Evaluation of three substitutes for Percoll in sperm isolation by density gradient centrifugation

Silane-coated silica particle solutions (ISolate(TM) and PureSperm)TM)) and iodixanol (OptiPrep(TM)) were compared to polyvinylpyrrolidone (PVP)-coated silica particles (Percoll(TM)) in their efficacy to recover spermatozoa by gradient centrifugation for use in assisted reproductive procedures. Efficacy was assessed in terms of percentages of sperm recovery, sperm vitality and motility, normal sperm morphology and normal sperm chromatin condensation. No significant difference was found in the recovery of spermatozoa for men with both normal sperm counts and oligozoospermia, between PVP-coated and silane-coated particle solutions. Iodixanol had significantly lower sperm recovery compared to the other products. Sperm vitality, progressive motility, normal morphology and normal chromatin condensation did not differ significantly between any of the sperm isolation products.      

Protective cytotoxic T lymphocyte responses against paramyxoviruses induced by epitope-based DNA vaccines: involvement of IFN-gamma

Plasmid DNA vectors have been constructed with minigenes encoding a single cytotoxic T lymphocyte (CTL) epitope from either the M2 protein of respiratory syncytial virus (RSV) or from the nucleoprotein of measles virus (MV) with or without a signal sequence (also called secretory or leader sequence). Following intradermal immunization, plasmids in which the CTL epitopes were expressed in-frame with the signal sequence were more effective at inducing peptide- and virus- specific CTL responses than plasmids expressing CTL epitopes without the signal sequence. This immunization resulted in protection against MV-induced encephalitis and a significant reduction in viral load following RSV challenge. The reduction of viral load following RSV challenge was abrogated by prior injection with anti-IFN-gamma antibodies. These results highlight the ability of epitope-based DNA immunization to induce protective immune responses to well-defined epitopes and indicate the potential of this approach for the development of vaccines against infectious diseases.      

CBCT-based predictability of attachment of the neurovascular bundle to the proximal segment of the mandible during sagittal split osteotomy

Injury to the inferior alveolar nerve (IAN) during surgery is an important complication of bilateral sagittal split osteotomy. With cone beam computed tomography, the course of the nerve and its relationship to the surrounding structures can be assessed in three dimensions. This study aims to determine whether tomography can predict attachment of the neurovascular bundle to the proximal segment of the mandible during sagittal split osteotomy (SSO). Bilateral linear measurements were taken on cross-sectional tomography images. During osteotomy, it was noted for each patient whether the neurovascular bundle was attached to the proximal segment during the split. If attached, a bone-cutting instrument or a blunt instrument was needed to free the nerve. The nerve was attached at more than one-third of operation sites (170 sites). Of these, over 65% of attached nerves (108 sites) required a bone-cutting intervention to free them from the mandible. After correcting for confounding factors, the linear distances from the buccal cortical margin of the IAN canal to the inner and outer buccal cortical margins of the mandible were important predictors of whether the IAN will be attached to the proximal segment of the mandible during SSO.     

Anicteric hepatoxicity: a potential health risk of occupational exposures in Nigerian petroleum oil refining and distribution industry

Background

Literature abounds linking one’s job to certain unpalatable health outcomes. Since exposures to hazardous conditions in industrial environments often results in sundry health effects among workers, we embarked on this study to investigate the hepatic health effects of occupational activities in the petroleum refining and distribution industry.

Method

Biochemical markers of liver functions were assayed in plasma, using Reflotron dry chemistry spectrophotometric system. The study was conducted on randomly selected workers of Port Harcourt Refining Company (PHRC) and Pipelines and Petroleum Product Marketing Company (PPMC) both in Alesa-Eleme near Port Harcourt, Nigeria, as well as non-oil work civil servants serving as control subjects.

Result and conclusion

Results showed that, bilirubin ranged 0.3-1.6 mg/dl with a mean of 0.66±0.20mg/dl among the oil workers as against 0.5-1.00mg/dl with a mean of 0.58±0.13mg/dl in non-oil workers, Alkaline phosphatase ranged 50.00-296.00u/l (mean: 126.21±39.49u/l) in oil workers as against 40.20-111u/l (mean: 66.83±18.54u/l) for non-oil workers, Aspartic transaminases (AST) ranged 5.80-140.20u/l (mean: 21.81±11.49u/l) in oil workers against 18.00-44.00u/l (mean: 26.89±6.99u/l) for non-oil workers, while Alanine transaminases (ALT) ranged 4.90-86.00u/l (mean: 22.14±11.28u/l) in oil workers as against 10.00-86.60u/l (mean: 22.30±10.22u/l) for the non-oil workers. A close study of the results revealed that although the mean values for all the studied parameters were still within the parametric reference ranges, however, relative to the referents, there were significant increases (P<0.05) in plasma bilirubin (though anicteric) and alkaline phosphatase that was not matched with a corresponding increase in the plasma transaminases, suggesting a possibility that toxic anicteric hepatoxicity is part of the potential health effects of sundry exposures in the Nigeria petroleum oil refining and distribution industry. Gender differentiation data showed that though the mean values for the parameters were higher in males than females, the increases were not significant in most cases (P>0.05), whereas data for age and exposure period classifications revealed that irrespective of the age of the worker, the effects are likely to start after the first five years, manifesting fully after the first decade of occupational exposures. Thus, an update of industrial/occupational health measures is necessary for a safer and healthier work environment.     

Agreement between quantitative and qualitative sensory testing of changes in oro‐facial somatosensory sensitivity

Qualitative somatosensory testing (QualST) is a simple chairside test. It can be used to roughly assess the presence or absence of altered somatosensory function. To use QualST clinically, it is important to assess its agreement with quantitative sensory testing (QST). The aims of this study were to assess the agreement between QST and QualST when testing the modulation of facial sensitivity by capsaicin in healthy participants and to explore the agreement between QST and QualST in assessing the intraoral sensory function in clinical atypical odontalgia (AO) patients. Eighteen healthy pain‐free adults and data from 27 AO patients were included in the study. Thirteen QST and three QualST parameters were evaluated at each site. Z‐scores were computed for healthy participants, and Loss‐Gain scores were created. The agreement observed between QST and QualST in participants with no alterations in facial sensation (placebo) was good, that is ranging from 89% to 94%. A poorer agreement was seen after capsaicin application in all test modalities with agreement ranging from 50% to 72%. The commonest misclassification observed was participants classified as normal according to QST, but hyper‐ or hyposensitive according to QualST after capsaicin application, especially for cold and pinprick. A similar trend was observed in AO patients where patients classified as normal using QST were misclassified as hypersensitive and in few patients as hyposensitive by QualST. In conclusion, the study showed that QualST may be used as a screening tool in the clinical setting, especially to show that subjects have normal sensory function.     

Analysis of the relationship between age and treatment response in migraine

In migraine, headache severity varies with age. As a consequence, the effectiveness of medication may also depend on a patient's age. The purpose of this study was to assess the combined effect of age and drug treatment on headache characteristics. Using data from clinical trials of sumatriptan in adolescents and adults, we show how the interaction between age and drug exposure can be parameterised as a covariate on a Markov model that describes the decline of headache severity over three clinically defined stages (no relief, relief and pain-free status). The model explains important clinical observations: (i) the rates at which the pain relief and pain-free status were attained were found to be inversely related to age; (ii) in placebo-treated patients, the mean transit time from 'no relief' to 'relief' is 3 h for young adolescents and increases to 6 h for patients aged ≥ 30 years; and (iii) sumatriptan reduces the transit time to 2 h, irrespective of age. These findings indicate that the therapeutic gain over placebo increases with age. Prospective studies of antimigraine drugs should take this relationship into account when extrapolating efficacy data from adults to adolescents.     

Metabolic profile changes in the testes of mice with streptozotocin-induced type 1 diabetes mellitus

Contrary to the traditional view, recent studies suggest that diabetes mellitus has an adverse influence on male reproductive function. Our aim was to determine the effect of diabetes on the testicular environment by identifying and then assessing perturbations in small molecule metabolites. Testes were obtained from control and streptozotocin-induced diabetic C57BL/6 mice, 2, 4 and 8 weeks post-treatment. Diabetic status was confirmed by glycated haemoglobin, non-fasting blood glucose, physiological condition and body weight. A novel extraction procedure was utilized to obtain protein free, low-molecular weight, water soluble extracts which were then assessed using ¹H nuclear magnetic resonance spectroscopy. Principal component analysis of the derived profiles was used to classify any variations, and specific metabolites were identified based on their spectral pattern. Characteristic metabolite profiles were identified for control and type 1 diabetic animals with the most distinctive being from mice with the largest physical deterioration and loss of body weight. Eight streptozotocin-treated animals did not develop diabetes and displayed profiles similar to controls. Diabetic mice had decreases in creatine, choline and carnitine and increases in lactate, alanine and myo-inositol. Betaine levels were found to be increased in the majority of diabetic mice but decreased in a few animals with severe loss of body weight and physical condition. The association between perturbations in a number of small molecule metabolites known to be influential in sperm function, with diabetic status and physiological condition, adds further impetus to the proposal that diabetes influences important spermatogenic pathways and mechanisms in a subtle and previously unrecognized manner.     

Male diabetes mellitus and assisted reproduction treatment outcome

The long-held view that diabetes has little effect on male reproductive function has been challenged by findings that the condition influences fertility in numerous previously undetected ways. This retrospective chart review of 3000 couples determined the incidence of couples with a male diabetic seeking assisted reproduction treatment and assessed any relationship between male diabetes and IVF/intracytoplasmic sperm injection (ICSI) outcome. Eight (2.7%) couples were found with a diabetic male partner, of which 18 couples underwent assisted reproduction treatment (five IVF, 12 ICSI, one both), with fertilization rates (IVF 68%, ICSI 62%) similar to non-diabetic patients (IVF 70%, ICSI 71%) and no difference in embryo quality. Two men had retrograde ejaculation and two were azoospermic. Other than reduced sperm motility, the remaining 14 had normal World Health Organization semen parameters. Embryo transfers produced one pregnancy (5% combined IVF/ICSI pregnancy rate/cycle) giving a lower-than-expected rate (28.8%). The pregnancy rate from seven FETs (29%) was comparable to the expected (21.3%). Compared with non-diabetics, approximately three times more couples with diabetic men sought treatment, with a larger percentage having 'unexplained' infertility. Fertilization rates and embryo quality did not differ but pregnancy rates were lower in couples with a diabetic male.     

Validation of anatomical landmarks-based registration for image-guided surgery: An in-vitro study

IntroductionPerioperative navigation is a recent addition to orthognathic surgery. This study aimed to evaluate the accuracy of anatomical landmarks-based registration.Materials and methodsEighty-five holes (1.2 mm diameter) were drilled in the surface of a plastic skull model, which was then scanned using a SkyView cone beam computed tomography scanner. DICOM files were imported into BrainLab ENT 3.0.0 to make a surgical plan. Six anatomical points were selected for registration: the infraorbital foramena, the anterior nasal spine, the crown tips of the upper canines, and the mesial contact point of the upper incisors. Each registration was performed five times by two separate observers (10 times total).ResultsThe mean target registration error (TRE) in the anterior maxillary/zygomatic region was 0.93 ± 0.31 mm (p < 0.001 compared with other anatomical regions). The only statistically significant inter-observer difference of mean TRE was at the zygomatic arch, but was not clinically relevant.ConclusionWith six anatomical landmarks used, the mean TRE was clinically acceptable in the maxillary/zygomatic region. This registration technique may be used to access occlusal changes during bimaxillary surgery, but should be used with caution in other anatomical regions of the skull because of the large TRE observed.