Fabrication and evaluation of porous coatings doped with bioactive elements on titanium surfaces.

OBJECTIVE: Although pure titanium (PT) and its alloys exhibit excellent mechanical properties, they lack biological activity as implants. The purpose of this study was to improve the biological activity of titanium implants through surface modification. MATERIALS AND METHODS: Titanium was processed into titanium discs, where the titanium discs served as anodes and stainless steel served as cathodes, and a copper- and cobalt-doped porous coating [pure titanium model (PTM)] was prepared on the surface of titanium via plasma electrolytic oxidation. The surface characteristics of the coating were evaluated using field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), and profilometry. The corrosion resistance of PTM was evaluated with an electrochemical workstation. The biocompatibility and bioactivity of coated bone marrow mesenchymal stem cells (BMSCs) were evaluated through in vitro cell experiments. RESULTS: A copper- and cobalt-doped porous coating was successfully prepared on the surface of titanium, and the doping of copper and cobalt did not change the surface topography of the coating. The porous coating increased the surface roughness of titanium and improved its resistance to corrosion. In addition, the porous coating doped with copper and cobalt promoted the adhesion and spreading of BMSCs. CONCLUSIONS: A porous coating doped with copper and cobalt was prepared on the surface of titanium through plasma electrolytic oxidation. The coating not only improved the roughness and corrosion resistance of titanium but also exhibited good biological activity.

[Early results of modified Bikini approach periacetabular osteotomy for the treatment of developmental hip dysplasia under 50 years of age].

Objective: To investigate the efficacy and safety of modified Bikini approach periacetabular osteotomy in the treatment of developmental hip dysplasia under 50 years of age. Methods: The clinical data of 39 patients with developmental hip dysplasia who underwent periacetabular osteotomy in the Department of Orthopedics, Guizhou Provincial People's Hospital from June 2016 to June 2021 were retrospectively analyzed.Among them, 20 patients (21 hips) underwent the improved Bikini approach (study group) and 19 patients (20 hips) underwent the improved Smith-Petersen approach (control group).In the study group, there were 3 males and 17 females, aged(M(IQR))27.5 (14.3) years (range:11 to 44 years).In the control group, there were 2 males and 17 females, aged 27.5 (19.3) years (range:17 to 47 years).Both groups were sutured in the same manner by the same physician.Incision length, operation time, intraoperative blood loss and complications were recorded.X-ray images, anterior central marginal angle (ACE), lateral central marginal Angle (LCE) and acetabulum tilt angle (Tonnis AI) were measured before and after the operation.The coverage rate of acetabulum to femoral head (AHI) was measured and calculated, and the healing time was observed.Harris Hip score, International Hip score (IHOT)-12 and visual analogue scale (VAS) were recorded before and after surgery.Vancouver Scar Scale (VSS) score and patient and observer scar assessment scale (POSAS) score were recorded 12 months after surgery.The independent sample t test,Wilcoxon rank sum test, χ2 test or Fisher exact test were used to compare the clinical efficacy between the two groups. Results: All patients successfully completed the operation.There was no significant difference in operation time and intraoperative blood loss between the two groups (all P>0.05).The incision length of the study group was smaller than that of the control group, and the difference was statistically significant (10.5(5.0)cm vs.15.0(3.0),W=309.000,P=0.007).Patients were followed up for (19.1±11.1) months (range:12 to 60 months).Femoral nerve stretching injury occurred in 2 cases and sciatic branch fracture occurred in 1 case in the study group, all of which recovered to normal at 3 months follow-up, while no corresponding injury occurred in the control group.Lateral femoral cutaneous nerve injury occurred in 3 cases in the study group and 2 cases in the control group.Delayed wound healing occurred in 1 case in each of the two groups, and both healed after re-operation debridement and suture.Pubic branch nonunion occurred in 4 patients in the study group and 5 patients in the control group.There were no serious complications such as sciatic nerve and femoral blood vessel injury between the 2 groups, and there was no statistical significance in the incidence of complications between the 2 groups (52.4%(11/21)vs.40.0%(8/20),χ2=0.631,P=0.427).The clinical healing time of the patient was (4.5±1.3) months after surgery (range:3.0 to 8.0 months).There were no significant differences in ACE, LCE, Tonnis AI and AHI between the 2 groups (all P>0.05).At the last follow-up, there were no significant differences in VAS,Harris hip score and IHOT-12 score between the two groups (all P>0.05).The incision scars in the study group were smaller than those in the control group, and the differences in VSS and POSAS were statistically significant (all P<0.05). Conclusion: Compared with the improved Smith-Petersen approach, the improved Bikini approach has the same early clinical efficacy in the treatment of patients with developmental hip dysplasia under the age of 50, and has the advantages of smaller postoperative incision scars, more hidden and beautiful incision, and no serious complications, which is worthy of further study and promotion.

Vagus nerve stimulation promotes gastric emptying by increasing pyloric opening measured with magnetic resonance imaging.

BACKGROUND: Vagus nerve stimulation (VNS) is an emerging electroceutical therapy for remedying gastric disorders that are poorly managed by pharmacological treatments and/or dietary changes. Such therapy seems promising as the vagovagal neurocircuitry modulates the enteric nervous system to influence gastric functions. METHODS: Here, the modulatory effects of left cervical VNS on gastric emptying in rats were quantified using a (i) feeding protocol in which the animal voluntarily consumed a postfast, gadolinium-labeled meal and (ii) a non-invasive imaging method to measure antral motility, pyloric activity and gastric emptying based on contrast-enhanced magnetic resonance imaging (MRI) and computer-assisted image processing pipelines. KEY RESULTS: Vagus nerve stimulation significantly accelerated gastric emptying (sham vs VNS: 29.1% ± 1.5% vs 40.7% ± 3.9% of meal emptied per 4 hours), caused a greater relaxation of the pyloric sphincter (sham vs VNS: 1.5 ± 0.1 vs 2.6 ± 0.4 mm2 cross-sectional area of lumen), and increased antral contraction amplitude (sham vs VNS: 23.3% ± 3.0% vs 32.5% ± 3.0% occlusion), peristaltic velocity (sham vs VNS: 0.50 ± 0.02 vs 0.67 ± 0.03 mm s-1 ), but not its contraction frequency (sham vs VNS: 6.1 ± 0.2 vs 6.4 ± 0.2 contractions per minute, P = .22). The degree to which VNS relaxed the pylorus was positively correlated with gastric emptying rate (r = .5887, P < .001). CONCLUSIONS & INFERENCES: The MRI protocol employed in this study is expected to enable advanced preclinical studies to understand stomach pathophysiology and its therapeutics. Results from this study suggest an electroceutical treatment approach for gastric emptying disorders using cervical VNS to control the degree of pyloric sphincter relaxation.

Phase I-II clinical trial design: a state-of-the-art paradigm for dose finding.

Background: Conventional phase I algorithms for finding a phase-2 recommended dose (P2RD) based on toxicity alone is problematic because the maximum tolerated dose (MTD) is not necessarily the optimal dose with the most desirable risk-benefit trade-off. Moreover, the increasingly common practice of treating an expansion cohort at a chosen MTD has undesirable consequences that may not be obvious. Patients and methods: We review the phase I-II paradigm and the EffTox design, which utilizes both efficacy and toxicity to choose optimal doses for successive patient cohorts and find the optimal P2RD. We conduct a computer simulation study to compare the performance of the EffTox design with the traditional 3 + 3 design and the continuous reassessment method. Results: By accounting for the risk-benefit trade-off, the EffTox phase I-II design overcomes the limitations of conventional toxicity-based phase I designs. Numerical simulations show that the EffTox design has higher probabilities of identifying the optimal dose and treats more patients at the optimal dose. Conclusions: Phase I-II designs, such as the EffTox design, provide a coherent and efficient approach to finding the optimal P2RD by explicitly accounting for risk-benefit trade-offs underlying medical decisions.

Comparison of Mitochondrial Function in Boar and Bull Spermatozoa Throughout Cryopreservation Based on JC-1 Staining.

BACKGROUND: Poor reproductivity hampers the commercialization of cryopreserved boar semen. OBJECTIVE: This study was to determine the differences in the sperm mitochondrial function between boar and bull semen at different cryopreservation stages. MATERIALS AND METHODS: Boar and bull fresh, equilibrated, and frozen-thawed spermatozoa were evaluated for mitochondrial function using JC-1 under a fluorescent microscope. RESULTS: Bull and boar percentage of spermatozoa staining green (PSSG) showed no difference between fresh and equilibrated semen (P> 0.05). However, frozen-thawed bull and boar semen demonstrated significantly higher PSSG (P < 0.01) than fresh and equilibrated semen. Frozen-thawed boar semen represented a significantly higher PSSG (P < 0.01) than bull semen. CONCLUSION: Negative cryopreservation influence on boar and bull spermatozoa was not significantly produced by pre-freezing procedures, but rather by freezing and thawing. Cryopreservation has more pronounced negative effects on boar than on bull spermatozoa, which partly explains lagged commercialization of frozen boar semen.

Derivation and characterization of primordial germ cells from Guangxi yellow-feather chickens.

The Guangxi yellow-feather chicken is a very important breed used as a broiler in southern China, but the pure line is being threatened by continual introduction of foreign genetics into its breeding program to make it more marketable. In the current study, we isolated primordial germ cells (PGCs), a cell type committed to form sperm or eggs and that is responsible for passing genetic material from one generation to the next, from Guangxi yellow-feather chickens and cultured them in a cell-insert system. Three stable cell lines, all male, were established from 10 isolations. These cells proliferated and expressed germ cell-related markers such as SSEA-1 and EMA-1 after prolonged culture in vitro. After genetic modification, these PGCs retained significant potential to colonize the gonads and give rise to gametes when they were reintroduced into the vasculature of stage-15 HH embryos, confirming their germline cell characteristics. The ability to culture PGCs and preserve the genetics from this species would not only be of significant importance for biodiversity conservation, but also holds promise for use of these cells in breeding strategies in the future.

Isolation, proliferation, and induction of Bama mini-pig spermatogonial stem cells in vitro.

Spermatogonial stem cells (SSCs), the unique seed cells of testes, can undergo meiosis and form spermatozoa, thus transmitting genetic information to offspring. Research concerning these cells explores the mechanism underlying spermatogenesis, making possible the induction of their differentiation into spermatozoa in vitro. SSCs have therefore attracted much interest among scientists. Although the proliferation of such cells in vitro has been demonstrated, we are unaware of any long-term laboratory culture of porcine SSCs. The objective of this study was to isolate, characterize, culture, and induce the differentiation of Bama mini-pig SSCs. SSCs were isolated using differential plating and cultured for over 100 days on an STO feeder cell layer without serum. Cell clusters appeared after three passages and continuously formed during subsequent cultivation. Staining showed that these clusters were positive for UCHL1 and CDH1, could be bound by Dolichos biflorus agglutinin, and that some cells expressed OCT4. Ultrastructure observations revealed SSCs in testis tissue to be round in shape, while those cultured in vitro were flat and bound together. Our attempts at inducing differentiation showed that SSCs cultured in vitro could undergo meiosis. In this study, we describe an effective culture system for Bama mini-pig SSCs capable of producing enough cells to establish a platform for further SSC research, such as genetic manipulation or exploration of the mechanism underlying spermatogenesis.

Stability of the cytoskeleton of matured buffalo oocytes pretreated with cytochalasin B prior to vitrification.

Stabilizing the cytoskeleton system during vitrification can improve the post-thaw survival and development of vitrified oocytes. The cytoskeleton stabilizer cytochalasin B (CB) has been used in cryopreservation to improve the developmental competence of vitrified oocytes. To assess the effect of pretreating matured buffalo oocytes with CB before vitrification, we applied 0, 4, 8, or 12 µg/mL CB for 30 min. The optimum concentration of CB treatment (8 µg/mL for 30 min) was then used to evaluate the distribution of microtubules and microfilaments, the expression of the cytoskeleton proteins actin and tubulin, and the developmental potential of matured oocytes that were vitrified-warmed by the Cryotop method. Western blotting demonstrated that vitrification significantly decreased tubulin expression, but that the decrease was attenuated for oocytes pretreated with 8 µg/mL CB before vitrification. After warming and intracytoplasmic sperm injection, oocytes that were pretreated with 8 µg/mL CB before vitrification yielded significantly higher 8-cell and blastocyst rates than those that were vitrified without CB pretreatment. The values for the vitrified groups in all experiments were significantly lower (P < 0.01) than those of the control groups. In conclusion, pretreatment with 8 µg/mL CB for 30 min significantly improves the cytoskeletal structure, expression of tubulin, and development capacity of vitrified matured buffalo oocytes.

Loss of p27 Associated with Risk for Endometrial Carcinoma Arising in the Setting of Obesity.

Endometrial carcinoma (EC) exhibits the strongest association with obesity of all cancers. Growth of these tumors is driven by PI3K/AKT activation, and opposed by tumor suppressors, including the tuberous sclerosis complex 2 (TSC-2) and p27, with inactivation of TSC2 and loss or cytoplasmic mislocalization of p27 both being linked to PI3K/AKT activation. However, little is known about the involvement of p27 in the development of EC arising in the setting of obesity, especially its role early in disease progression. Using a panel of EC cell lines, in vitro studies using PI3K inhibitors provided evidence that p27 rescue contributes to the efficacy of interventions that inhibit endometrial cell growth. In "at risk" obese patients, and in an animal model of obesity-associated EC (Tsc2-deficient Eker rats), p27 was moderately-to-severely reduced in both "normal" endometrial glands as well as in endometrial complex atypical hyperplasia (obese women), and endometrial hyperplasia (obese rats). In obese Eker rats, an energy balance intervention; caloric restriction from 2-4 months of age, reduced weight, increased adiponectin and lowered leptin to produce a favorable leptin:adiponectin ratio, and reduced circulating insulin levels. Caloric restriction also increased p27 levels, relocalized this tumor suppressor to the nucleus, and significantly decreased hyperplasia incidence. Thus, dietary and pharmacologic interventions that inhibit growth and decrease risk for development of endometrial lesions are associated with increased expression and nuclear (re)localization of p27. These data suggest that p27 levels and localization may be useful as a biomarker, and possible determinant, of risk for EC arising in the setting of obesity.

Production of hGFAP-DsRed transgenic Guangxi Bama mini-pigs via somatic cell nuclear transfer.

The mini-pig is a useful animal model for human biomedical research due to its physiological similarity to humans and the ease of handling. In order to optimize the efficiency of production of transgenic Bama mini-pigs through somatic cell nuclear transfer (SCNT), we examined the effects of contact inhibition, roscovitine treatment, and serum starvation on the cell cycle synchronization and transgenic cloned embryo development in vivo and in vitro after nuclear transfer. The analysis showed that the rates of G0/G1 stage cells in the contact inhibition (92.11%) and roscovitine treatment groups (89.59%) were significantly higher than in serum starvation group (80.82%). A higher rate of apoptosis was seen in the serum starvation group (14.13%) compared to the contact inhibition and roscovitine treatment groups (6.71 and 2.46% respectively, P < 0.05). There was a significant decrease in blastocyst yield in the serum starvation group (14.19%) compared to the roscovitine treatment and contact inhibition groups (21.31 and 20.32% respectively, P < 0.05). A total of 1070 transgenic cloned embryos derived from the three treatment groups were transferred to surrogate sows; one pregnancy was established and three embryos from the roscovitine treatment group successfully completed gestation. These results indicate that the roscovitine treatment was more effective at synchronizing transgenic kidney cells in Bama mini-pigs and allowed reconstructed embryos to develop to full term.

EZH2-mediated epigenetic suppression of long noncoding RNA SPRY4-IT1 promotes NSCLC cell proliferation and metastasis by affecting the epithelial-mesenchymal transition.

Recent evidence indicates that long noncoding RNAs (lncRNAs) have a critical role in the regulation of cellular processes such as differentiation, proliferation, and metastasis. These lncRNAs are dysregulated in a variety of cancers and many function as tumor suppressors; however, the regulatory factors involved in silencing lncRNA transcription are poorly understood. In this study, we showed that epigenetic silencing of lncRNA SPRY4 intronic transcript 1 (SPRY4-IT1) occurs in non-small-cell lung cancer (NSCLC) cells through direct transcriptional repression mediated by the Polycomb group protein enhancer of zeste homolog 2 (EZH2). SPRY4-IT1 is derived from an intron within SPRY4, and is upregulated in melanoma cells; knockdown of its expression leads to cell growth arrest, invasion inhibition, and elevated rates of apoptosis. Upon depletion of EZH2 by RNA interference, SPRY4-IT1 expression was restored, and transfection of SPRY4-IT1 into NSCLC cells resulted in a significant antitumoral effect, both in culture and in xenografted nude mice. Moreover, overexpression of SPRY4-IT1 was found to have a key role in the epithelial-mesenchymal transition through the regulation of E-cadherin and vimentin expression. In EZH2-knockdown cells, which characteristically showed impaired cell proliferation and metastasis, the induction of SPRY4-IT1 depletion partially rescued the oncogenic phenotype, suggesting that SPRY4-IT1 repression has an important role in EZH2 oncogenesis. Of most relevance, translation of these findings into human NSCLC tissue samples demonstrated that patients with low levels of SPRY4-IT1 expression had a shorter overall survival time, suggesting that SPRY4-IT1 could be a biomarker for poor prognosis of NSCLC.

Response to an exercise intervention after endometrial cancer: differences between obese and non-obese survivors.

OBJECTIVE: The objective of this paper is to describe baseline differences between obese and non-obese endometrial cancer survivor in anthropometrics, exercise behavior, fitness, heart rate and blood pressure, and quality of life, and to analyze whether the effect of a home-based exercise intervention on these outcomes differed for obese and non-obese participants. METHODS: One hundred post-treatment Stage I-IIIa endometrial cancer survivors participated in a single arm 6month study in which they received a home-based exercise intervention. Cardiorespiratory fitness, anthropometrics, and exercise behavior were measured every two months, and quality of life (QOL) and psychological distress were measured at baseline and 6months. RESULTS: Adjusting for potential confounders, at baseline obese survivors had poorer cardiorespiratory fitness (p=.002), higher systolic blood pressure (p=.018), and lower physical functioning (p<.001) and ratings of general health (p=.002), and more pain (p=.037) and somatization (.002). Significant improvements were seen in exercise behavior, resting heart rate, systolic blood pressure, and multiple QOL domains over the course of the intervention. Obese survivors had less improvement in exercise behavior and cardiorespiratory fitness than non-obese survivors, but there were no differences with regard to improvements in QOL and stress. CONCLUSIONS: Home based exercise interventions are beneficial to endometrial cancer survivors, including those whose BMI is in the obese range. While obese survivors have lower levels of physical activity and fitness, they experienced similar activity, fitness, quality of life and mental health benefits. Exercise should be encouraged in endometrial cancer survivors, including those who are obese.

Gynecologic cancer screening and communication with health care providers in women with Lynch syndrome.

We evaluated knowledge of gynecologic cancer screening recommendations, screening behaviors, and communication with providers among women with Lynch syndrome (LS). Women aged ≥25 years who were at risk for LS-associated cancers completed a semi-structured interview and a questionnaire. Of 74 participants (mean age 40 years), 61% knew the appropriate age to begin screening, 75-80% correctly identified the recommended screening frequency, and 84% reported no previous screening endometrial biopsy. Women initiated discussions with their providers about their LS cancer risks, but many used nonspecific terms or relied on family history. Most were not offered high-risk screening options. While many women were aware of risk-appropriate LS screening guidelines, adherence was suboptimal. Improving communication between women and their providers regarding LS-related gynecologic cancer risk and screening options may help improve adherence.

The Oklahoma Thrombotic Thrombocytopenic Purpura-haemolytic Uraemic Syndrome Registry. A model for clinical research, education and patient care.

The Oklahoma Thrombotic Thrombocytopenic Purpura-Haemolytic Uraemic Syndrome (TTP-HUS) Registry has a 24 year record of success for collaborative clinical research, education, and patient care. This article tells the story of how the Registry began and it describes the Registry's structure and function. The Registry provides a model for using a cohort of consecutive patients to investigate a rare disorder. Collaboration between Oklahoma, United States and Bern, Switzerland has been the basis for successful interpretation of Registry data. Registry data have provided new insights into the evaluation and management of TTP. Because recovery from acute episodes of TTP has been assumed to be complete, the increased prevalence of hypertension, diabetes, depression, and death documented by long-term follow-up was unexpected. Registry data have provided opportunities for projects for students and trainees, education of physicians and nurses, and also for patients themselves. During our follow-up, patients have also educated Registry investigators about problems that persist after recovery from an acute episode of TTP. Most important, Registry data have resulted in important improvements for patient care.

Patterns of genomic loss of heterozygosity predict homologous recombination repair defects in epithelial ovarian cancer.

BACKGROUND: Defects in BRCA1, BRCA2, and other members of the homologous recombination pathway have potential therapeutic relevance when used to support agents that introduce or exploit double-stranded DNA breaks. This study examines the association between homologous recombination defects and genomic patterns of loss of heterozygosity (LOH). METHODS: Ovarian tumours from two independent data sets were characterised for defects in BRCA1, BRCA2, and RAD51C, and LOH profiles were generated. Publically available data were downloaded for a third independent data set. The same analyses were performed on 57 cancer cell lines. RESULTS: Loss of heterozygosity regions of intermediate size were observed more frequently in tumours with defective BRCA1 or BRCA2 (P=10(-11)). The homologous recombination deficiency (HRD) score was defined as the number of these regions observed in a tumour sample. The association between HRD score and BRCA deficiency was validated in two independent ovarian cancer data sets (P=10(-5) and 10(-29)), and identified breast and pancreatic cell lines with BRCA defects. CONCLUSION: The HRD score appears capable of detecting homologous recombination defects regardless of aetiology or mechanism. This score could facilitate the use of PARP inhibitors and platinum in breast, ovarian, and other cancers.

Platelet-rich fibrin increases cell attachment, proliferation and collagen-related protein expression of human osteoblasts.

BACKGROUND: Platelet-rich fibrin (PRF) prepared by Choukroun's technique is derived from an autogenous preparation of concentrated platelets without any manipulation. PRF was found to increase osteoblast growth and proliferation. However, the underlying mechanisms are not yet completely understood. This study aimed to determine the effects of PRF on cell attachment, proliferation, phosphorylated Akt, heat shock protein 47 (HSP47) and lysyl oxidase (LOX) expression on human osteoblasts. METHODS: Blood collection was carried out from 10 healthy volunteers. Cell attachment and proliferation were measured by colorimetric assay with WST-1 and alamar blue in human osteoblast cell line U2OS cells, respectively. Western blot was employed to evaluate the expression of p-Akt, HSP47 and LOX. RESULTS: PRF alone was found to stimulate U2OS cell attachment compared with untreated controls (p < 0.05). PRF was found to increase osteoblast proliferation during a 5-day incubation period (p < 0.05). PRF was found to increase Akt phosphorylation in a time-dependent manner (p < 0.05). Collagen-related proteins HSP47 and LOX were significantly elevated by stimulation with PRF compared with untreated controls (p < 0.05). CONCLUSIONS: It is suggested that PRF is capable of increasing osteoblast attachment, proliferation and simultaneously upregulating collagen-related protein production. These actions in combination would effectively promote bone regeneration.

Translocation of active mitochondria during buffalo (Bubalus bubalis) oocytes in vitro maturation, fertilization and preimplantation embryo development.

Mitochondria are energy-supplying organelles, whose distribution and functional integrity are necessary for cell survival and development. In this study, the mitochondrial distribution pattern and activity during buffalo oocyte in vitro maturation, fertilization and preimplantation embryo development were revealed using a fluorescent dye and confocal laser scanning microscopy. Distribution of active mitochondria changed during buffalo oocyte in vitro maturation. Active mitochondria were transferred from the outer to inner and perinuclear cytoplasm as oocytes matured in vitro and aggregated around the pronuclei in the fertilized eggs. Active mitochondria were also observed in preimplantation embryos. In the two-cell stage, they were distributed throughout the cytoplasm. From four-cell to the spherical embryonic stages, active mitochondria translocated to the perinuclear and the periphery of the cytoplasm. These results confirm that mitochondria play an important role in oocyte and embryo. The distribution of active mitochondria might be a marked feature of buffalo oocyte maturation, fertilization and preimplantation embryo development in vitro.

Protective effects of melatonin against oxidative stress in flow cytometry-sorted buffalo sperm.

Previous reports of the ability of melatonin to scavenge a variety of toxic oxygen and nitrogen-based reactants suggest that melatonin could be an effective antioxidant for protecting sperm. In this study, flow cytometry and laser tweezers Raman spectroscopy were used to evaluate the effect of melatonin on buffalo sperm quality to optimize sperm sex-sorting procedures. In fresh sperm incubated in the presence or absence of melatonin (10(-4) m) for 1, 24, 48 h or 72 h at 27°C, the mitochondrial activity was significantly higher than in a non-melatonin control (p < 0.05). Also, during the flow-sorting process, sperm in melatonin-supplemented groups had higher (p < 0.05) mitochondrial activity than the control. The intensity of Raman spectra from sperm frozen in media supplemented with melatonin was significantly weaker than that for non-melatonin-treated groups, except for a band at 1302 per cm. Thus, melatonin helps to protect buffalo sperm from reactive oxygen species induced by staining, sorting and freezing and increases semen quality after the freezing-thawing processes. Furthermore, the results indicate the high potential of the laser tweezers Raman spectroscopy technique for rapid, effective and non-invasive assessment of the quality of sperm cells.

Communication of BRCA1 and BRCA2 genetic test results to health care providers following genetic testing at a tertiary care center.

Individuals at high risk for hereditary cancers often receive genetic counseling and testing at tertiary care centers; however, they may receive care for long-term management of their cancer risk in community settings. Communication of genetic test results to health care providers outside of tertiary care settings can facilitate the long-term management of high risk individuals. This study assessed women's communication of BRCA1/BRCA2 genetic test results to health care providers outside of tertiary care settings (termed "outside" health care providers, or OHCPs) and women's perceptions regarding communication of results. Women (n = 312) who underwent BRCA1/BRCA2 genetic counseling and testing completed a questionnaire assessing whether or not they shared test results with OHCPs and perceptions regarding the communication of test results to OHCPs. Most (72%) shared genetic test results with OHCPs. Women with no personal history of cancer were more likely to have shared results compared to women with a personal history of cancer. Mutation status did not significantly predict sharing of genetic information. Most reported positive perceptions regarding the disclosure of genetic test results to OHCPs. The majority did not report any concerns about potential insurance discrimination (88%) and indicated that OHCPs were able to appropriately address their questions (81%). Although most women shared their genetic test results with OHCPs, those with a personal history of cancer may need further encouragement to share this information. Tertiary care centers should facilitate outreach and education with OHCPs in order to assure appropriate long-term cancer risk management for high risk populations.