Antonio Silverio Cabrita

Assessment of markers of bone formation under controlled environmental factors and their correlation with serum minerals in adult sheep as a model for orthopaedic research

Summary Eighteen healthy skeletally mature (3 years old) ewes, with an average weight of 45 kg, of the Portuguese Churra da Terra Quente breed were used to evaluate the normal values of total and bone-specific isoform of alkaline phosphatase serum activities (ALP and BALP, respectively) and serum osteocalcin (OC) and their correlation with the serum minerals - calcium (Ca), phosphorus (P), magnesium (Mg) and ionized calcium (Ca2+). The sheep were maintained under controlled environmental conditions (constant diurnal photoperiod cycle and identical husbandry and feeding) for six weeks before the collection of the blood samples. The measurement of the total ALP and serum minerals was performed with automated biochemistry analysers using the BioMérieux® kits, the serum electrolyte Ca2+ Diametrics Medical, Inc® specific cassettes and the BALP and OC METRATM kits from QUIDEL® Corporation. The mean ± standard deviation values obtained were: total ALP 90.17 ± 85.72 U/L, BALP 15.0 ± 5.44 U/L, ratio BALP/ total ALP 29.28 ± 24.22, OC 13.02 ± 1.87 ng/mL, Ca 2.57 ± 0.37 mmol/L, P 2.13 ± 0.42 mmol/L, Mg 1.04 ± 0.13 mmol/L, Ca2+ 1.29 ± 0.04 mmol/L. Significant correlations were observed between the total ALP and Ca (r = 0.5939; P = 0.05) and OC and Ca (r = 0.5706; P = 0.05). Reference to the serum values of bone turnover parameters in sheep could be of great value in research and could provide complementary non-invasive information on the bone healing process, particularly with regard to obtaining an early prognosis of fracture healing.

Haematological and biochemical parameters in Churra-da-Terra-Quente ewes from the northeast of Portugal

Hematological and biochemical parameters, including plasma electrolytes and thyroid hormones, were determined in 73 clinically healthy Churra-da-Terra-Quente ewes, a typical breed from the northeast of Portugal. The hemogram values were: erythrocytes 9.8±1.5×1012/L; haemoglobin 118.1±19.1g/L; haematocrit 40.8±5.9%; leukocytes 5.7±1.8×109/L; and platelets 544.3±177.2×109/L. The thrombin time was 17.3±1.7 seconds. The values of biochemical parameters were: total protein 76.4±6.1g/L; glucose 2.87±0.60mmol/L; total cholesterol 1.65±0.33mmol/L; aspartate aminotransferase 155.9±49.2U/L; alanine aminotransferase 23.2±9.6U/L; γ-glutamyl transferase 48.0±18.7U/L; total alkaline phosphatase 121.6±76.1U/L; glutamate dehydrogenase 6.4±3.7U/L; urea 7.32±2.22mmol/L; creatinine 123.0±54.1μmol/L; total calcium 2.53±0.25mmol/L; phosphorus 2.10±0.46mmol/L; magnesium 1.01±0.09mmol/L; sodium 152.04±3.65mmol/L; potassium 4.7±0.4mmol/L; ionized calcium 1.32±0.07mmol/L; total thyroxine 111.75±42.29nmol/L; total triiodothyronine 1.01±0.28nmol/L; free T4 11.93±1.78pmol/L; free T3 4.22±1.33pmol/L; and thyroid-stimulating hormone 0.18±0.19μIU/mL. Although differences among the Churra-da-Terra-Quente breed and other breeds may occur, the hematological and biochemical parameters, plasma electrolytes, and thyroid hormones, for this indigenous breed, were generally situated within the reference intervals previously reported for sheep.

Effects on gastric mucosa induced by dental bleaching – an experimental study with 6% hydrogen peroxide in rats

The value of aesthetic dentistry has precipitated several developments in the investigation of dental materials related to this field. The free marketing of these products is a problem and it is subject to various interpretations regarding its legality. There are several techniques for tooth whitening, the most used one being the external bleaching. It is the later version of such technique that poses the greatest danger of ingesting the product. The present study analysed the systemic effect of these products when they are swallowed. Objective This experimental study aimed to observe the effects of a tooth whitening product, whose active agent is 6% hydrogen peroxide, on the gastric mucosa of healthy and non-tumour gastric pathology animals. Material and Methods Fifty Wistar-Han rats were used and then distributed into 5 groups, one for control and four test groups in which the bleaching product was administered in animals with and without non-tumour gastric pathology (induced by the administration of 1 sample of 50% ethanol and 5% of drinking water during 6 days) at different times of study by gavage. There was a decrease in body weight in animals of groups handled during the study period, which was most pronounced in IV and VA groups. Changes in spleen weight relative to body weight revealed no statistically significant changes. An analysis of the frequency was performed on the results of macroscopic observation of the gastric mucosa. Results The gastric mucosa revealed lesions in all manipulated groups, being more frequent in groups III and IV. It appears that there is a synergism when using hydrogen peroxide and 50% ethanol in the same group. Conclusion Therefore, it seems that there are some signs of toxicity 3 to 4 days after administration of 6% hydrogen peroxide. The prescription of these therapies must be controlled by the clinician and the risks must be minimized.

Early mortality after neonatal surgery: analysis of risk factors in an optimized health care system for the surgical newborn

OBJECTIVE Anesthetic and operative interventions in neonates remain hazardous procedures, given the vulnerability of the patients in this pediatric population. The aim was to determine the preoperative and intraoperative factors associated with 30-day post-operative mortality and describe mortality outcomes following neonatal surgery under general anesthesia in our center. METHODS Infants less than 28 days of age who underwent general anesthesia for surgery during an 11-year period (2000 - 2010) in our tertiary care pediatric center were retrospectively identified using the pediatric intensive care unit database. Multiple logistic regression was used to identify independent preoperative and intraoperative factors associated with 30-day post-operative mortality. RESULTS Of the 437 infants in the study (median gestational age at birth 37 weeks, median birth weight 2,760 grams), 28 (6.4%) patients died before hospital discharge. Of these, 22 patients died within the first post-operative month. Logistic regression analysis showed increased odds of 30-day post-operative mortality among patients who presented American Society of Anesthesiologists physical status (ASA) score 3 or above (odds ratio 19.268; 95%CI 2.523 - 147.132) and surgery for necrotizing enterocolitis/gastrointestinal perforation (OR 5.291; 95%CI 1.962 - 14.266), compared to those who did not. CONCLUSION The overall in-hospital mortality of 6.4% is within the prevalence reported for developed countries. Establishing ASA score 3 or above and necrotizing enterocolitis/gastrointestinal perforation as independent risk factors for early mortality in neonatal surgery may help clinicians to more adequately manage this high risk population.

Evaluation of the effect of acupuncture on hand pain, functional deficits and health-related quality of life in patients with rheumatoid arthritis——A study protocol for a multicenter, double-blind, randomized clinical trial

BACKGROUND Rheumatoid arthritis (RA) is a systemic inflammatory disease characterized by functional disability and pain. Although acupuncture is widely used, until now Western acupuncture studies on RA have not shown conclusive positive results. Acupuncture is regarded as a reflex therapy that has effects on the human autonomic nervous system. By establishing a traditional Chinese medicine (TCM) diagnosis first, the practitioner is able to choose acupoints according to the state of each individual patient. METHODS/DESIGN We are interested if acupuncture, using a classical diagnostic procedure to allocate acupoints to the patient according to the Shang Han Lun theory, can be effective in relieving pain, improving hand function and increasing health-related quality of life in RA.The authors intend to harmonize TCM diagnosis according to clinical and genetic profiles. Patients with the TCM diagnosis of a so-called Turning Point syndrome will be followed up in a randomized, prospective, double-blind, placebo-controlled, multicenter and three-armed parallel-group study with a standardized treatment in order to optimize potential therapeutic effects of acupuncture on pain, strength and muscle function of patients with RA as well as the influence on inflammation and quality of life. DISCUSSION The findings of this study will provide important clinical information about the feasibility and efficacy of acupuncture treatment for RA patients. In addition, it will explore the feasibility of further acupuncture research. TRIAL REGISTRATION NUMBER ClinicalTrials.gov Identifier NCT02553005.

Gastroprotective effect of Cymbopogon citratus infusion on acute ethanol-induced gastric lesions in rats.

ETHNOPHARMACOLOGICAL RELEVANCE Treatment of gastric ulcers with medicinal plants is quite common in traditional medicine worldwide. Cymbopogon citratus (DC) Stapf. leaves infusion has been used in folk medicine of many tropical and subtropical regions to treat gastric disturbances. The aim of this study was to assess the potential gastroprotective activity of an essential oil-free infusion from C. citratus leaves in acute gastric lesions induced by ethanol in rat. MATERIALS AND METHODS The study was performed on adult male Wistar rats (234.0±22.7g) fasted for 24h but with free access to water. The extract was given orally before (prevention) or after (treatment) intragastric administration of absolute ethanol. Effects of dose (28 or 56mg/kg of body weight) and time of contact of the extract with gastric mucosa (1 or 2h) were also assessed. Animals were sacrificed, being the stomachs removed and the lesions were assessed by macroscopic observation and histopathology. RESULTS C. citratus extract, given orally before or after ethanol, significantly (P<0.01) reduced gastric mucosal injury compared with control group (vehicle+ethanol). The effect does not appear to be dose-dependent. Results also suggested that the extract is more effective when the time of contact with gastric mucosa increases. CONCLUSIONS The results of this assay confirm the gastroprotective activity of C. citratus extract on experimental gastric lesions induced by ethanol, contributing for the pharmacological validation of its traditional use.

Fatores preditivos de complicações graves em cirurgia neonatal

OBJECTIVE To investigate the incidence and severity of early postoperative complications and to identify their risk factors in newborns undergoing surgery under general anesthesia. METHODS We conducted a retrospective analysis of data from 437 critically ill newborns undergoing surgery in a tertiary pediatric surgical center, between January 2000 and December 2010. Complications that occurred within the first 30 days after surgery were classified using the Clavien-Dindo system, for which grades III to V were considered severe. We used univariate and multivariate analysis to evaluate pre- and intraoperative variables potentially predictive of severe postoperative complications. RESULTS The incidence of at least one serious complication was 23%, with a median of one complication per patient 1:3. Altogether, there were 121 serious complications. Of these, 86 required surgical, endoscopic or radiological interventions (grade III), 25 endangered life, with uni or multi-organ failure (grade IV) and ten resulted in death (grade V). The most common complications were technical (25%), gastrointestinal (22%) and respiratory (21%). We identified four independent risk factors for severe postoperative complications: reoperation, operation for congenital diaphragmatic hernia, preterm birth less than 32 weeks of gestational age and abdominal surgery. CONCLUSION The incidence of severe postoperative complications after neonatal surgeries under general anesthesia remains high. The conditions considered independent risk factors for those can guide interventions to improve results.

Effects of the food contaminant semicarbazide on testicular morphology of juvenile Wistar rats.

O objectivo do presente trabalho foi avaliar os efeitos da semicarbazida na morfologia testicular de ratos Wistar jovens. Os animais foram tratados durante 30 dias com hidrocloreto de semicarbazida incorporado na dieta, nas concentracoes de 0, 3 e 6g/kg. Os resultados obtidos revelaram uma diminuicao estatisticamente significativa no diâmetro dos tubulos seminiferos, no diâmetro do lumen dos tubulos seminiferos e na area ocupada pelo epitelio seminifero nos animais dos grupos experimentais em comparacao com os animais do grupo controlo. Estes resultados evidenciam que a semicarbazida induz alteracoes importantes no desenvolvimento testicular e sugerem um estudo mais aprofundado sobre os mecanismos de accao, dos efeitos a longo prazo, da reversibilidade das lesoes e nas capacidades reprodutivas. Estes aspectos sao relevantes numa perspectiva de seguranca alimentar, em particular para as criancas, as quais representam um grupo de maior exposicao e susceptibilidade a semicarbazida.

Evaluation of bone turnover markers and serum minerals variations for predicting fracture healing versus non-union processes in adult sheep as a model for orthopedic research

Bone turnover markers (BTMs) have been considered as an auxiliary method of following the fracture healing process and for early prediction of impaired bone healing. A better understanding of the potential of BTMs in this application could allow for earlier interventions and improved patient care. The aim of this study with a large animal experimental model was to assess the variation of bone formation markers - namely the total alkaline phosphatase (ALP) and its bone-specific isoform (BALP), serum concentration of intact osteocalcin (OC), N-terminal propeptide type III procollagen (PIIINP) and of bone resorption markers - namely tartrate resistant acid phosphatase (TRAP) and deoxypyridinoline crosslink (DPD) during the first stages of a normal fracture healing process and of a segmental critical size defect (CSD), which progresses to a non-union process. Thirty healthy female sheep (Portuguese Churra-da-Terra-Quente breed), approximately 4-years-old, were enrolled in this study. Jugular venous blood samples were collected pre-operatively and at 1, 2, 3, 4, 6, 8, 10 and 12 post-operative weeks. The animals of the CSD group showed significant lower serum levels of BALP, OC and significant higher serum PIIINP levels at early stages of the fracture healing process, compared with animals that progressed in a normal fracture healing process. Serum BALP, OC and PIIINP levels could be useful as non-invasive auxiliary tools with other complementary methods for predicting the outcome of traumatic bone fractures.

The effectiveness of acupuncture on pain, physical function and health-related quality of life in patients with rheumatoid arthritis: a systematic review protocol

REVIEW QUESTION/OBJECTIVE The objective of this review is to identify and synthesize the best available evidence on the effectiveness of acupuncture on pain, physical function and health-related quality of life in patients with rheumatoid arthritis.More specifically, the review questions are:• Is acupuncture effective in improving health-related quality of life in patients with RA?• Is acupuncture effective in relieving pain in patients with RA?• Is acupuncture effective in improving physical function in patients with RA?