due to the use of VP. CONCLUSIONS: It has not been possible to demonstrate that the use of VP reduces the rates of prosthetic infection in the hip and knee, so we cannot recommend its use.

A semi-automated microscopic image analysis method for scoring Ki-67 nuclear immunostaining

Nuclear proliferation marker MIB-1 (Ki-67) immunohistochemistry (IHC) is used to examine tumor cell proliferation. However, the diagnostic or prognostic value of the Ki-67 nuclear staining intensity and location, defined as nuclear gradient (NG), has not been assessed. This study examined the potential association between Ki-67 NG and cell cycle phases and its effect on the prognosis of pulmonary typical carcinoid (PTC) tumors. We propose a method for classifying the NG of Ki-67 during the cell cycle and compare the results between PTC, pulmonary adenocarcinoma (PAD), and breast ductal carcinoma (BDC). A literature review and objective analysis of IHC-stained paraffin sections were used to determine the Ki-67 labeling index and composed a stratification of the NG into NG1, NG2, and NG3/4 categories. A semi-automated image analysis protocol was established to determine the Ki-67 NG in PTC, PAD, and BDC. High intraobserver consistency and moderate interobserver agreement were achieved in the determination of Ki-67 NG in tumor specimens. NG1 and NG2 were lower in PTC than in PAD and BDC. Cox multivariate analysis of PTC after adjusting for age and number of metastatic lymph nodes showed that Ki-67 NG1 and NG2 significantly predicted clinical outcomes. The semi-automated method for quantification of Ki-67 nuclear immunostaining proposed in this study could become a valuable diagnostic and prognostic tool in PTC.

A new material to prevent urethral damage after implantation of artificial devices: an experimental study

ABSTRACT Objective To validate the application of the bacterial cellulose (BC) membrane as a protecting barrier to the urethra. Materials and Methods Forty female Wistar rats (four groups of 10): Group 1 (sham), the urethra was dissected as in previous groups and nothing applied around; Group 2, received a 0.7cm strip of the BC applied around the urethra just below the bladder neck; Group 3, received a silicon strip with the same dimensions as in group 2; Group 4, had a combination of 2 and 3 groups being the silicon strip applied over the cellulosic material. Half of the animals in each group were killed at 4 and 8 months. Bladder and urethra were fixed in formalin for histological analysis. Results Inflammatory infiltrates were more intense at 4 months at lymphonodes (80% Grade 2), statistically different in the group 2 compared with groups 1 (p=0.0044) and 3 (p=0.0154). At 8 months, all samples were classified as grade 1 indicating a less intense inflammatory reaction in all groups. In group 2, at 8 months, there was a reduction in epithelial thickness (30±1μm) when com-pared to groups 1 (p=0.0001) and 3 (p<0.0001). Angiogenesis was present in groups 2 and 4 and absent in group 3. In BC implant, at 4 and 8 months, it was significant when comparing groups 4 with 1 (p=0.0159). Conclusion BC membrane was well integrated to the urethral wall promoting tissue remodeling and strengthening based on morphometric and histological results and may be a future option to prevent urethral damage.

Levedura seca integral na alimentação de vacas lactantes / Dry yeast for lactating dairy cows

Os efeitos da inclusão de levedura seca de cana-de-açúcar (Saccharomyces cerevisiae) na dieta de vacas lactantes foram avaliados com base no fornecimento de quatro dietas constituídas de diferentes níveis de substituição do farelo de soja pela levedura de cana-de-açúcar (0,00; 0,33; 0,67 e 1,00kg/kg de MS). O delineamento utilizado foi o quadrado latino 4 × 4. Não houve efeito dos tratamentos sobre o consumo de MS, MO, EE, FDNi e NDT, porém ocorreu redução linear no consumo de PB e FDNcp e aumento do consumo de CNF. Diferenças estatísticas não foram verificadas para os coeficientes de digestibilidade aparente da MS, MO, PB, CNF e NDT; no entanto os coeficientes do FDNcp e do EE decresceram linearmente. Não foi observado efeito sobre a produção e eficiência de síntese microbiana, utilização dos componentes nitrogenados e produção e composição do leite. Conclui-se que a levedura seca pode substituir 100% o farelo de soja na alimentação de vacas lactantes.

The effects of the inclusion of sugar cane dry yeast (Saccharomyces cerevisiae) in dairy cows feeding were evaluated based on the provision of four different diets with increasing proportions of sugar cane dry yeast (0.00; 0.33; 0.67 and 1.00kg/kg of DM). The design adopted for this research was the latin square, 4 × 4. It was observed that the levels of yeast inclusion did not affect the intake of DM, OM, EE, NDFi and TDN. However, the CP and NDFcp intake decreased linearly, whereas the NFC intake increased linearly with the increasing levels of yeast. No effect on the digestibility coefficient of DM, OM, CP, NDFcp and NCF was observed. Nevertheless, the EE digestibility coefficient linearly decreased with the yeast inclusion in the experimental diets. There was no treatment effect on the production and efficiency of microbial synthesis, usage of nitrogenous components, as well as milk production and composition. It was concluded that the dried yeast can fully replace the soybean meal in diets of dairy cows.

Respuestas metabólicas inducidas por ejercicio físico de alta intensidad en mujeres sedentarias con glicemia basal alterada e hipercolesterolemia / Metabolic response to high intensity exercise training in sedentary hyperglycemic and hypercholesterolemic women

Background: High intensity training could be an effective way of improving health on individuals at high metabolic risk. Aim: To investigate the effects of a high intensity training intervention on metabolic-related markers in sedentary women at high metabolic risk. Material and Methods: Forty six sedentary women with a body mass index (BMI) over 25 kg/m² were assigned to four groups, according to their metabolic profile; hyperglycemia (H, n = 12), hyperglycemia/hypercholesterolemia (HH, n = 13), normoglycemia (N, n = 10) and normoglycemia/hypercholesterolemia (NH, n = 11). For 12 weeks and five days per week, subjects performed seven intervals of high intensity training (20 to 30 seconds) during a training session of 20 minutes. Anthropometric (body weight, body mass index (BMI), waist circumference) and metabolic variables (glucose, total cholesterol, LDL, HDL and TG) were measured at baseline, at 6 and 12 weeks of intervention. Results: BMI and waist circumference decreased significantly after 12 weeks of intervention. Similarly, glucose decreased significantly after 12 weeks of intervention in all groups. The reduction was of higher magnitude in those groups with hyperglycemia (H = -16%, HH = -22%, N = -7,5%, NH = -9,6%). However, lipid profile (TG, total cholesterol, LDL and HDL) improved significantly only in the hypercholesterolemic groups. Conclusions: Physical activity programs incorporating high intensity training can improve glucose and lipid profile in women with metabolic disorders. Moreover, this benefit is greatest in those individuals with highest metabolic burden.

Substituição do feno de tifton pela casca de mamona na dieta de ovinos: componentes não-carcaça / Replacement of tifton hay with castor bean hulls in sheep diet: non carcass components

Avaliou-se o efeito da substituição do feno de capim tifton pela casca de mamona, 0, 33, 66 e 100% na dieta de ovinos, sobre o peso e rendimento dos componentes não constituintes da carcaça e o rendimento de buchada e panelada. Vinte e oito animais não castrados, com sete meses de idade e peso inicial médio de 19,5±2,45kg, foram distribuídos em delineamento de blocos ao acaso, com quatro tratamentos e sete repetições, e abatidos após 70 dias de confinamento. Houve decréscimo linear nos pesos da língua (0,084 a 0,079kg), traqueia+pulmão (0,50 a 0,42kg), coração (0,14 a 0,12kg), timo (0,035 a 0,025kg), rins (0,100 a 0,088kg) e peso total dos órgãos (2,14 a 1,90kg) de acordo com o percentual de substituição. O mesmo comportamento foi verificado para os pesos do esôfago (0,051 a 0,045kg), rúmen-retículo (0,73 a 0,64kg), intestino delgado (0,78 a 0,64kg) e total de vísceras (2,17 a 1,92kg). Para os pesos da cabeça (1,99 a 1,66kg), patas (0,86 a 0,73kg), pele (2,49 a 1,99kg), sangue (1,36 a 1,19kg) e total de subprodutos (7,12 a 5,96kg) verificou-se efeito linear negativo de acordo com a substituição. O mesmo foi observado para os pesos de buchada (4,71 a 4,22kg) e panelada (7,55 a 6,61kg). A casca de mamona não deve substituir o feno de capim tifton na dieta de ovinos por causar decréscimo no peso de órgãos, vísceras e subprodutos e influenciar negativamente os pesos e rendimentos de buchada e panelada.

The effects of replacing tifton hay with 0, 33, 66 and 100% of castor bean hulls in the diet of sheep on the non-carcass components of weight and dressing and on the "buchada" and "panelada" yield were evaluated in this research. Twenty-eight non castrated sheep averaging seven months of age and with initial weight of 19.5±2.45kg were assigned to a randomized block design with four treatments and seven replicates, and were slaughtered after 70 days of confinement. A linear decrease of tongue (0.084 to 0.079kg), trachea + lungs (0.50 to 0.42kg), heart (0.14 to 0.12kg), thymus (0.035 to 0.025kg), kidneys (0.100 to 0.088kg) weights and total weight of organs (2.14 to 1.90kg) was observed. The same behavior was observed for the esophagus (0.051 to 0.045kg), rumen-reticulum (0.73 to 0.64kg), small intestine (0.78 to 0.64kg) and total viscera (2.17 to 1.92kg) weights. There was a linear effect for the head (1.99 to 1.66kg), feet (0.86 to 0.73kg), skin (2.49 to 1.99kg), blood (1.36 to 1.19kg) and total by-products (7.12 to 5.96kg) weights according to the treatments and also for "buchada" (4.71 to 4.22kg) and "panelada" (7.55 to 6.61kg) weights. Castor bean hulls should not replace tifton hay because it decreases the weight of organs, viscera and by-products and adversely affects the "buchada" and "panelada" weight and dressing.

Production of L1 protein from different types of HPV in Pichia pastoris using an integrative vector

Human papillomavirus (HPV) infection is the most common sexually transmitted disease in the world and is related to the etiology of cervical cancer. The most common high-risk HPV types are 16 and 18; however, the second most prevalent type in the Midwestern region of Brazil is HPV-33. New vaccine strategies against HPV have shown that virus-like particles (VLP) of the major capsid protein (L1) induce efficient production of antibodies, which confer protection against the same viral type. The methylotrophic yeast Pichia pastoris is an efficient and inexpensive expression system for the production of high levels of heterologous proteins stably using a wild-type gene in combination with an integrative vector. It was recently demonstrated that P. pastoris can produce the HPV-16 L1 protein by using an episomal vector associated with the optimized L1 gene. However, the use of an episomal vector is not appropriate for protein production on an industrial scale. In the present study, the vectors were integrated into the Pichia genome and the results were positive for L1 gene transcription and protein production, both intracellularly and in the extracellular environment. Despite the great potential for expression by the P. pastoris system, our results suggest a low yield of L1 recombinant protein, which, however, does not make this system unworkable. The achievement of stable clones containing the expression cassettes integrated in the genome may permit optimizations that could enable the establishment of a platform for the production of VLP-based vaccines.

Avaliação da atividade antioxidante do extrato aquoso de Lafoensia pacari A. ST-HIL. em emulsão não-iônica / Evaluation of antioxidant activity of aqueous extract of Lafoensia pacari A. ST.-HIL. in non-ionic emulsion

O envelhecimento cutâneo está relacionado com a ação de radicais livres que, por consequência da senescência, acabam estando em maior quantidade e logo as defesas naturais não conseguem eliminá-los. O objetivo deste trabalho foi desenvolver uma emulsão não-iônica com adição do extrato aquoso das entrecascas de Lafoensia pacari A. St.-Hil. e avaliar sua estabilidade físico-química, assim como a atividade antioxidante. O extrato aquoso foi preparado sob refluxo na proporção 1:25. A dosagem de polifenóis totais se deu pelo método Folin-Ciocalteau, com ácido tânico como padrão. Foram incorporados à emulsão não-iônica BHT e ácido ascórbico, ambos empregados como substâncias de referência e o extrato aquoso na concentração de 1%. A análise da atividade antioxidante foi realizada pelo método do fosfomolibdênio, onde cada amostra foi diluída a concentração de 400 miug/mL. A incorporação de extrato aquoso de L. pacari não influenciou de maneira significativa na estabilidade do creme em relação ao tempo de análise. A quantificação de polifenóis mostrou que a extração aquosa à quente não foi um método eficiente, resultando em um EAT (equivalente de ácido tânico) de 1,0328 mg/mL. Proporcional à concentração de polifenóis, a atividade antioxidante foi baixa, não ultrapassando 5,0% de atividade em relação ao ácido ascórbico.

Cutaneous aging is related to the action of free radicals, which appear in greater quantity as a consequence of senescence, so that natural defenses cannot eliminate them. The aim of this study was to develop a non-ionic emulsion containing an aqueous extract of the inner bark of Lafoensia pacari A. St.-Hil. and assess its physicochemical stability, as well as its antioxidant activity. The aqueous extract was prepared under reflux with a 1:25 ratio of bark:distilled water (w/v). The total polyphenol content was determined by the Folin-Ciocalteau method, with tannic acid as standard. BHT, ascorbic acid and the aqueous extract were incorporated into the non-ionic emulsion, each of them at a concentration of 1%, the first two being. used as reference substances. Antioxidant activity was analyzed by the phosphomolybdenum method, each emulsion being first diluted in water to 400 ìg / mL. The addition of aqueous extract of L. pacari did not significantly affect the stability of the cream over the period of analysis. The content of polyphenols showed that hot aqueous extraction was not an efficient method, resulting in a tannic acid equivalent (TAE) of 1.0328 mg/mL in the crude extract. Proportional to the polyphenol concentration, the antioxidant activity was also low, not exceeding 5.0% of the activity of the ascorbic acid.

Atitudes em relação à Bioestatística de discentes e docentes da Faculdade de Ciências Farmacêuticas de Araraquara-UNESP / Attitudes towards Biostatistics of students and teachers at the Faculty of Pharmaceutical Sciences of Araraquara-UNESP

O objetivo deste estudo foi verificar a atitude de discentes e docentes da Faculdade de Ciências Farmacêuticas de Araraquara (UNESP) em relação à Bioestatística. Como instrumento de medida, utilizou-se a Escala de Atitudes em Relação à Estatística (EAE). A reprodutibilidade da Escala foi estimada pela estatística Kappa (κ) com ponderação linear e sua consistência interna pelo Coeficiente alfa-Cronbach (α). Os indivíduos foram agrupados segundo sua atitude em relação à Estatística em positiva e negativa; posteriormente, estudou-se sua associação com as variáveis de interesse pelo teste de quiquadrado (x2) ao nível de significância de 5%. A amostra foi composta por 272 alunos de graduação, 83 de pósgraduação e 24 docentes, sendo predominantemente feminina (78,2%). Entre os estudantes, 67,5% participaram do programa de iniciação científica. A reprodutibilidade e a consistência interna da Escala foram adequadas (κ=0,7093; α=0,9334). A maior parte da amostra (74,4%) apresentou atitude positiva frente à Estatística. Houve associação entre a atitude e a atividade funcional (p=0,0204), série cursada (p=0,0316) e desempenho (p=0,0002). Assim, concluise que a maioria dos participantes apresentou atitude positiva em relação à Bioestatística, sendo que os estudantes de graduação e aqueles que relataram bom desempenho em Bioestatística apresentaram proporção significativamente maior de atitude positiva em relação aos demais.

In this study, the attitudes of students and teachers at the Faculty of Pharmaceutical Sciences of Araraquara (UNESP University) towards Biostatistics were assessed. The Survey of Attitudes Toward Statistics (SATS) scale was used as the measuring instrument. The reproducibility of the scale was estimated by Cohen's Kappa (k) coefficient with linear weighting and its internal consistency by Cronbach's alpha coefficient (α). The individuals were first placed in two groups, according to their positive or negative attitude toward Statistics; then, the association of their attitude with the variables of interest was tested by the chi-squared (χ2) test at a significance level of 5%. The sample consisted of 272 undergraduate students, 83 graduate students and 24 teachers, predominantly female (78.2%). Among the students, 67.5% participated in the scientific research initial training program. Reproducibility and internal consistency of the scale were adequate ((κ=0.7093; α=0.9334). Most of the subjects (74.4%) had a positive attitude toward Statistics. Significant association was found between attitude and functional activity (p=0.0204), the course taken (p=0.0316) and effort (p=0.0002). Thus, it was concluded that the great majority of the participants had a positiveattitude towards Biostatistics and that undergraduate students and those who reported good performance in Biostatistics showed a significantly higher proportion of positive attitude than the other students.

Confiabilidade de um formulário para diagnóstico da severidade da disfunção temporomandibular / Reliability of a questionnaire for diagnosing the severity of temporomandibular disorder

CONTEXTUALIZAÇÃO: Disfunção temporomandibular (DTM) é um termo coletivo que engloba vários problemas clínicos envolvendo a musculatura da mastigação, as articulações temporomandibulares (ATM) e suas estruturas associadas, com alta prevalência nas populações. OBJETIVOS: Sabendo-se que estudos brasileiros vêm utilizando o instrumento proposto por Da Fonseca et al. (1994) para diagnóstico da severidade desta disfunção, realizou-se este estudo com o objetivo de verificar e estimar a consistência interna e a reprodutibilidade do mesmo. MÉTODOS: O delineamento amostral adotado foi o probabilístico, e participaram 1230 indivíduos moradores da cidade de Ribeirão Preto (SP), maiores de 18 anos de idade. As entrevistas foram realizadas por um único entrevistador por meio de ligações telefônicas. Para estudo da consistência interna, calculou-se o Coeficiente de Kuder-Richardson (kr-20) e para estimar a reprodutibilidade, utilizou-se a estatística Kappa (κ). RESULTADOS: A consistência interna do formulário foi de 0,5594, apontando para uma validação abaixo do desejado. Observou-se maior contribuição das questões 1, 2, 3, 6 e 7 para o coeficiente kr-20 total e maior consistência do instrumento quando composto apenas pelas mesmas (0,7044). Observou-se reprodutibilidade "Boa" e "Ótima" para as questões. CONCLUSÕES: Frente ao exposto, sugere-se que o formulário proposto por Da Fonseca et al. (1994) seja adaptado, ficando composto apenas pelas questões 1, 2, 3, 6 e 7 da versão inicial, colaborando, assim, para aumento da confiabilidade do instrumento. Deve-se ressaltar ainda a necessidade da realização de estudos de validade para assegurar adequadas características psicométricas à nova versão do instrumento.

BACKGROUND: Temporomandibular disorder (TMD) is a collective term that encompasses many clinical problems involving the masticatory muscles, temporomandibular joints (TMJ) and associated structures and it has high prevalence among populations. OBJECTIVES: Because Brazilian studies have used the instrument proposed by Da Fonseca et al. (1994) to diagnose the severity of TMD, this study was conducted to investigate and estimate the internal consistency and reproducibility of this method. METHODS: We used a probability sampling design to select 1230 participants over the age of 18 years who were living in the city of Ribeirão Preto, SP, Brazil. The interviews were conducted by a single interviewer over the phone. The internal consistency was analyzed by calculating the Kuder-Richardson coefficient (kr-20), and kappa statistics (κ) were used to estimate the reproducibility. RESULTS: The internal consistency of the questionnaire was 0.5594, thus indicating that validation was lower than desired. Questions 1, 2, 3, 6 and 7 had greater contribution towards the total kr-20 coefficient, and the consistency of the instrument was higher when it was composed only of these questions (0.7044). "Good" and "Excellent" reproducibility was observed for these same questions. CONCLUSIONS: Based on these data, it is suggested that the questionnaire proposed by Da Fonseca et al. (1994) should be adapted to include only questions 1, 2, 3, 6 and 7 of the initial version. This would help improve the reliability of the instrument. The need for validation studies must also be emphasized to ensure that the new version of the instrument has adequate psychometric characteristics.