Objective evaluation of ram and buck sperm motility by using a novel sperm tracker software.

This work offers researchers the first version of an open-source sperm tracker software (Sperm Motility Tracker, V1.0) containing a novel suit of algorithms to analyze sperm motility using ram and buck sperm as models. The computer-assisted semen analysis is used in several publications with increasing trend worldwide in the last years, showing the importance of objective methodologies to evaluate semen quality. However, commercial systems are costly and versatility is constrained. In the proposed method, segmentation is applied and the tracking stage is performed by using individual Kalman filters and a simplified occlusion handling method. The tracking performance in terms of precision (number of true tracks), the percentage of fragmented paths and percentage of correctly detected particles were manually validated by three experts and compared with the performance of a commercial motility analyzer (Microptic's SCA). The precision obtained with our sperm motility tracker was higher than the one obtained with a commercial software at the current acquisition frame rate of 25 fps (P < 0.0001), concomitantly with a similar percentage of fragmentized tracks (P = 0.0709) at sperm concentrations ranging 25-37 × 106 cells/mL. Moreover, our tracker was able to detect trajectories that were unseen by SCA. Kinetic values obtained by using both methods were contrasted. The higher values found were explained based on the better performance of our sperm tracker to report speed parameters for very fast motile sperm. To standardize results, acquisition conditions are suggested. This open-source sperm tracker software has a good plasticity allowing researchers to upgrade according requirements and to apply the tool for sperm from a variety of species.

SDF-1 in myocardial repair.

Stem cell therapy for the prevention and treatment of cardiac dysfunction holds significant promise for patients with ischemic heart disease. Excitingly early clinical studies have demonstrated safety and some clinical feasibility, while at the same time studies in the laboratory have investigated mechanisms of action and strategies to optimize the effects of regenerative cardiac therapies. One of the key pathways that has been demonstrated critical in stem cell-based cardiac repair is (stromal cell-derived factor-1) SDF-1:CXCR4. SDF-1:CXCR4 has been shown to affect stem cell homing, cardiac myocyte survival and ventricular remodeling in animal studies of acute myocardial infarction and chronic heart failure. Recently released clinical data suggest that SDF-1 alone is sufficient to induce cardiac repair. Most importantly, studies like those on the SDF-1:CXCR4 axis have suggested mechanisms critical for cardiac regenerative therapies that if clinical investigators continue to ignore will result in poorly designed studies that will continue to yield negative results.

Objective and subjective sleep measures are associated with HbA1c and insulin sensitivity in the general population: Findings from the ORISCAV-LUX-2 study.

AIM: To analyze the association of objective and subjective sleep measures with HbA1c and insulin sensitivity in the general population. METHODS: Using a cross-sectional design, data from 1028 participants in the ORISCAV-LUX-2 study from the general population in Luxembourg were analyzed. Objective sleep measures were assessed using accelerometers whereas subjective measures were assessed using the Pittsburgh Sleep Quality Index (PSQI) questionnaire. Sleep measures were defined as predictors, while HbA1c and quantitative insulin sensitivity check index (QUICKI) scores were considered outcomes. Linear and spline regression models were fitted by progressively adjusting for demographic and lifestyle variables in the total sample population as well as by stratified analyses using gender, obesity status, depressive symptoms and diabetes status. RESULTS: In fully adjusted models, total and deep sleep durations were associated with lower HbA1c (mmol/mol) levels, whereas sleep coefficients of variation (%) and poor sleep efficiency, as measured by PSQI scores (units), were associated with higher HbA1c levels. In stratified models, such associations were observed mainly in men, and in subjects who had depressive symptoms, were overweight and no diabetes. In addition, total sleep, deep sleep, coefficients of variation and poor sleep efficiency as measured by PSQI revealed non-linear associations. Similarly, greater insulin sensitivity was associated with longer total sleep time and with PSQI-6 (use of sleep medication). CONCLUSION: Associations were more frequently observed between sleep characteristics and glycaemic control with the use of objective sleep measures. Also, such associations varied within subgroups of the population. Our results highlight the relevance of measuring sleep patterns as key factors in the prevention of diabetes.

Plasmid-based transient human stromal cell-derived factor-1 gene transfer improves cardiac function in chronic heart failure.

We previously demonstrated that transient stromal cell-derived factor-1 alpha (SDF-1) improved cardiac function when delivered via cell therapy in ischemic cardiomyopathy at a time remote from acute myocardial infarction (MI) rats. We hypothesized that non-viral gene transfer of naked plasmid DNA-expressing hSDF-1 could similarly improve cardiac function. To optimize plasmid delivery, we tested SDF-1 and luciferase plasmids driven by the cytomegalovirus (CMV) promoter with (pCMVe) or without (pCMV) translational enhancers or α myosin heavy chain (pMHC) promoter in a rodent model of heart failure. In vivo expression of pCMVe was 10-fold greater than pCMV and pMHC expression and continued over 30 days. We directly injected rat hearts with SDF-1 plasmid 1 month after MI and assessed heart function. At 4 weeks after plasmid injection, we observed a 35.97 and 32.65% decline in fractional shortening (FS) in control (saline) animals and pMHC-hSDF1 animals, respectively, which was sustained to 8 weeks. In contrast, we observed a significant 24.97% increase in animals injected with the pCMVe-hSDF1 vector. Immunohistochemistry of cardiac tissue revealed a significant increase in vessel density in the hSDF-1-treated animals compared with control animals. Increasing SDF-1 expression promoted angiogenesis and improved cardiac function in rats with ischemic heart failure along with evidence of scar remodeling with a trend toward decreased myocardial fibrosis. These data demonstrate that stand-alone non-viral hSDF-1 gene transfer is a strategy for improving cardiac function in ischemic cardiomyopathy.

Enterobacteria associated with houseflies (Musca domestica) as an infection risk indicator in swine production farms.

Houseflies (Musca domestica) spend part of their life development on animal or human manure. Manure is high in pathogenic microbes; thus, houseflies have been known as a mechanical vector for various important zoonotic diseases. Therefore, the present study showcases captured houseflies from intensive swine production regions (which are areas of high manure concentration) in Southern Brazil, and analyses their bodies' to the presence of Escherichia coli and Salmonella sp. and the sensitivity of these bacteria to various antibiotics. Additionally, Quantitative Microbiology Risk Assessment was performed simulating the contamination of lettuce by flies' bacteria and subsequent lettuce consumption by an adult human being. Houseflies were captured in swine buildings and farm houses from five farms. E. coli quantification values ranged from 104 to 106 CFU/20 flies, and all sampling sites had positive results from bacteria presence in the collected houseflies. On the other hand, Salmonella sp. presence was observed in only three farms, where the quantification ranged from 102 to 105 CFU/20 flies. The bacteria showed to be resistant to at least two from the four tested antibiotics (ampicillin, Cefalotin, Ciprofloxacin and Norfloxacin) antibiotics used in human or veterinary medicine. Infection probability analyses showed risk of human infection by E.coli, indicating possible transmission of zoonotic pathogens through flies. In this context, it was possible to conclude that there is a need for flies control, especially in swine farms where zoonotic pathogens can be abundant, to minimize the health impact of the vectorization of enteric bacteria.

Role of structural barriers in the mechanism of alternans-induced reentry.

Previously, using an animal model of T-wave alternans in structurally normal myocardium, we demonstrated that repolarization can alternate with opposite phase between neighboring myocytes (ie, discordant alternans), causing spatial dispersions of repolarization that form the substrate for functional block and reentrant ventricular fibrillation (VF). However, the mechanisms responsible for cellular discordant alternans and its electrocardiographic manifestation (ie, T-wave alternans) in patients with structural heart disease are unknown. We hypothesize that electrotonic uncoupling between neighboring regions of cells by a structural barrier (SB) is a mechanism for discordant alternans. Using voltage-sensitive dyes, ventricular action potentials were recorded from 26 Langendorff-perfused guinea pig hearts in the absence (ie, control) and presence of an insulating SB produced by an epicardial laser lesion. Quantitative analysis of magnitude and phase of cellular alternans revealed that in controls, action potential duration alternated in phase at all ventricular sites above a critical heart rate (269+/-17 bpm), ie, concordant alternans. Also, above a faster critical heart rate threshold (335+/-24 bpm), action potential duration alternated with opposite phase between sites, ie, discordant alternans. In contrast, only discordant but not concordant alternans was observed in 80% of hearts with the SB, and discordant alternans always occurred at a significantly slower heart rate (by 68+/-28 bpm) compared with controls. Therefore, the SB had a major effect on the alternans-heart rate relation, which served to facilitate the development of discordant alternans. Whether a SB was present or not, discordant alternans produced considerable increases (by approximately 170%) in the maximum spatial gradient of repolarization, which in turn formed the substrate for unidirectional block and reentry. However, by providing a structural anchor for stable reentry, discordant alternans in the presence of a SB led most often to sustained monomorphic ventricular tachycardia rather than to VF, whereas in the absence of a SB discordant alternans caused VF. SBs facilitate development of discordant alternans between cells with different ionic properties by electrotonically uncoupling neighboring regions of myocardium. This may explain why arrhythmia-prone patients with structural heart disease exhibit T-wave alternans at lower heart rates. These data also suggest a singular mechanism by which T-wave alternans forms a substrate for initiation of both VF and sustained monomorphic ventricular tachycardia.

Catheter-based intravascular ultrasound discriminates bicuspid from tricuspid valves in adults with calcific aortic stenosis.

A catheter-based intravascular ultrasound transducer was used to study aortic valve morphology in adults with calcific aortic stenosis. Examination of 14 postmortem specimens disclosed that intravascular ultrasound consistently identified the number of cusps or the presence of a calcified median raphe in the conjoined cusp, or both, and thereby distinguished a calcified bicuspid from a calcified tricuspid aortic valve. These postmortem findings were then employed to identify valvular morphology in 15 patients undergoing diagnostic cardiac catheterization or balloon aortic valvuloplasty, or both. Reproduction of criteria established in vitro allowed discrimination of congenital valvular morphology in all 15 patients, including 7 in whom assessment by intravascular ultrasound was confirmed by subsequent pathologic examination. Identification of aortic valvular morphology by intravascular ultrasound has potential therapeutic implications for patients considered to be candidates for operative or nonoperative aortic valvuloplasty.

Clinical issues related to discontinuing digoxin therapy in elderly nursing home patients.

Digoxin is commonly used to treat congestive heart failure. Digoxin augments ventricular systolic performance, but does not benefit patients whose congestive heart failure is caused by poor diastolic function. We studied 47 elderly nursing home patients who were receiving long-term digoxin therapy. The left ventricular ejection fractions were measured using both a standard and a highly portable echocardiography machine. Thirty-five of 47 patients had normal ejection fractions (50% or greater). In this subgroup, 23 patients were in normal sinus rhythm. Digoxin was discontinued in 14 patients with good systolic function and normal sinus rhythm, but in nine cases physicians refused to stop the digoxin. Follow-up evaluations showed no deterioration off digoxin. Excellent correlations existed between estimated left ventricular ejection fractions from the two echocardiography machines. Many nursing home patients taking digoxin do not need it. Physician reluctance to discontinue digoxin may change with the availability of highly portable echocardiography.

Intraoperative multiplane transesophageal echocardiography for guiding direct myocardial gene transfer of vascular endothelial growth factor in patients with refractory angina pectoris.

Gene transfer for therapeutic angiogenesis represents a novel treatment for patients with chronic angina refractory to standard medical therapy and not amenable to conventional revascularization. We sought to assess the role of intraoperative multiplane transesophageal echocardiography (MPTEE) in guiding injection of naked DNA encoding vascular endothelial growth factor (VEGF) into the left ventricular (LV) myocardium of patients with refractory angina. After exposing the LV myocardium via a limited lateral thoracotomy, each of 17 patients in this series received 4 separate injections of VEGF DNA into different myocardial sites. Initial injections in the first patient produced intracavitary microbubbles, indicating injection of DNA into the LV chamber. Subsequently, each injection was preceded by a test injection of agitated saline. The absence of microbubbles while visualizing the LV cavity during the test injection verified that the ensuing injection of DNA would not be inadvertently squandered in the LV chamber itself. Intracavitary LV microbubbles were observed by MPTEE in 13 of 64 (20.3%) saline test injections and in 8 of 16 (50.0%) patients in which saline test injection was used, leading to adjustments in needle position. MPTEE imaging detected a previously unknown large, apical left ventricular thrombus in one patient, thereby preventing inadvertent injection of VEGF DNA through the myocardium into the thrombus. Imaging during and after injection verified no deleterious impact on LV function. We conclude that MPTEE is a useful tool for ensuring that myocardial gene therapy performed by direct needle injection results in gene transfer to the LV myocardium.

Flavonoid inhibition of platelet procoagulant activity and phosphoinositide synthesis.

Dietary flavonoids are known for their antiplatelet activity resulting in cardiovascular protection. Phosphatidylinositol 4,5-bisphosphate (PIP2) was previously reported to play a direct role in phosphatidylserine (PS) exposure, as a Ca2+ target. Thrombin formation and platelet procoagulant activity are dependent on PS exposure. As flavonoids can inhibit phosphoinositide (PPI) kinases, we examined whether changes in PPI metabolism in flavonoid-treated platelets could be involved in their antiplatelet effects. Treatment with the flavonoids quercetin or catechin reduced PS exposure, thrombin formation, PIP2 level and resynthesis after platelet activation with collagen, thrombin or calcium ionophore. Flavonoids also prevented [Ca2+]i increase induced by collagen, but not by the ionophore. The ability of flavonoids to decrease PS exposure induced by ionophore treatment could result from the diminution of PIP2 levels, whereas PS exposure induced by collagen could also be diminished by flavonoids' effects on calcium signaling dependent on PIP2 hydrolysis. These data favor a role for PIP2 in the antiplatelet effects of flavonoids.

Cellular distribution of pulmonary Mn and CuZn superoxide dismutase: effect of hyperoxia and interleukin-1.

Pulmonary superoxide dismutase (SOD) plays an important role in the lung defense against O2 toxicity. We have previously demonstrated that tracheal insufflation of interleukin-1 alpha (IL-1) selectively enhances pulmonary MnSOD and protects rats against O2 toxicity. However, little is known about the cellular distribution of pulmonary MnSOD- and CuZnSOD-specific proteins. We performed immunohistochemistry in plastic sections (2 microns thick) to determine the effects of hyperoxia and IL-1 on the cellular distribution of pulmonary MnSOD and CuZnSOD in rats. MnSOD and CuZnSOD were present in all lung cells. Smooth muscle and endothelial cells appeared to contain higher immunoreactive MnSOD and CuZnSOD proteins than other lung cell types. Exposure of rats to 100% O2 for 24 hr had no effect on the cellular distribution and intensity of pulmonary MnSOD. However, at 50 hr after O2 exposure the intensity of pulmonary MnSOD was reduced. In contrast, tracheal insufflation of IL-1 markedly enhanced the intensity of pulmonary MnSOD in rats exposed to O2 for 50 hr. Neither O2 exposure nor IL-1 insufflation had any apparent effect on the distribution and intensity of pulmonary CuZnSOD. We conclude that IL-1 selectively enhances pulmonary MnSOD and that this effect is manifested in most lung cells, particularly smooth muscle and endothelial cells.

Effect of local lidocaine anesthesia on ventricular escape intervals during permanent pacemaker implantation in patients with complete heart block.

Transient asystole is often noted during the course of permanent pacemaker implantation in patients with complete heart block. Since subcutaneous lidocaine is frequently used as the local anesthetic agent for permanent pacemaker implantation, the effect of this drug on ventricular escape intervals was studied. Ventricular escape intervals after transient cessation of pacing were studied in 9 patients with complete heart block before and 10, 30, and 45 minutes after subcutaneous lidocaine administration for permanent pacemaker implantation. The total lidocaine dose ranged from 170 to 400 mg (1.9 to 9.5 mg/kg of body weight). Therapeutic blood levels were achieved in 7 patients. The mean ventricular escape interval before lidocaine was 1.83 +/- 0.32 seconds, which increased to 2.58 +/- 1.35, 2.96 +/- 1.06, and 2.68 +/- 1.27 seconds at 10, 30, and 45 minutes after lidocaine (p less than 0.02). The mean maximal escape interval before lidocaine was 2.06 +/- 0.30 seconds, which increased to 3.80 +/- 1.44 seconds (p less than 0.01), a mean increase of 84%. The percent increase in maximal escape interval was related directly to the peak lidocaine level achieved. After lidocaine administration, 5 patients had asystole greater than 4 seconds and 1 required resumption of pacing. Thus, subcutaneous lidocaine contributes to the occurrence of asystole seen during permanent pacemaker implantation. It is advisable to limit the amount of lidocaine administered during permanent pacemaker implantation to the minimum necessary to achieve adequate local anesthesia. Strong consideration should be given to the use of a temporary pacemaker in patients with complete heart block during permanent pacemaker implantation even in the absence of previous asystole.

Right ventricular infarction. Clinical diagnosis and differentiation from cardiac tamponade and pericardial constriction.

Twelve patients with a clinical diagnosis of right ventricular infarction are described. All had acute inferior wall myocardial infarction associated with the bedside findings of jugular venous distension, clear lungs on auscultation, and arterial hypotension. Hemodynamically, there was elevation of right-sided filling pressures not explained by normal or minimally elevated pulmonary wedge pressures. Four patients had an incorrect diagnosis of acute cardiac tamponade. However, a review of the data showed that the hemodynamic features of right ventricular infarction more closely resemble those of pericardial constriction, a point that may be helpful in distinguishing right ventricular infarction from cardiac tamponade. Invasive and noninvasive techniques that exclude the presence of pericardial fluid and suggest enlargement and abnormal contractility of the right ventricle were helpful in establishing the diagnosis of right ventricular infarction in several patients.

Cardiac surgery report cards: comprehensive review and statistical critique.

Public report cards and confidential, collaborative peer education represent distinctly different approaches to cardiac surgery quality assessment and improvement. This review discusses the controversies regarding their methodology and relative effectiveness. Report cards have been the more commonly used approach, typically as a result of state legislation. They are based on the presumption that publication of outcomes effectively motivates providers, and that market forces will reward higher quality. Numerous studies have challenged the validity of these hypotheses. Furthermore, although states with report cards have reported significant decreases in risk-adjusted mortality, it is unclear whether this improvement resulted from public disclosure or, rather, from the development of internal quality programs by hospitals. An additional confounding factor is the nationwide decline in heart surgery mortality, including states without quality monitoring. Finally, report cards may engender negative behaviors such as high-risk case avoidance and "gaming" of the reporting system, especially if individual surgeon results are published. The alternative approach, continuous quality improvement, may provide an opportunity to enhance performance and reduce interprovider variability while avoiding the unintended negative consequences of report cards. This collaborative method, which uses exchange visits between programs and determination of best practice, has been highly effective in northern New England and in the Veterans Affairs Administration. However, despite their potential advantages, quality programs based solely on confidential continuous quality improvement do not address the issue of public accountability. For this reason, some states may continue to mandate report cards. In such instances, it is imperative that appropriate statistical techniques and report formats are used, and that professional organizations simultaneously implement continuous quality improvement programs. The statistical methodology underlying current report cards is flawed, and does not justify the degree of accuracy presented to the public. All existing risk-adjustment methods have substantial inherent imprecision, and this is compounded when the results of such patient-level models are aggregated and used inappropriately to assess provider performance. Specific problems include sample size differences, clustering of observations, multiple comparisons, and failure to account for the random component of interprovider variability. We advocate the use of hierarchical or multilevel statistical models to address these concerns, as well as report formats that emphasize the statistical uncertainty of the results.

p53 protein expression in benign lesions of the upper respiratory tract.

OBJECTIVE: p53 is a tumor suppressor gene that is lost or mutated in most forms of human malignancy. There are, however, very few studies evaluating p53 expression in normal epithelium or benign lesions. DESIGN: We screened for p53 protein expression in a variety of benign epithelial lesions of upper respiratory tract using monoclonal antibody DO-1 on paraffin-embedded material. SUBJECTS: We studied a total of 109 cases: 16 cases of juvenile and 36 cases of adult laryngeal papillomatosis, 10 cases each of laryngeal nodules and laryngeal polyps, 17 cases of inverted papilloma, and 20 cases of nasal polyps. RESULTS: Nuclear immunoreactivity for p53 protein was demonstrated in 14 (88%) of 16 cases of juvenile laryngeal papillomatosis, 33 (92%) of 36 cases of adult laryngeal papillomatosis, 4 (40%) of 10 cases of laryngeal nodules, 8 (80%) of 10 cases of laryngeal polyps, 7 (41%) of 17 cases of inverted papilloma, and 2 (10%) of 20 cases of nasal polyps. These results pertained only to the basal epithelial layer in all cases of laryngeal nodules, laryngeal polyps, and nasal polyps. Intermediate layer cells were also positive for p53 in the majority of the cases of both juvenile (69%) and adult (75%) laryngeal papillomatosis and in a minority of the cases of inverted papilloma (18%). CONCLUSIONS: Overexpression of p53 protein is commonly demonstrable in benign epithelial lesions of the upper respiratory tract. This observation suggests that p53 protein accumulation may occur in the absence of mutation of the p53 gene and may correlate with epithelial proliferative activity.

Training resident anesthesiologists in adult challenging intubation comparing Truview EVO2 and Macintosh laryngoscope: a preliminary study.

AIM: Teaching airway management continues to be of high importance to the anesthesiologist, since the care of each individual patient depends on the expertise, training and knowledge of the anesthetist with different airway devices, techniques and algorithms. The aim of our study was to compare intubation performed by resident anesthesiologists in training, under senior supervision, using Truview EVO2 (Group 1) or Macintosh blade (Group 2) in a group of adult patients undergoing elective surgery. METHODS: This was a pilot prospective study. Thirty patients who were scheduled for surgery under general anesthesia were randomized into two groups. In Group 1, intubation was performed by using the Truview EVO2, and in Group 2 intubation was performed by using the Macintosh blade. Mallampati score, thyromental distance and neck mobility were recorded for each patient. The exclusion criteria included a Mallampati score =or<2 and a Patil distance >6 cm. The time of intubation and any occurrence of complications were recorded. RESULTS: Intubation was always successful on the first attempt in Group 1, while it failed for 46.7% of patients in Group 2 (P=0.006). The time of intubation was not different between the two groups. No complications were recorded for Group 1 (Truview), while seven were reported in Group 2 (Macintosh) (P=0.003). CONCLUSIONS: The resident managed to intubate all patients on the first attempt with the Truview, which led to a lower incidence of complications. Despite the exiguity of the population in the study, Truview EVO2 and other videolaryngoscopes can be considered to be useful tools in training resident anesthesiologists in elective intubation.

A medical prescription for survival.

KIE: Lown and Pastore argue that physicians have historically fought against health hazards even when this entailed political activity. Physicians must likewise respond to the threat of nuclear warfare by supporting a comprehensive moratorium on all nuclear explosions. The International Physicians for the Prevention of Nuclear War has called for such a moratorium, and it is now scientifically possible to verify all or almost all underground explosions. The experience of the Limited Test Ban Treaty of 1963 provides a historical precedent, the authors contend, and physicians have the responsibility to work for an end to all nuclear tests.^ieng

Limitations of color flow Doppler imaging in the quantification of valvular regurgitation: velocity of regurgitant jet, rather than volume, determines size of color Doppler image.

The objective of this study was to determine the validity of estimation of regurgitant volume by visual assessment of color flow Doppler display. An experimental apparatus was designed that is capable of ejecting precise volumes of echogenic material from one chamber to another under continuous color flow Doppler monitoring. The velocity of flow was altered independently by changing either the size of the orifice through which flow occurred or the ejection rate. In this manner the differential effects of volume and velocity on the color flow Doppler image could be examined. The maximum area encompassed by the color flow Doppler pattern for each ejection was planimetered by using commercially available on-line software. In addition the reviewer in each case applied a subjective grade to the appearance of the color flow jet (1+ to 4+). Comparison was then made of the color flow Doppler appearance of equal volumes flowing at different velocities and of different volumes flowing at different velocities. In the initial series a solution of agitated hetasarch was used. When equal volumes were imaged at different velocities the higher-velocity jet appeared larger, both subjectively (3+ vs 1+) and by measuring the area encompassed in the Doppler flow profile (40.3 +/- 1.8 vs 22.0 +/- 1.4 cm2, p = 0.0001). Furthermore, when different volumes were imaged at different velocities, the smaller volume (3 ml vs 6 ml) appeared larger when it was flowing at higher velocity (3+ vs 2+, 40.3 +/- 1.8 vs 32.4 +/- 1.3 cm2, p = 0.0006). These experiments were repeated with blood, confirming the results of the initial study.(ABSTRACT TRUNCATED AT 250 WORDS)