A comparison of post-thaw results between embryos arising from intracytoplasmic sperm injection using surgically retrieved or ejaculated spermatozoa

Objective: To study the effect of freeze-thaw on embryos derived from intracytoplasmic sperm injection (ICSI) using surgically retrieved and ejaculated spermatozoa. Design: Retrospective study. Setting: Private IVF center. Patient(s): Three hundred eighty-three patients undergoing frozen-thawed ET cycles. Intervention(s): Testicular sperm aspiration (TESA) or percutaneous epididymal sperm aspiration (PESA) were the sperm surgical retrieval methods used for ICSI. Embryos resulting from ICSI using Surgically retrieved and ejaculated spermatozoa were frozen, thawed, and transferred. Main Outcome Measure(s): Post-thaw survival, implantation, and pregnancy rates. Result(s): No differences were found between the ejaculated sperm and TESA/PESA groups in terms of post-thaw survival rate (68.4% vs. 66.1%, respectively), pregnancy rate (20.1% vs. 16.1%), and implantation rate (10.6% vs. 12.7%). Similar results were found for those variables when comparing TESA and PESA groups. Conclusion(s): Cleavage embryos arising from ICSI cycles using testicular and epididymal spermatozoa can be frozen with survival, pregnancy,and implantation rates comparable to those obtained with ejaculated spermatozoa. (Fertil Steril (R) 2009;91:727-32. (C) 2009 by American Society for Reproductive Medicine.)

Accuracy of working length determination using 3 electronic apex locators and direct digital radiography

Objectives. The objectives of this study were to assess the accuracy of working length determination using 3 electronic apex locators and direct digital radiography and to compare the results with those obtained using the visual method (control measurement). Study design. Twenty extracted human maxillary premolars were selected: 17 two-rooted and 3 single-rooted (total of 37 canals). Working length was measured using electronic apex locators Elements Diagnostic, Root ZX, and Just II. Subsequently, teeth were positioned in the alveolar bone of a dry skull and submitted to direct digital radiography. A variation of +/- 1 mm was considered as acceptable. Results were analyzed using the Wilcoxon and the chi(2) tests. Results. Results presented an accuracy of 94.6% for Elements Diagnostic, 91.9% for Root ZX, 73.0% for Just II, and 64.9% for direct digital radiography when considering the margin of +/- 1 mm in relation to the control measurement. Comparisons with the actual control measurements resulted in accuracy results of 13.51%, 13.51%, 10.10%, and 2.70%, respectively. Conclusions. Root ZX and Elements Diagnostic are more accurate in determining working length when compared with Just II and Schick direct digital radiography. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2011;111:e44-e49)

Determination of pulp vitality in vivo with pulse oximetry

Aim To evaluate the use of pulse oximetry as a test for pulp vitality, by comparing in the same patient, the levels of oxygen saturation of the index finger and of the maxillary central incisor and canine teeth without clinically detectable pulp inflammation. Methodology Seventeen male and female patients aged between 26 and 38 years participated and a total of 32 maxillary central incisor and 32 canine teeth were analysed. Selection criteria required the teeth to have healthy crowns, or with restorations no more than 2 mm in diameter and no clinical and radiographical signs or symptoms of pulp or periapical inflammatory changes. The negative control group consisted of 10 root filled teeth. Measurements were first taken from the index finger of patients. Their teeth were then subjected to a thermal test with refrigerant gas and then to a vitality test with pulse oximetry. Data were analysed by Pearson`s and paired t-tests. Results There were no significant statistical correlations between blood oxygen levels in the index finger and in the teeth of the patient (P > 0.05). There was a statistically significant difference in the oxygen levels between the two tooth groups studied and the index finger (P <= 0.002). Mean oxygen values in the index finger of patients were 95% (SD = 1.6), oxygen values in the maxillary central incisor were 91.29% (SD = 2.61) and mean oxygen values in maxillary canine were 90.69% (SD = 2.71). Conclusion The method determined consistently the level of blood oxygen saturation of the pulp in maxillary central incisor and canine teeth and can therefore be used for pulp vitality testing. Further studies are required to assess the effectiveness and validity of pulse oximetry in determining pulp vitality in traumatized teeth.

Gingival and bone necrosis caused by accidental sodium hypochlorite injection instead of anaesthetic solution

Aim To report clinical complications (pain, necrotic gingival tissue and bone sequestration) resulting from accidental injection of sodium hypochlorite. Summary Root canal treatment is a routine clinical procedure with few reported complications. Sodium hypochlorite (NaOCl) is commonly used as an irrigant during the procedure because of its tissue-dissolving, antibacterial and lubricating properties. This paper presents a case in which accidental injection of sodium hypochlorite into the lingual gingiva of a female patient caused gingival and bone necrosis. Surgical intervention was required. Key learning points Sodium hypochlorite is dangerous if injected into the tissues. The presentation of sodium hypochlorite in glass, anaesthetic type cartridges is potentially dangerous, and should be condemned. All healthcare workers should check carefully the contents of any syringe before injecting into patients.

An ex vivo comparison of root canal length determination by three electronic apex locators at positions short of the apical foramen

Objective. The aim of this study was to evaluate the precision of working length determination of 3 electronic apex locators (EALs): Root ZX, RomiApex D-30, and Ipex at 0.0 mm, at the apical foramen (AF), and at 1.0 mm short of the AF. Methodology. Thirty-eight mandibular premolars had their real lengths previously determined. Electronic measurements were determined at 1.0 mm, followed by measurements at 0.0 mm, performed in triplicate. Results. Precision of devices at 1.0 mm and 0.0 mm were: 94.7% and 97.4%, respectively (Root ZX); 78.9% and 97.4% (RomiApex D-30); and 76.3% and 97.4% (Ipex). Although no statistical differences were observed between the EALs at 0.0, at 1.0 mm Root ZX performed significantly better than the others. Conclusion. The EALs had acceptable precision when measuring the working length at the AF. However, when used at levels short of the AF, only Root ZX did not suffer a significant negative effect on precision. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2010;110:e57-e61)

Influence of Preflaring on the Accuracy of Length Determination With Four Electronic Apex Locators

Introduction: The aim of this study was to compare the influence of preflaring on the accuracy of 4 electronic apex locators (EALs): Root ZX, Elements Diagnostic Unit and Apex Locator, Mini Apex Locator, and Apex DSP. Methods: Forty extracted teeth were preflared by using S1 and SX ProTaper instruments. The working length was established by reducing 1 mm from the total length (TL). The ability of the EALs to detect precise (-1 mm from TL) and acceptable (-1+/-0.5 mm from TL) measurements in unflared and preflared canals was determined. Results: The precise and acceptable (P/A) readings in unflared canals for Root ZX, Elements Diagnostic Unit and Apex Locator, Mini Apex and Apex DSP were 50%/97.5%, 47.5%/95%, 50%/97.5%, and 45%/67.5%, respectively. For preflared canals, the readings were 75%/97.5%, 55%/95%, 75%/97.5%, and 60%/87.5%, respectively. For precise criteria, the preflared procedure increased the percentage of accurate electronic readings for the Root ZX and the Mini Apex Locator (P < .05). For acceptable criteria, no differences were found among Root ZX, Elements Diagnostic Unit and Apex Locator, and Mini Apex Locator (P > .05). Fisher test indicated the lower accuracy for Apex DSP (P < .05). Conclusions: The Root ZX and the Mini Apex Locator devices increased significantly the precision to determine the real working length after the preflaring procedure. All the EALs showed an acceptable determination of the working length between the ranges of+/-0.5mm except for the Apex DSP device, which had the lowest accuracy. (J Endod 2009;35:1300-1302)

Intralesional injection of triamcinolone hexacetonide as an alternative treatment for central giant-cell granuloma in 21 cases

Central giant-cell granulomas are benign, but occasionally aggressive, lesions that traditionally have been treated surgically. 21 cases of central giant-cell granuloma of the jaw were treated with intralesional injection of corticosteroids. The treatment protocol adopted was intralesional injection of 20 mg/ml triamcinolone hexacetonide diluted in an anaesthetic solution of 2% lidocaine/epinephrine 1:200,000 in the proportion 1:1; 1.0 ml of the solution was infiltrated for every 1 cm(3) of radiolucid area of the lesion, totalling 6 biweekly applications. Ten patients had aggressive lesions and 11 nonaggressive. Two patients showed a negative response to the treatment and underwent surgical resection, 4 showed a moderate response and 15 a good response. 8 of the 19 who had a moderate-to-good response to the drug treatment underwent osteoplasty to reestablish facial aesthetics. In these cases, only mature or dysplastic bone was observed, with the presence or absence of rare giant multinucleated cells. The advantages of this therapy are its less-invasive nature, the probable lower cost to the patient, lower risk and the ability to treat the lesion surgically in the future, if necessary.

Central NO-cGMP pathway in thermoregulation and survival rate during polymicrobial sepsis

Sepsis induces production of inflammatory mediators such as nitric oxide (NO) and causes physiological alterations, including changes in body temperature (T(b)). We evaluated the involvement of the central NO cGMP pathway in thermoregulation during sepsis induced by cecal ligation and puncture (CLP), and analyzed its effect on survival rate. Male Wistar rats with a T(b) probe inserted in their abdomen were intracerebroventricularly injected with 1 mu L N(G)-nitro-L-arginine methyl ester (L-NAME, 250 mu g), a nonselective NO synthase (NOS) inhibitor; or aminoguanidine (250 mu g), an inducible NOS inhibitor; or 1H-[1,2,4]oxadiazolo[4,3,-a]quinoxalin-1-one (ODQ, 0.25 mu g), a guanylate cyclase inhibitor. Thirty minutes after injection, sepsis was induced by cecal ligation and puncture (CLP), or the rats were sham operated. The animals were divided into 2 groups for determination of T(b) for 24 h and assessment of survival during 3 days. The drop in T(b) seen in the CLP group was attenuated by pretreatment with the NOS inhibitors (p < 0.05) and blocked with ODQ. CLP rats pretreated with either of the inhibitors showed higher survival rates than vehicle injected groups (p < 0.05), and were even higher in the ODQ pretreated group. Our results showed that the effect of NOS inhibition on the hypothermic response to CLP is consistent with the role of nitrergic pathways in thermoregulation.

Evaluation of estrogen neuroprotective effect on nigrostriatal dopaminergic neurons following 6-hydroxydopamine injection into the substantia nigra pars compacta or the medial forebrain bundle

Studies involving estrogen treatment of ovariectomized rats or mice have attributed to this hormone a neuroprotective effect on the substantia nigra pars compacta (SNpc) neurons. We investigated the effect of estradiol replacement in ovariectomized rats on the survival of dopaminergic mesencephalic cell and the integrity of their projections to the striatum after microinjections of 1 mu g of 6-hydroxydopamine (6-OHDA) into the right SNpc or medial forebrain bundle (MFB). Estradiol replacement did not prevent the reduction either in the striatal concentrations of DA and metabolites or in the number of nigrostriatal dopaminergic neurons following lesion with 1 mu g of 6-OHDA into the SNpc. Nevertheless, estradiol treatment reduced the decrease in striatal DA following injection of 1 mu g of 6-OHDA into the MFB. Results suggest therefore that estrogen protect nigrostriatal dopaminergic neurons against a 6-OHDA injury to the MFB but not the SNpc. This may be due to the distinct degree of lesions promoted in these different rat models of Parkinson`s disease.

Determination of para-Chloroaniline and Reactive Oxygen Species in Chlorhexidine and Chlorhexidine Associated with Calcium Hydroxide

The aim of this study was to determine whether para-chloroaniline (PCA) and/or reactive oxygen species (ROS) are generated by chlorhexidine (CHX) alone or after CHX is mixed with calcium hydroxide at different time points. Mass spectrometry was performed to detect PCA in samples of 0.2% CHX and Ca(OH)2 mixed with 0.2% CHX. High-performance liquid chromatography was used to confirm the presence of CHX in the mixture with Ca(OH)2. The samples were analyzed immediately after mixing and after 7 and 14 days. During the intervals of the experiment, the samples were maintained at 36.5 degrees C and 95% relative humidity. PCA was detected in the 0.2% CHX solution after 14 days. The mixture of CHX with Ca(CH)2 liberated ROS at all time points, but no traces of CHX were present in the mixture as a result of immediate degradation of the CHX. (J Endod 2008;34:1508-1514)

Solid-state NMR structure determination of melittin in a lipid environment

Solid-state C-13 NMR spectroscopy was used to investigate the three-dimensional structure of melittin as lyophilized powder and in ditetradecylphosphatidylcholine (DTPC) membranes. The distance between specifically labeled carbons in analogs [1-C-13]Gly3-[2-C-13]Ala4, [1-C-13]Gly3-[2-C-13]Leu6, [1-C-13]Leu13-[2-C-13]Ala15, [2-C-13]Leu13-[1-C-13]Ala15, and [1-C-13]Leu13-[2-C-13]Leu16 was measured by rotational resonance. As expected, the internuclear distances measured in [1-C-13]Gly3-[2-C-13]Ala4 and [1-C-13]Gly3-[2-C-13]Leu6 were consistent with alpha -helical structure in the N-terminus irrespective of environment. The Internuclear distances measured in [1-C-13]Leu13-[2-C-13]Ala15, [2-C-13]Leu13-[1-C-13]Ala15, and [1-C-13]Leu13-[2-C-13]Leu16 revealed, via molecular modeling, some dependence upon environment for conformation in the region of the bend in helical structure induced by Pro14. A slightly larger interhelical angle between the N- and C-terminal helices was indicated for peptide in dry or hydrated gel state DTPC (139 degrees -145 degrees) than in lyophilized powder (121 degrees -139 degrees) or crystals (129 degrees). The angle, however, is not as great as deduced for melittin in aligned bilayers of DTPC in the liquid-crystalline state (similar to 160 degrees) (R. Smith, F. Separovic, T. J. Milne, A. Whittaker, F. M. Bennett, B. A. Cornell, and A. Makriyannis, 1994, J. Mol, Biol 241:456-466). The study illustrates the utility of rotational resonance in determining local structure within peptide-lipid complexes.

Determination of the intramolecular disulfide bond arrangement and biochemical identification of the glycosylation sites of the nonstructural protein NS1 of Murray Valley encephalitis virus

The 12 cysteine residues in the flavivirus NS1 protein are strictly conserved, suggesting that they form disulfide bonds that are critical for folding the protein into a functional structure. In this study, we examined the intramolecular disulfide bond arrangement of NS1 of Murray Valley encephalitis virus and elucidated three of the six cysteine-pairing arrangements. Disulfide linkages were identified by separating tryptic-digested NS1 by reverse-phase high pressure liquid chromatography and analysing the resulting peptide peaks by protein sequencing, amino acid analysis and/or electrospray mass spectrometry. The pairing arrangements between the six amino-terminal cysteines were identified as follows: Cys(4)-Cys(15), Cys(55)-Cys(143) and Cys(179)-Cys(223). Although the pairing arrangements between the six carboxyterminal cysteines were not determined, we were able to eliminate several cysteine-pairing combinations. Furthermore, we demonstrated that all three putative N-linked glycosylation sites of NS1 are utilized and that the Asn(207) glycosylation site contains a mannose-rich glycan.