Outcomes of Dual Mobility Bearings in Revision Total Hip Replacements.

Background Dual mobility bearings have gained attention in the prevention of instability in revision total hip replacement. This study aimed to evaluate the use of dual mobility bearings in revision total hip replacement. The primary outcome was the rate of dislocation. Secondary outcomes included the rate of re-operation for any reason, surgical complications, serious medical adverse events, and 90-day mortality rate. Methods A single-centre case series of 55 consecutive operations in 49 patients who underwent revision total hip replacement using dual mobility bearings with a minimum follow-up of three months was studied.  Results Early dislocation occurred in one case (2%), and there were no intra-prosthetic dislocations at a mean follow-up of 16 months. The rate of re-operation for any reason was 6/55 (11%) cases, and the post-operative infection rate was 2/55 (4%) cases. Serious medical adverse events occurred in 2/55 (4%) cases. The 90-day mortality rate was 1/55 (2%) cases. Two cases (2%) had cup abduction or anteversion angles outside of the safe zones although there were no dislocations in these patients. Conclusion This case series demonstrates a low dislocation rate in the early post-operative period for dual mobility bearings in revision total hip replacement. Dual mobility bearings show promise as an early low dislocation implant in revision total hip replacement. It remains to be determined whether dual mobility bearings are low-wear implants in the long term.

Investigation of the effects of thymoquinone (TQ) and polyvinylpyrrolidone (PVP) on the motility, viability, and chromatin status of sperm in normozoospermic semen samples.

Objective: Adverse effects of polyvinylpyrrolidone (PVP) on sperm membrane and chromatin have been proven in many studies. Among the natural products proposed as an alternative for PVP, thymoquinone (TQ)-a major constituent of Nigella sativa plant- has been suggested as a potential natural spermostasis. Therefore, this study aimed to compare the effects of TQ with PVP for sperms motility, survival, DNA denaturation, and DNA fragmentation in normozoospermic men (men with a normal or healthy sperm profile). Methods: An experimental trial was carried out on 30 normozoospermic men of the Andrology Unit of (Shahid Beheshti Hospital, Isfahan, Iran). Each washed semen samples were divided into four fractions and was randomly treated with TQ (50 µg/ml), %5 PVP, and %10 PVP (M/V) which was compared to untreated fraction (control). Results: There was a significant difference between the four groups in terms of motility, viability, DNA denaturation, and fragmentation (P <0.05). TQ caused sperm immobility, while 5% PVP and 10% PVP decreased (98 and 99%, respectively) sperm motility compared to control. TQ did not affect sperm viability compared to the control group, but PVP decreased it. Besides, TQ did not affect DNA denaturation and fragmentation, but PVP increased it. Conclusion: TQ could be used as an alternative natural spermostasis with less adverse effects rather than PVP which causes more efficient immobilization and isolation of individual sperm cells.

Subsidence in Centraliser Sign: A Novel Tool to Help Predict Early Subsidence in Periprosthetic Femoral Fractures Around Polished Tapered Stems Treated With Internal Fixation.

Background When treating periprosthetic femoral fractures (PPF) around polished taper slip stems (PTS), determining which patients can be successfully treated with internal fixation can be challenging. We have described the subsidence-in-centraliser (SINC) sign as a radiographic feature of PPF around PTS stems. We hypothesise that a positive SINC sign can help predict a poorer outcome for the fixation of these fractures. Patients and methods Retrospective identification of PPFs around cemented PTS with an appreciable centraliser on radiographs was conducted at a single centre. A positive SINC sign was defined as a post-injury radiograph demonstrating >50% reduction in the radiographic lucency representing the stem centraliser when compared to pre-injury films or complete obliteration of distal lucency when no pre-injury film was available. The primary outcome was the rate of subsequent stem subsidence on follow-up radiographs comparing SINC-positive and SINC-negative fractures, which were managed with open reduction and internal fixation (ORIF). Results Fifty-four patients were included in the analysis. The mean age was 76.8 years, and the mean follow-up for all patients was 12.7 months. Thirty-five fractures were deemed SINC-positive, and 19 were SINC-negative. 17/17 (100%) SINC-positive fractures managed with fixation underwent further subsidence (mean 5.4 mm, SD 2.8). A positive SINC sign demonstrated a sensitivity of 90.5% and specificity of 100% for subsequent stem subsidence in fractures treated without revision. SINC positive fractures underwent significantly more subsidence compared with SINC negative fractures when fixed (5.4 mm vs. 0.28 mm, U = 6.50, p<0.001) at a mean follow-up of 12.7 months. The SINC sign demonstrated strong inter- (k=0.96) and intra-rater (k=0.86) reliability. Conclusion The SINC sign can serve as a useful adjunct in the decision to fix or revise PPF around PTS. A positive SINC sign may represent a cement mantle that cannot be reconstituted anatomically, leading to subsidence after treatment with ORIF.

Accuracy of Total Hip Arthroplasty Templating Using Set Calibration Magnifications.

Background Templating for total hip arthroplasty has been adopted over recent decades as a reliable and accurate method for pre-operative planning. The use of calibration markers for this process provides a recognised benefit at the expense of cost, availability and error. Many surgeons use a set magnification of 118% to account for calibration errors when templating total hip arthroplasty. This study aims to assess the accuracy of templating with standardised magnifications and assess the effect of BMI on templating accuracy. Materials and methods A retrospective analysis was performed using a single-surgeon series of 119 consecutive total hip arthroplasties. Anteroposterior radiographs were taken pre- or post-operatively without calibration hardware. Pre-operatively, the total hip arthroplasty was templated on TraumaCad (BrainLab Inc, Westchester, IL) using either 118% or 119% calibration magnification. Post-operative magnification was calibrated using the known femoral head diameter. Templated and implanted prostheses were compared for size. Results At 118%, 61.1% of cups matched those templated with 96.3% of cups within two sizes. At 119%, 52.5% of cups used matched their templates with 100% within two sizes. There was no significant difference between 118% and 119% cup size prediction (p=0.49). A trend was noticed in increasing magnification error with increasing BMI. However, BMI had no significant effect on the accuracy of templating cup size within two cup sizes (p=0.58). Conclusion. Templating acetabular cups using a set magnification of 118% or 119% yields accurate results and provides a reliable method to template without calibration equipment. Whilst BMI can affect magnification error, this has no significant effect on the accuracy of implanted cups and stems within two sizes.

Telemedicine in orthopaedics during COVID-19 and beyond: A systematic review.

INTRODUCTION: Telemedicine is the delivery of healthcare across a distance using some form of communication technology. The COVID-19 pandemic has led to increased adoption of telemedicine with national orthopaedic governing bodies advocating its use, as evidence suggests that social distancing maybe necessary until 2022. This systematic review aims to explore evidence for telemedicine in orthopaedics to determine its advantages, validity, effectiveness and utilisation. METHODS: Databases of PubMed, Web of Science, Scopus and CINAHL were systematically searched and articles were included if they involved any form of telephone or video consultation in an orthopaedic population. Findings were synthesised into four themes: patient/clinician satisfaction, accuracy and validity of examination, safety and patient outcomes and cost effectiveness. Quality assessment was undertaken using Cochrane and Joanna Briggs Institute appraisal tools. RESULTS: Twenty-one studies were included consisting of nine randomised controlled trials (RCTs). Studies revealed high patient satisfaction with telemedicine for convenience, less waiting and travelling time. Telemedicine was cost effective particularly if patients had to travel long distances, required hospital transport or time off work. No clinically significant differences were found in patient examination nor measurement of patient-reported outcome measures. Telemedicine was reported to be a safe method of consultation. DISCUSSION: Evidence suggests that telemedicine in orthopaedics can be safe, cost effective, valid in clinical assessment and with high patient/clinician satisfaction. However, more high-quality RCTs are required to elucidate long-term outcomes. This systematic review presents up-to-date evidence on the use of telemedicine and provides data for organisations considering its use during the COVID-19 pandemic and beyond.

Fixation and Implant Retention of Extracapsular Femoral Neck Periprosthetic Fractures Around Hip Resurfacing Arthroplasty - A Case Series.

BACKGROUND: Extracapsular femoral neck fractures in the presence of a resurfacing hip arthroplasty (RHA) appear to be independent of suboptimal technique during the initial implantation of the RHA and present with a similar etiology as native hip fractures - that is, a fragility fracture related to pathological or age-related osteoporosis, as a consequence of trauma. In the presence of a well-fixed and previously well-functioning RHA, the options for management include revision arthroplasty or open reduction and internal fixation (ORIF). In the absence of loosening through mechanisms of wear, infection, metallosis, or suboptimal prosthesis positioning, many authors have advocated ORIF with implant retention. However, there is often debate regarding the use of total hip arthroplasty in these cases. CASE SERIES: The authors conducted a thorough assessment of the literature followed by a retrospective review of outcomes for three patients treated by ORIF with implant retention for extracapsular femoral neck fractures around a RHA, using a standardized technique. All patients were independently mobile and active with well-fixed and well-functioning RHAs before the date of injury. All patients suffered low-energy trauma resulting in the fracture. There were no intraoperative or perioperative complications. All patients achieved full weight-bearing status and independent mobility. Two patients achieved radiographic union and returned to full range of movement and independent mobilization comparable to their preoperative state. One patient was lost to follow-up. CONCLUSION: The authors believe that fixation of extracapsular proximal femoral fractures distal to a well-fixed, well-functioning RHA is a good management option in an independent and active patient. A higher level of evidence is needed to investigate the surgical management options of these injuries comparing osteosynthesis with revision arthroplasty.

Bilateral osteochondral defects of the patellae in an 11-year-old girl.

Osteochondral defect or osteochondritis dissecans (OCD) of the knee usually affects young, active populations. It is a challenging diagnosis as patients typically present with poorly localised activity-related pain, which is non-specific and covers many differentials. We present an active 11-year-old girl with bilateral osteochondral defects of the patellae: a rare clinical disorder which was affecting her sporting activities. She had a 12-month history of bilateral anterior knee pain before the diagnosis was achieved with appropriate imaging. Her pain significantly improved with activity modification and physiotherapy. Follow-up will require outpatient clinic assessment and imaging to determine if non-operative management continues to be successful or surgery may be required. This case report emphasises the importance of appropriate high index of suspicion when managing patients with non-specific knee pain. It also demonstrates the importance of judicious use of imaging to avoid a missed or delayed diagnosis.

Effects of temperature and storage time on the motility, viability, DNA integrity and apoptosis of processed human spermatozoa.

The aim of this study was to evaluate motility, viability, DNA integrity and apoptosis of spermatozoa when washed semen samples were kept for up to 12 days at 4-6°C and 25°C. In this experimental study, 26 normozoospermic semen samples were washed twice in Modified Ham's F10 and resuspended in IVF fertilisation medium. Half of the specimens were stored at 4-6°C, and the other half was kept at 25°C for 12 days. The proportions of viable, motile, spermatozoa with double-stranded DNA and apoptotic spermatozoa were examined during storage time. Apoptosis was measured using annexin V-PI staining followed by flow cytometry. Results showed that sperm motility and viability decreased during 12 days of sample storage (p < .001). There was no significant difference between the two temperatures in terms of motility and viability for up to 2 days (p < .05). The percentage of spermatozoa with double-stranded DNA remained unchanged during the 12 days of storage at both temperatures (p > .05). Although there was no difference between the two temperatures in terms of motility, viability and apoptosis during the first two days of storage, storage of spermatozoa at 4-6°C is better than storage for a longer period than storage at 25°C. Sperm DNA resisted against denaturation during storage.

Cement Pedestal Spacer Technique for Infected Two-stage Revision Knee Arthroplasty: Description and Comparison of Complications.

BACKGROUND: Infection following total knee arthroplasty (TKA) is a significant complication, with an incidence of up to 2% in primary TKA and 4%-8% in revision cases. Two-stage revision is the gold standard treatment for long-lasting infections of TKA. The purpose of this study was to describe the cement pedestal spacer technique used in infected two-stage revision knee arthroplasty and compare complications against conventional fixed and mobile cement spacers. PATIENTS AND METHODS: A retrospective review was conducted in all cases who underwent two-stage TKA revision for infection between 2009 and 2015. These cases were separated into groups depending on the cement spacer utilized (fixed, mobile nonpedestal, and mobile spacers with cement pedestal). The cement pedestal technique involves press fitting a cement cylinder into the femur before definitive spacer insertion. RESULTS: Forty four patients underwent two-stage revision TKA. Fewest complications were observed in the pedestal group, with no spacers having subluxed/tilted. The longest followup was also observed in the pedestal group (mean 52.5 months). Mobile spacers with no cement pedestal displayed the highest reinfection rate (16.7%) and the greatest number of cases with complications (malalignment, subluxation, tilting, and spacer fracture). All patients in the pedestal group were ambulatory after the first-stage revision. CONCLUSIONS: The cement pedestal technique minimizes complications by optimizing component positioning and balancing. It also safely extends the indication for an articulated spacer into a set of cases with more extensive bone loss and allows for extended monitoring of inflammatory markers.

Correlation between sperm motility and sperm chromatin/DNA damage before and after cryopreservation and the effect of folic acid and nicotinic acid on post-thaw sperm quality in normozoospermic men.

Cryopreservation exposes sperm to physical and chemical stresses causing cell damages and impairs sperm functions. The aim of this study was to evaluate the association between motility and sperm chromatin/DNA damage before and after cryopreservation and investigate the effects of folic acid and nicotinic acid on post-thaw sperm quality. Thirty semen samples were obtained from 30 normozoospermic men, aged between 25 and 45 years old. Each sample were divided into five aliquots to form the following groups: fresh, cryopreserved with sperm-freeze only (control), with nicotinic acid (10 mM), with folic acid (50 nM), and with a combination of folic acid (50 nM) + nicotinic acid (10 mM). Sperm viability and motility in each group were assessed by eosin-nigrosine staining and computer-aided sperm analysis respectively. Sperm chromatin quality was studied by aniline blue, toluidine blue, acridine orange staining methods and sperm chromatin dispersion test. Cryopreservation led to a significant reduction in sperm quality in comparison to fresh sample groups (p < 0.05). Sperm chromatin damage was negatively correlated with the percentage of progressively motile cells. Supplementation of the cryopreservation medium with folic acid or nicotinic acid induced a significant improvement in sperm parameters and chromatin quality, compared to control groups (p < 0.05). Meanwhile, the combination of folic acid + nicotinic acid showed a significant protective effect in post thaw sperm. In conclusion, cryopreservation generated oxidative stress, inducingsperm cryodamage, reducing progressive motility and sperm quality, as an indicator of significant chromatin/DNA damage. Folic acid and nicotinic acid exhibited a potential cryoprotective effect by enhancing sperm quality.

Can an Augmented Reality Headset Improve Accuracy of Acetabular Cup Orientation in Simulated THA? A Randomized Trial.

BACKGROUND: Accurate implant orientation reduces wear and increases stability in arthroplasty but is a technically demanding skill. Augmented reality (AR) headsets overlay digital information on top of the real world. We have developed an enhanced AR headset capable of tracking bony anatomy in relation to an implant, but it has not yet been assessed for its suitability as a training tool for implant orientation. QUESTIONS/PURPOSES: (1) In the setting of simulated THA performed by novices, does an AR headset improve the accuracy of acetabular component positioning compared with hands-on training by an expert surgeon? (2) What are trainees' perceptions of the AR headset in terms of realism of the task, acceptability of the technology, and its potential role for surgical training? METHODS: Twenty-four study participants (medical students in their final year of school, who were applying to surgery residency programs, and who had no prior arthroplasty experience) participated in a randomized simulation trial using an AR headset and a simulated THA. Participants were randomized to two groups completing four once-weekly sessions of baseline assessment, training, and reassessment. One group trained using AR (with live holographic orientation feedback) and the other received one-on-one training from a hip arthroplasty surgeon. Demographics and baseline performance in orienting an acetabular implant to six patient-specific values on the phantom pelvis were collected before training and were comparable. The orientation error in degrees between the planned and achieved orientations was measured and was not different between groups with the numbers available (surgeon group mean error ± SD 16° ± 7° versus AR 14° ± 7°; p = 0.22). Participants trained by AR also completed a validated posttraining questionnaire evaluating their experiences. RESULTS: During the four training sessions, participants using AR-guidance had smaller mean (± SD) errors in orientation than those receiving guidance from the surgeon: 1° ± 1° versus AR 6° ± 4°, p < 0.001. In the fourth session's assessment, participants in both groups had improved (surgeon group mean improvement 6°, 95% CI, 4-8°; p < 0.001 versus AR group 9°, 95% CI 7-10°; p < 0.001). There was no difference between participants in the surgeon-trained and AR-trained group: mean difference 1.2°, 95% CI, -1.8 to 4.2°; p = 0.281. In posttraining evaluation, 11 of 12 participants would use the AR platform as a training tool for developing visuospatial skills and 10 of 12 for procedure-specific rehearsals. Most participants (11 of 12) stated that a combination of an expert trainer for learning and AR for unsupervised training would be preferred. CONCLUSIONS: A novel head-mounted AR platform tracked an implant in relation to bony anatomy to a clinically relevant level of accuracy during simulated THA. Learners were equally accurate, whether trained by AR or a surgeon. The platform enabled the use of real instruments and gave live feedback; AR was thus considered a feasible and valuable training tool as an adjunct to expert guidance in the operating room. Although there were no differences in accuracy between the groups trained using AR and those trained by an expert surgeon, we believe the tool may be useful in education because it demonstrates that some motor skills for arthroplasty may be learned in an unsupervised setting. Future studies will evaluate AR-training for arthroplasty skills other than cup orientation and its transfer validity to real surgery. LEVEL OF EVIDENCE: Level I, therapeutic study.

Effect of preservation of human semen sample at 4-6 and 25 °C on sperm motility.

Sperm motility is the result of transverse movements that exist along its tail. It plays an important role in male fertility. The aim of this study was to evaluate the influence of keeping washed normozoospermic semen samples at 4-6 and 25 °C on the motility of spermatozoa. 26 semen samples of normozoospermic were washed twice in modified Ham's F10 medium. Then, thirteen of the semen samples were kept in refrigerator (4-6 °C) and the remaining samples were stored in incubator (25 °C) for 12 days. On the 0 (immediately after sampling as control group), 1st, 2nd, 5th, 7th and the 12th days, the percentage of fast progressive (grade a), slow progressive (grade b), non-progressive (grade c) and immotile (grade d) sperm cells were calculated for each temperature. The data obtained from this study showed that the percentages of a, b and c grades of motile spermatozoa were significantly decreased (p < 0.001) during 12 days at the both temperatures but reduction of these percentages has a gentle slope at 4-6 °C. There was no motile sperm after 12 days of storage. This study suggests that motile spermatozoa could be retrieved up to 7 days after the storage of washed normozoospermic men semen samples at 4-6 and 25 °C. Also, there were no motile sperm cells 12 days after sampling.

Comparison of the efficacy of piascledine and transforming growth factor ß1 on chondrogenic differentiation of human adipose-derived stem cells in fibrin and fibrin-alginate scaffolds.

OBJECTIVES: The aim of this study was to compare the chondrogenic induction potential of Piascledine and TGF-ß1 on adipose-derived stem cells (ADSCs) in fibrin and fibrin-alginate scaffolds. MATERIALS AND METHODS: Human subcutaneous adipose tissues were harvested from three patients who were scheduled to undergo liposuction. Isolated ADSCs were proliferated in a culture medium. Then, the cells were seeded in fibrin or fibrin-alginate scaffolds and cultured for 14 days in a chondrogenic medium containing Piascledine, TGF-ß1, or both. The rate of cell proliferation and survival was evaluated by using MTT [3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide] assay and the rate of the expression of type II collagen, aggrecan, and type X collagen genes was evaluated by real-time polymerase chain reaction (real-time PCR) method. RESULTS: The MTT results showed that Piascledine is able to enhance the proliferation and survival of ADSCs in fibrin scaffolds in comparison to other groups (P<0.05). Real-time PCR evaluation revealed that the expression of type II collagen was higher in TGF- ß1groups, but the expression of aggrecan was higher in TGF-ß1 alone or along with Piascledine in fibrin-alginate scaffolds. Furthermore, the expression of type X collagen was lower in Piascledine alone or along with TGF-ß1 in fibrin scaffold. CONCLUSION: Piascledine can enhance the proliferation and differentiation of ADSCs in fibrin scaffolds.

Cadmium Testicular Toxicity in Male Wistar Rats: Protective Roles of Zinc and Magnesium.

Cadmium (Cd) is a highly toxic element, which may cause toxicity to most organs in the body. Zinc (Zn) and magnesium (Mg) are essential minerals with probable benefits on Cd harmful effects. Finding an efficient and non-pathological treatment against Cd toxicity seems promising. Fifty adult rats were divided into ten experimental groups of five rats each. The Cd group was treated with 1 mg Cd/kg and the control group received 0.5 cm3 normal saline. The other eight groups received Zn (0.5 and 1.5 mg/kg) and Mg (0.5 and 1.5 mg/kg) either alone or in combination with 1 mg Cd/kg through IP injection for 3 weeks. Testis malondialdehyde (MDA), sperm parameters, and testis histopathology were investigated. Cd reduced sperm parameters and increased testis MDA. Moreover, Cd exposure caused a significant histological damage in testis of male rats. However, Zn or Mg treatment prevented and reversed Cd toxic alterations in testis. These findings suggest that co-administration of Zn or Mg could improve cadmium testicular toxicity in male Wistar rats.

Femoral articular geometry and patellofemoral stability.

BACKGROUND: Patellofemoral instability is a major cause of anterior knee pain. The aim of this study was to examine how the medial and lateral stability of the patellofemoral joint in the normal knee changes with knee flexion and measure its relationship to differences in femoral trochlear geometry. METHODS: Twelve fresh-frozen cadaveric knees were used. Five components of the quadriceps and the iliotibial band were loaded physiologically with 175N and 30N, respectively. The force required to displace the patella 10mm laterally and medially at 0°, 20°, 30°, 60° and 90° knee flexion was measured. Patellofemoral contact points at these knee flexion angles were marked. The trochlea cartilage geometry at these flexion angles was visualized by Computed Tomography imaging of the femora in air with no overlying tissue. The sulcus, medial and lateral facet angles were measured. The facet angles were measured relative to the posterior condylar datum. RESULTS: The lateral facet slope decreased progressively with flexion from 23°±3° (mean±S.D.) at 0° to 17±5° at 90°. While the medial facet angle increased progressively from 8°±8° to 36°±9° between 0° and 90°. Patellar lateral stability varied from 96±22N at 0°, to 77±23N at 20°, then to 101±27N at 90° knee flexion. Medial stability varied from 74±20N at 0° to 170±21N at 90°. There were significant correlations between the sulcus angle and the medial facet angle with medial stability (r=0.78, p<0.0001). CONCLUSIONS: These results provide objective evidence relating the changes of femoral profile geometry with knee flexion to patellofemoral stability.

Thymoquinone as a natural spermostatic substance in reproductive medicine: An experimental study.

BACKGROUND: Nonoxynol-9 a nonionic surfactant is widely used for its spermicidal effects. Finding new sperm immobilizing agents is necessary because Nonoxynol-9 damages the tissues of female reproductive system. OBJECTIVE: The aim of this study was to evaluate the effects of Thymoquinone (TQ) as a potential spermostatic compound on the motility and viability of human spermatozoa. MATERIALS AND METHODS: In this experimental study, the effects of 5, 10, 20, 50, 100 µg/ml, 1 and 10 mg/ml of TQ on normozoospermic semen samples were investigated. Sperm motility and viability were compared between untreated and TQ-treated aliquots of each semen sample. To evaluate the effects of TQ on the alteration of mitochondrial membrane potential (MMP), 32 semen samples were examined using 50 µg/ml of TQ. Flow cytometric analysis was performed after staining of spermatozoa with JC-1. RESULTS: Doses above 20 µg/ml of TQ could eventually immobilize all spermatozoa in culture medium. Adding 50 µg/ml of TQ did not significantly diminish the percentage of viable spermatozoa and flow cytometry results revealed that this amount of TQ could decrease sperm MMP. CONCLUSION: TQ could discontinue the movement of sperm cells in medium without reducing the population of live spermatozoa. It is more likely that TQ exerts its spermostatic action by mitigating the MMP of spermatozoa. Therefore, TQ could be considered as a potential new natural spermostatic chemical.

Effect of Thymoquinone on P53 Gene Expression and Consequence Apoptosis in Breast Cancer Cell Line.

BACKGROUND: Nigella sativa has been a nutritional flavoring factor and natural treatment for many ailments for so many years in medical science. Earlier studies have been reported that thymoquinone (TQ), an active compound of its seed, contains anticancer properties. Previous studies have shown that TQ induces apoptosis in breast cancer cells but it is unclear the role of P53 in the apoptotic pathway. Hereby, this study reports the potency of TQ on expression of tumor suppressor gene P53 and apoptosis induction in breast cancer cell line Michigan Cancer Foundation-7 (MCF-7). METHODS: MCF-7 cell line was cultured and treated with TQ, and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay was carried out for evaluating the half-maximal inhibitory concentration (IC50) values after 24 h of treatment. The percentage of apoptotic cells was measured by flow cytometry. Real-time polymerase chain reaction (PCR) was performed to estimate the messenger RNA expression of P53 in MCF-7 cell line at different times. RESULTS: The IC50 value for the TQ in MCF-7 cells was 25 µM that determined using MTT assay. The flow cytometry and real-time PCR results showed that TQ could induce apoptosis in MCF-7 cells, and the P53 gene expression was dramatically up-regulated by ascending time, respectively. Hence, there was significant difference in 48 and 72 h. CONCLUSIONS: Our results demonstrated that TQ could induce apoptosis in MCF-7 cells through up-regulation of P53 expression in breast cancer cell line (MCF-7) by time-dependent manner.

Coadministration of calcium chloride with lead acetate can improve motility of cauda epididymal spermatozoa in Swiss white mice.

BACKGROUND: Lead is an industrial heavy metal that can decrease sperm motility. OBJECTIVE: The aim was to investigate the protective effects of calcium against lead on motility of spermatozoa. MATERIALS AND METHODS: In total 40 adult male Swiss white mice were randomly divided into 5 groups (control, lead of 1(st) wk, lead of 2(nd) wk, lead/calcium of 1(st) wk and lead/calcium of 2(nd) wk). The lead groups of mice were injected by a single dose of lead acetate (200 mg/kg) intraperitoneally. Lead/calcium groups of mice were injected by a single same dose of lead acetate along with three doses of 80 mg/kg calcium chloride. The control group of mice was injected only with same volume of distilled water through the same route. Mice of 1(st) and 2(nd) wk groups were sacrificed through cervical dislocation one and two weeks after injections respectively. RESULTS: Mean of the progressive motile spermatozoa of cauda epididymis in lead/calcium group of the first week was higher than the lead group of the first week and this difference was significant. There was not any significant difference among weight of testes and epididymides of all groups. CONCLUSION: It can be concluded that calcium can decrease the effects of lead on sperm motility.

Impact of sperm chromatin evaluation on fertilization rate in intracytoplasmic sperm injection.

BACKGROUND: Sperm DNA in human beings and most vertebrates is packed by protamines into highly compact form of chromatin. There are many staining methods to assess sperm chromatin. Three different methods of staining were used simultaneously in this study and the goal was to determine which of these sperm tests has a relation with fertilization rate in intracytoplasmic sperm injection (ICSI). MATERIALS AND METHODS: Thirty couples who referred to Yamagata University Hospital (Yamagata, Japan) for ICSI were included in this study. The greater part of semen was prepared for ICSI. The remaining part was used for staining with aniline blue, acridine orange, and chromomycin A3 (CMA3). For evaluation of abnormal morphology and abnormality of head, Papanicolaou-stained smears were used. The analysis of data was done using Spearman coefficient of correlation and logistic regression model. Receiver operator characteristic (ROC) curve was used for discrimination of CMA3 staining power to identify ICSI rates. RESULTS: Percentage of CMA3 positivity, unlike those of aniline blue and acridine orange, showed significant negative correlation with fertilization rate. Moreover, the percentage of CMA3 positivity showed a positive correlation with the percentage of abnormal morphology and abnormality of head. By dividing patients into CMA3 <48% and CMA3> 48% groups, the area under the curve was 0.646. CONCLUSIONS: CMA3 staining (protamine deficiency) could be considered as a useful tool for evaluation of male fertility prior to infertility treatment.

The effects of protamine deficiency on ultrastructure of human sperm nucleus.

BACKGROUND: Chromomycin A3 (CMA3) staining is one of the staining methods for detecting protamine deficiency in sperm nucleus. CMA3 is a fluorochrome that competes with protamines for binding to DNA double helix. It has been shown in our previous studies that percentage of CMA3 positive spermatozoa in semen has a close significant relationship with the fertilization rate in in vitro fertilization (IVF). The aim of this study was to examine the ultrastructural differences between sperms in patients who had high fluorescent percentages of yellow or red in CMA3 staining (protamine deficient) with patients with low fluorescent percentages. MATERIALS AND METHODS: Semen samples are taken from five patients with high fluorescent percentages and five patients with low fluorescent percentages. Then the samples are passed for the different steps of preparing for electron microscopy. After the sectioning and mounting on grids, they are investigated under the transmission electron microscope. RESULTS: Sperms in patients with low percentages of positive spermatozoa often have a normal appearance. Sperms in high fluorescent samples frequently have unpacked chromatin. Furthermore acrosomes of these sperms are thinner or disturbed. Also sometimes there are irregularities in sperm head membrane. CONCLUSION: Protamine deficiency in sperm nucleus can cause ultrastructural anomalies in sperm chromatin such as unpacking of it. It also is concomitant with acrosome and sperm membrane disturbances.