Aneural culture of rat myoblasts for myocardial transplant

Due to the peculiar characteristics of skeletal muscle, myoblast transplants have emerged as a therapy for cardiomyopathy, particularly after myocardial infarction. The objectives of this study were to define the mean time of cultivation necessary to obtain a cellular concentration of 10(6) to expand the mass for transplant, and to identify the proliferation phase of myoblasts. Ten myoblast cultures were performed using newborn Wistar rats. The isolation method used enzymatic dissociation in culture medium (HAM-F12 and 199) supplement with basic-fibroblast growth factor (b-FGF) and insulin growth factor (IGF-I). The mean cultivation time to obtain the desired concentration of 10(6) was 7 days, with expansion of up to 10(8)/g. When b-FGF was used, the cellular yield was approximately 10(7), with IGF-I the cellular yield was approximately 10(8), independent of the medium. We concluded that IGF-I is the better option for mass cellular expansion of myoblasts for application in myocardial transplants.     

Snake venom phospholipase A2s (Asp49 and Lys49) induce mechanical allodynia upon peri-sciatic administration: involvement of spinal cord glia, proinflammatory cytokines and nitric oxide

Snakebites constitute a serious public health problem in Central and South America, where species of the lancehead pit vipers (genus Bothrops) cause the majority of accidents. Bothrops envenomations are very painful, and this effect is not neutralized by antivenom treatment. Two variants of secretory phospholipases A2 (sPLA2), corresponding to Asp49 and Lys49 PLA2s, have been isolated from Bothrops asper venom. These sPLA2s induce hyperalgesia in rats following subcutaneous injection. However, venom in natural Bothrops bites is frequently delivered intramuscularly, thereby potentially reaching peripheral nerve bundles. Thus, the present series of experiments tested whether these sPLA2s could exert pain-enhancing effects following administration around healthy sciatic nerve. Both were found to produce mechanical allodynia ipsilateral to the injection site; no thermal hyperalgesia was observed. As no prior study has examined potential spinal mechanisms underlying sPLA2 actions, a series of anatomical and pharmacological studies were performed. These demonstrated that both sPLA2s produce activation of dorsal horn astrocytes and microglia that is more prominent ipsilateral to the site of injection. As proinflammatory cytokines and nitric oxide have each been previously implicated in spinally mediated pain facilitation, the effect of pharmacological blockade of these substances was tested. The results demonstrate that mechanical allodynia induced by both sPLA2s is blocked by interleukin-1 receptor antagonist, anti-rat interleukin-6 neutralizing antibody, the anti-inflammatory cytokine interleukin-10, and a nitric oxide synthesis inhibitor (L-NAME). As a variety of immune cells also produce and release sPLA2s during inflammatory states, the data may have general implications for the understanding of inflammatory pain.     

Snake venom components enhance pain upon subcutaneous injection: an initial examination of spinal cord mediators

Snakebites are a relevant public health problem in Central and South America. Snake bite envenomations cause intense pain, not relieved by anti-venom. The fangs of many species are short, causing subcutaneous injection. Fangs of larger species inflict subcutaneous or intramuscular envenomation. To understand pain induced by subcutaneous venom, this study examined spinal mechanisms involved in pain-enhancing effects of subcutaneous Lys49 and Asp49 secretory phospholipase-A(2) (sPLA2), two components of Bothrops asper snake venom showing highly different enzymatic activities. Unilateral intraplantar sPLA2-Lys49 (catalytically inactive) or sPLA2-Asp49 (catalytically active) into rat hindpaws each induced mechanical hyperalgesia (Randall-Selitto test), whereas only catalytically active sPLA2-Asp49 caused mechanical allodynia (von Frey test). Effects induced by both sPLA2s were inhibited by intrathecal fluorocitrate, a reversible glial metabolic inhibitor. In support, immunohistochemical analysis revealed activation of dorsal horn astrocytes and microglia after intraplantar injection of either sPLA2. Spinal proinflammatory cytokines, nitric oxide, and prostanoids each appear to be involved in the pain-enhancing effects of these sPLA2s. Blockade of interleukin-1 (IL1) inhibited hyperalgesia induced by both sPLA2s, while leaving allodynia unaffected. Blockade of tumor necrosis factor reduced responses to sPLA2-Asp49. An inhibitor of neuronal nitric oxide synthase, 7-nitroindazole (7-NI), inhibited hyperalgesia induced by both sPLA2s, without interfering with allodynia induced by sPLA2-Asp49. On the other hand, L-N(6)-(1-iminoethyl)lysine (L-NI), an inhibitor of the inducible nitric oxide synthase, did not alter any sPLA2-induced effect. Lastly, celecoxib, an inhibitor of cyclooxygenase-2, attenuated sPLA2 actions. These data provide the first evidence of spinal mediators involved in pain facilitation induced by subcutaneous venoms.     

Reference range for cervical length in twin pregnancies

BACKGROUND: Twin pregnancies are at increased risk of preterm delivery, and cervical length measurement by transvaginal ultrasound examination has been shown to be an effective method of screening for this complication. The aim of the present study is to establish a reference range for cervical length measurement throughout gestation in twin pregnancies. METHODS: Prospective study involving 144 nonselected twin pregnancies that delivered after 34 weeks and underwent ultrasound examination every 4 weeks for transvaginal measurement of cervical length. Mean cervical length measurements, 95% confidence intervals for the estimate and 95% prediction intervals were established throughout gestation by linear regression analysis. RESULTS: There was a significant correlation between cervical length measurements and gestational age [cervical length (mm) = - 0.758 * gestation age (weeks) + 56.69, r=- 0.43, p < 0.0001], and mean cervical length shortened by approximately 0.8 mm per week, from 47 mm at 13 weeks to 32 mm at 32 weeks. The corresponding lower limits for the 95% prediction intervals were 29 and 15 mm, respectively. CONCLUSION: Cervical length shortens gradually throughout gestation in twin pregnancies.     

Effect of a lactose-containing sweetener on root dentine demineralization in situ

Since lactose has been used as an excipient in powdered sweeteners, we evaluated its effect on dentine demineralization, in an in situ crossover study. Twelve adult volunteers wore palatal appliances containing 4 blocks of bovine root dentine, which were submitted 4 times a day to: distilled water (negative control), aqueous solution of powdered sweetener (Zero Cal); experimental group), 1.5% lactose solution (active group), 1.5% sucrose solution (positive control). After 15 days, microhardness was determined on dentine surface (SMH) and on the sectioned blocks (CSMH). All groups presented statistical differences in SMH before and after the treatments; water increased, while sweetener, lactose and sucrose decreased SMH. By CSMH, the only significant difference was observed after sucrose treatment. The data suggest that sweeteners containing lactose as a bulking agent may be cariogenic for root dentine.     

Effect of cooling procedure on final denture base adaptation

Well-fitted dentures prevent hyperplasic lesions, provide chewing efficiency and promote patient's comfort. Several factors may affect final adaptation of dentures, as the type of the acrylic resin, the flask cooling procedure and the water uptake. This investigation evaluated the effect of water storage and two different cooling procedures [bench cooling (BC) for 2 h; running water (RW) at 20 degrees C for 45 min] on the final adaptation of denture bases. A heat-cured acrylic resin (CL, Clássico, Clássico Artigos Odontológicos) and two microwave-cured acrylic resins [Acron MC, (AC) GC Dent. Ind. Corp.; Onda Cryl (OC), Clássico Artigos Odontológicos] were used to make the bases. Adaptation was assessed by measuring the weight of an intervening layer of silicone impression material between the base and the master die. Data was submitted to ANOVA and Tukey's test (0.05). The following means were found: (BC) CL=0.72 +/- 0.03 a; AC=0.70 +/- 0.03 b; OC=0.76 +/- 0.04 c//(RW) CL= 1.00 +/- 0.11 a; AC=1.00 +/- 0.12 a; OC=0.95 +/- 0.10 a. Different labels join groups that are not statistically different (P > 0.05). Comparisons are made among groups submitted to the same cooling procedure (BC or RW). The conclusions are: interaction of type of material and cooling procedure had a statistically significant effect on the final adaptation of the denture bases (P < 0.05); water storage was not detected as a source of variance (P > 0.05) on the final adaptation.     

Microwave-cured acrylic resins and silicone-gypsum moulding technique

This study evaluated the residual monomer (RM), Knoop hardness (KHN) and transverse strength (TS) of two microwave‐cured acrylic resins (Acron MC^® (A), GC Dent. Ind. Corp., Tokyo, Japan and Onda Cryl^® (O), AO Classico Ltda, São Paulo, Brazil) when processed with an all‐type III gypsum moulding technique (G) or a silicone–gypsum moulding technique (S). One hundred and forty four specimens were fabricated and equally distributed into four groups (AG, AS, OG and OS). The TS tests were conducted after 48 h of water storage, and KHN values were obtained after 24, 48, 72 h and 30 days. The RM was determined every 24 or 48 h over a period of 288 h. The acrylic resins were prepared and processed according to the manufacturer’s directions. Both AS and OS showed the highest means (P < 0·05) for RM (μg cm^–2) only after 24 h (AS = 56·84 ± 27·39/AG = 7·51 ± 5·75/OS = 3·59 ± 1·60/OG = 1·02 ± 0·3), 48 h (AS = 28·99 ± 9·35/AG = 2·65 ± 2·17/OS = 2·37 ± 0·84/OG = 0·68 ± 0·49) and 72 h (AS = 15·98 ± 9·01/AG = 1·40 ± 0·57/OS = 1·87 ± 0·52/OG = 0·75 ± 0·44). Both AS and OS showed the highest means (P < 0·02) for KH after 24 h (AS = 18·69 ± 2·3/AG = 17·79 ± 0·7/OS = 18·41 ± 1·0/OG = 16·04 ± 0·6). After 48 h the mean values for OS and OG differed significantly (P < 0·03) (OS = 18·67 ± 0·8/OG = 16·75 ± 0·8). No differences of KHN were found among the groups during the storage periods. The TS values for A and O were not affected by either G or S (P > 0·05). Silicone–gypsum mould technique affected the RM and KHN of the resins in the first 2 and 5 days of analysis, respectively. The type of mould did not affect TS, and the acrylic resins differed from each other for all properties regardless of the type of mould.     

Rationale and design of the ViCTORY (Validation of an Intracycle CT Motion CORrection Algorithm for Diagnostic AccuracY) trial

BackgroundCoronary CT angiography (CTA) has emerged as an effective noninvasive method for direct visualization of the coronary arteries, with high diagnostic performance compared with invasive coronary angiography (ICA). However, coronary CTA is prone to artifacts, including coronary motion, which may reduce its diagnostic performance. Intracycle motion compensation algorithms (MCAs) from a combination of software and hardware techniques now allow for correction of coronary motion, but the diagnostic performance of MCAs compared with traditional coronary CTA reconstruction methods remains unexplored.MethodsViCTORY (Validation of an Intracycle CT Motion CORrection Algorithm for Diagnostic AccuracY) is a prospective international multicenter trial of 218 patients which is designed to evaluate the performance of MCAs for the diagnosis of anatomically obstructive coronary artery disease (CAD) compared with an ICA reference standard, on a per-patient, per-vessel, and per-segment basis. Patients enrolled into ViCTORY will undergo investigational coronary CTA and clinically indicated ICA and will not receive heart rate-lowering medications before coronary CTA. Coronary CTA images will be reconstructed by conventional standard methods as well as by MCAs. Blinded core laboratory interpretation will be performed for coronary CTA and ICA in an intent-to-diagnose fashion.ResultsThe primary end point of ViCTORY is the per-patient diagnostic accuracy of MCAs for the diagnosis of anatomically obstructive CAD compared with ICA. Secondary end points will include other per-patient, per-vessel, and per-segment diagnostic performance characteristics, including accuracy, sensitivity, specificity, positive predictive value, and negative predictive value. Other key secondary end points will include diagnostic interpretability, image quality, the upper heart rate threshold of utility of MCAs, and the additive value of MCAs to traditionally reconstructed coronary CTA.ConclusionViCTORY will determine whether MCAs improve the diagnosis of obstructive CAD in patients undergoing coronary CTA who are not receiving heart rate-lowering medications.