Measurement of serum estrogen and estrogen metabolites in pre- and postmenopausal women with osteoarthritis using high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry

Although 17β-estradiol (E2) deficiency has been linked to the development of osteoarthritis (OA) in middle-aged women, there are few studies relating other estrogens and estrogen metabolites (EMs) to this condition. We developed a high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (HPLC-ESI-MS/MS) method to measure the levels of six EMs (i.e., estrone, E2, estriol, 2-hydroxyestrone, 2-hydroxyestradiol, and 16a-hydroxyestrone) in healthy pre- and postmenopausal women and women with OA. This method had a precision ranging from 1.1 to 3.1% and a detection limit ranging from 10 to 15 pg. Compared to healthy women, serum-free E2 was lower in the luteal and postmenopausal phases in women with OA, and total serum E2 was lower in postmenopausal women with OA. Moreover, compared to healthy women, total serum 2-hydroxyestradiol was higher in postmenopausal women with OA and total serum 2-hydroxyestrone was lower in both the luteal and follicular phases in women with OA. In conclusion, our HPLC-ESI-MS/MS method allowed the measurement of multiple biochemical targets in a single assay, and, given its increased cost-effectiveness, simplicity, and speed relative to previous methods, this method is suitable for clinical studies.

Hyaluronidases and hyaluronan synthases expression is inversely correlated with malignancy in lung/bronchial pre-neoplastic and neoplastic lesions, affecting prognosis

We collected a series of 136 lung/bronchial and 56 matched lung parenchyma tissue samples from patients who underwent lung/bronchial biopsies and presented invasive carcinoma after lung surgery. The lung/bronchial samples included basal cell hyperplasia, squamous metaplasia, moderate dysplasia, adenomatous hyperplasia, severe dysplasia, squamous cell carcinoma and adenocarcinoma. Matched lung parenchyma tissue samples included 25 squamous cell carcinomas and 31 adenocarcinomas. Immunohistochemistry was performed to analyze for the distribution of hyaluronidase (Hyal)-1 and −3, and hyaluronan synthases (HAS)-1, −2, and −3. Hyal-1 showed significantly higher expression in basal cell hyperplasia than in moderate dysplasia (P=0.01), atypical adenomatous hyperplasia (P=0.0001), or severe dysplasia (P=0.03). Lower expression of Hyal-3 was found in atypical adenomatous hyperplasia than in basal cell hyperplasia (P=0.01) or moderate dysplasia (P=0.02). HAS-2 was significantly higher in severe dysplasia (P=0.002) and in squamous metaplasia (P=0.04) compared with basal cell hyperplasia. HAS-3 was significantly expressed in basal cell hyperplasia compared with atypical adenomatous hyperplasia (P=0.05) and severe dysplasia (P=0.02). Lower expression of HAS-3 was found in severe dysplasia compared with squamous metaplasia (P=0.01) and moderate dysplasia (P=0.01). Epithelial Hyal-1 and −3 and HAS-1, −2, and −3 expressions were significantly higher in pre-neoplastic lesions than in neoplastic lesions. Comparative Cox multivariate analysis controlled by N stage and histologic tumor type showed that patients with high HAS-3 expression in pre-neoplastic cells obtained by lung/bronchial biopsy presented a significantly higher risk of death (HR=1.19; P=0.04). We concluded that localization of Hyal and HAS in lung/bronchial pre-neoplastic and neoplastic lesions was inversely related to malignancy, which implied that visualizing these factors could be a useful diagnostic procedure for suspected lung cancer. Finalizing this conclusion will require a wider study in a randomized and prospective trial.

Concomitant stress potentiates the preference for, and consumption of, ethanol induced by chronic pre-exposure to ethanol

Ethanol abuse is linked to several acute and chronic injuries that can lead to health problems. Ethanol addiction is one of the most severe diseases linked to the abuse of this drug. Symptoms of ethanol addiction include compulsive substance intake and withdrawal syndrome. Stress exposure has an important role in addictive behavior for many drugs of abuse (including ethanol), but the consequences of stress and ethanol in the organism when these factors are concomitant results in a complex interaction. We investigated the effects of concomitant, chronic administration of ethanol and stress exposure on the withdrawal and consumption of, as well as the preference for, ethanol in mice. Male Swiss mice (30–35 g, 8-10 per group) were exposed to an ethanol liquid diet as the only source of food for 15 days. In the final 5 days, they were exposed to forced swimming stress. Twelve hours after removal of the ethanol liquid diet, animals were evaluated for ethanol withdrawal by measuring anxiety-related behaviors and locomotor activity. Twenty-four hours after evaluation of ethanol withdrawal, they were evaluated for voluntary consumption of ethanol in a “three-bottle choice” paradigm. Mice exposed to chronic consumption of ethanol had decreased locomotor activity during withdrawal. Contrary to our expectations, a concomitant forced swimming stress did not aggravate ethanol withdrawal. Nevertheless, simultaneous ethanol administration and stress exposure increased voluntary consumption of ethanol, mainly solutions containing high concentrations of ethanol. These results showed that stressful situations during ethanol intake may aggravate specific addiction-related behaviors.

Lyophilized allografts without pre-treatment with glutaraldehyde are more suitable than cryopreserved allografts for pulmonary artery reconstruction

Various methods are available for preservation of vascular grafts for pulmonary artery (PA) replacement. Lyophilization and cryopreservation reduce antigenicity and prevent thrombosis and calcification in vascular grafts, so both methods can be used to obtain vascular bioprostheses. We evaluated the hemodynamic, gasometric, imaging, and macroscopic and microscopic findings produced by PA reconstruction with lyophilized (LyoPA) grafts and cryopreserved (CryoPA) grafts in dogs. Eighteen healthy crossbred adult dogs of both sexes weighing between 18 and 20 kg were used and divided into three groups of six: group I, PA section and reanastomosis; group II, PA resection and reconstruction with LyoPA allograft; group III, PA resection and reconstruction with CryoPA allograft. Dogs were evaluated 4 weeks after surgery, and the status of the graft and vascular anastomosis were examined macroscopically and microscopically. No clinical, radiologic, or blood-gas abnormalities were observed during the study. The mean pulmonary artery pressure (MPAP) in group III increased significantly at the end of the study compared with baseline (P=0.02) and final [P=0.007, two-way repeat-measures analysis of variance (RM ANOVA)] values. Pulmonary vascular resistance of groups II and III increased immediately after reperfusion and also at the end of the study compared to baseline. The increase shown by group III vs group I was significant only if compared with after surgery and study end (P=0.016 and P=0.005, respectively, two-way RM ANOVA). Microscopically, permeability was reduced by ≤75% in group III. In conclusion, substitution of PAs with LyoPA grafts is technically feasible and clinically promising.

Monolithic zirconium dioxide as a full contour restorative material: With special emphasis on the optical and mechanical properties

Full contour monolithic zirconia restorations have shown an increased popularity in the dental field over the recent years, owing to its mechanical and acceptable optical properties. However, many features of the restoration are yet to be researched and supported by clinical studies to confirm its place among the other indirect restorative materials This series of in vitro studies aimed at evaluating and comparing the optical and mechanical properties, light cure irradiance, and cement polymerization of multiple monolithic zirconia material at variable thicknesses, environments, treatments, and stabilization. Five different monolithic zirconia materials, four of which were partially stabilized and one fully stabilized were investigated. The optical properties in terms of surface gloss, translucency parameter, and contrast ratio were determined via a reflection spectrophotometer at variable thicknesses, coloring, sintering method, and after immersion in an acidic environment. Light cure irradiance and radiant exposure were quantified through the specimens at variable thicknesses and the degree of conversion of two dual-cure cements was determined via Fourier Transform Infrared spectroscopy. Bi-axial flexural strength was evaluated to compare between the partially and fully stabilized zirconia prepared using different coloring and sintering methods. Surface characterization was performed using a scanning electron microscope and a spinning disk confocal microscope. The surface gloss and translucency of the zirconia investigated were brand and thickness dependent with the translucency values decreasing as the thickness increased. Staining decreased the translucency of the zirconia and enhanced surface gloss as well as the flexural strength of the fully stabilized zirconia but had no effect on partially stabilized zirconia. Immersion in a corrosive acid increased surface gloss and decreased the translucency of some zirconia brands. Zirconia thickness was inversely related to the amount of light irradiance, radiant exposure, and degree of monomer conversion. Type of sintering furnace had no effect on the optical and mechanical properties of zirconia. Monolithic zirconia maybe classified as a semi-translucent material that is well influenced by the thickness, limiting its use in the esthetic zones. Conventional acid-base reaction, autopolymerizing and dual-cure cements are recommended for its cementation. Its desirable mechanical properties give it a high potential as a restoration for posterior teeth. However, close monitoring with controlled clinical studies must be determined before any definite clinical recommendations can be drawn.

In vitro protein digestibility of enzymatically pre-treated bean (Phaseolus vulgaris L.) flour using commercial protease and Bacillus sp. protease

The common bean (Phaseolus vulgaris L.) is a staple food in the Brazilian diet and represents the major source of dietary protein and other micronutrients and minerals. Despite the considerable protein concentration in beans, the food is considered of low biological value when compared to animal proteins and other plant protein sources. To improve the availability of protein in beans, enzymatic treatments were performed in four cultivars (ON, OPNS, TAL and VC3). The approach was a completely randomized design with four replicates. We used a 4 × 3 factorial arrangement (four cultivars and three treatments: treatment 1-addition of commercial protease (Trypsin 250, Difco), treatment 2-addition of protease from Bacillus sp., and treatment 3:-control without enzyme addition). The enzyme: substrate ratio was 5% w/w (amount of enzyme per total protein in bean flour). The approach was a completely randomized design with four replicates. A 4 × 3 factorial arrangement (four cultivars and three treatments, the same as those mentioned above) was used. The concentration of total protein (g.100 g-1 of dry matter) in the samples ranged from 16.94 to 18.06%, while the concentration of total phenolics was between 0.78 and 1.12% (g Eq. tannic acid.100 g-1 dry matter). The in vitro protein digestibility of enzymatically untreated bean flour (control) ranged from 47.30 to 56.17% based on the digestibility of casein. Concentrations of P, K, Ca, Mg, and Zn observed in the four cultivars tested were within the average values available in the literature. Treatment 2 with protease from Bacillus sp. induced decreases in the levels of Cu and Mn. The average Fe content increased in all bean flour samples when treated with proteases, reaching a maximum increase of 102% in the TAL flour treated with protease from Bacillus sp. The digestibility of all beans tested was significantly increased (p < 0.05) after the enzyme treatment. The greatest change was observed in the OPNS cultivar treated with protease from Bacillus sp., which increased its digestibility from 54.4% (control treatment) to 81.6%.

Pre-treatment of thermoduric spores in CO2 modified atmosphere and their survivability during food extrusion

The aim of this experiment was to evaluate how susceptible spores become to mechanical damage during food extrusion after being submitted to CO2. B. stearothermophilus spores sowed to corn and soy mix were submitted to 99% CO2 for 10 days and extruded in a single-screw extruder. The treatments were: T1 - spore-containing samples, extruded at screw rotational speed of 65 rpm and barrel wall temperature of 80 °C; T2 - as T1, except for screw rotational speed of 150 rpm; and T3 - as T2, except that samples were submitted to the modified atmosphere. The results for cell viability, minimum and maximum residence times, and static pressure were T1 - 19.90 ± 3.24%, 123.3 ± 14.50 seconds; 203.3 ± 14.05 seconds; 2.217 ± 62 kPa; T2 - 21.42 ± 8.24%, 70.00 ± 5.77 seconds; 170.00 ± 4.67 seconds; 2.310 ± 107 kPa; and T3 - 11.06 ± 2.46%, 86.00 ± 7.23 seconds; 186.00 ± 7.50 seconds; 2.403 ± 93 kPa, respectively. It was concluded that the extrusion process did reduce the cell count. However, screw rotational speed variation or CO2 pre-treatment did not affect cell viability.

Microstructural changes of frozen strawberries submitted to pre-treatments with additives and vacuum impregnation

Strawberries were submitted to freezing after pre-treatments with hydrocolloid and calcium salts (pectin and calcium chloride) at different concentrations, in the attempt to establish a correlation of the effects of these substances and their processing, on the physical and microstructural characteristics of fruits after thawing. Strawberry halves were submitted to impregnation with controlled vacuum pressure of 84.4, 50.5 and 16.6 kPa; comprising pectin at concentrations of 0, 1.5 and 3%; with the addition of calcium chloride at concentrations of 0, 3 and 6%; and glucose at 20%, for 4 hours. Measurements were made of the total soluble solid contents, cellular fluid loss, texture and viscosity of the solution, before and after the freezing/thawing. Images of the tissue cuts during the freezing, in function of time, were taken in an optic microscope coupled to a cold-stage and controlled temperature system, where the reduction of the cellular area was quantified using an image analyzing software. The pectin concentration had an influence on and demonstrated a potential for protection of the frozen tissue samples. The photomicrographs showed that the loss of cellular fluid occurs during the growth of ice formed in the intercellular spaces and it is retarded through treatments with high pectin concentrations.

Saline distribution during multicomponent salting in pre-cooked quail eggs

The relationship of NaCl with problems of arterial hypertension has led to a reduction in the levels of this salt in food production. KCl has been used as a partial substitute for NaCl since it cannot be completely substituted without affecting the acceptability of the end product. In this study, the diffusion that occurs during quail egg salting in static and stirred brine was simulated. The mathematical model used was based on a generalization of the Fick's 2nd law, and the COMSOL Multiphysics software was used to simulate the diffusion in the NaCl-KCl-water system. The deviations in the simulated data and experimental data were 2.50% for NaCl and 6.98% for KCl in static brine, while in the stirred brine they were 3.48% for NaCl and 4.72% for KCl. The simulation results presented good agreement with the experimental values and validated the predictive capacity of the model.

Effect of pre and post-slaughter processes on meat characteristics of Santa Ines ewes discarded due to age

The effects of body condition recovery (BC), carcass electrical stimulation (ES), aging time (AT 7 - 14 days), and calcium chloride injection on the meat characteristics of Santa Inês ewes (±5 years old) slaughtered immediately after weaning or after the body condition recovery period were studied. The carcass temperature, pH, shear force (SF), cooking loss (CL), meat color (L*, a*, b*), and meat tenderness were evaluated. A completely randomized design in a 2 × 2 × 2 × 3 (BC × ES × CaCl2 × AT) factorial arrangement was used, and the sensory tenderness data were analyzed using the table of Minimum Number of Correct Answers for the Duo-Trio test. The body condition recovery reduces the shear force in 8%, increasing their tenderness. Electrical stimulation reduced the shear force (24%) and did not change the other parameters. The aging time (7 or 14 days) decreased the shear force (18-26%), effect that was enhanced by electrical stimulation, and it darkened the meat reducing lightness (L*) and increasing yellowness (b*). The treatment with CaCl2 was the most effective in tenderizing meat by reducing the shear force ( 35%); increasing the cooking loss (4.5%); and increasing L* and b* lightening the meat. The sensory evaluation of tenderness corroborates the findings of the experimental evaluation regarding the effect of the treatment with CaCl2 on the meat quality improvement. It was concluded that the treatments improve meat characteristics achieving better results when applied together.

Characterization of pre-gelatinized rice and bean flour

The objective of this study was to develop a pre-gelatinized flour using a mixture of broken rice and split beans by thermoplastic extrusion, and to evaluate the physicochemical, nutritional, and technological quality of the final product. The extrusion parameters were maintained using three heating zones with temperatures of 30 ºC, 40 ºC, and 70 ºC; screw speed of 177 rpm; feed rate of 257 g/m, and circular matrix of 3.85 mm. The following characterization analyses were performed: physicochemical, nutritional, water absorption index (WAI), water solubility index (WSI), and paste viscosity. The pre-gelatinized rice and bean flour had an intermediate value of WAI, 7.51 g/g, and high WSI value, 24.61%. Regarding proteins, it was verified an average content of 12.9% in the final product. The amino acid contents found in the pre-gelatinized flour indicate that the mixture has the essential amino acids. It was also found that the pre-gelatinized flour supplies more than 60% of the essential amino acids recommended for children aged one to three years old. The gelatinized flour composed of broken rice and split beans is an alternative to the use of these by-products of the manufacture process of rice and beans to obtain a product with viable technological characteristics and high nutritional value.