Exploring associations between the teat apex metagenome and Staphylococcus aureus intramammary infections in primiparous cows under organic directives.

The primary objective of this study was to identify associations between the prepartum teat apex microbiome and the presence of Staphylococcus aureus intramammary infections (IMI) in primiparous cows during the first 5 weeks after calving. We performed a case-control study using shotgun metagenomics of the teat apex and culture-based milk data collected longitudinally from 710 primiparous cows on five organic dairy farms. Cases had higher odds of having S. aureus metagenomic DNA on the teat apex prior to parturition compared to controls (OR = 38.9, 95% CI: 14.84-102.21). Differential abundance analysis confirmed this association, with cases having a 23.8 higher log fold change (LFC) in the abundance of S. aureus in their samples compared to controls. Of the most prevalent microorganisms in controls, those associated with a lower risk of post-calving S. aureus IMI included Microbacterium phage Min 1 (OR = 0.37, 95% CI: 0.25-0.53), Corynebacterium efficiens (OR = 0.53, 95% CI: 0.30-0.94), Kocuria polaris (OR = 0.54, 95% CI: 0.35-0.82), Micrococcus terreus (OR = 0.64, 95% CI: 0.44-0.93), and Dietzia alimentaria (OR = 0.45, 95% CI: 0.26-0.75). Genes encoding for Microcin B17 AMPs were the most prevalent on the teat apex of cases and controls (99.7% in both groups). The predicted abundance of genes encoding for Microcin B17 was also higher in cases compared to controls (LFC 0.26). IMPORTANCE: Intramammary infections (IMI) caused by Staphylococcus aureus remain an important problem for the dairy industry. The microbiome on the external skin of the teat apex may play a role in mitigating S. aureus IMI risk, in particular the production of antimicrobial peptides (AMPs) by commensal microbes. However, current studies of the teat apex microbiome utilize a 16S approach, which precludes the detection of genomic features such as genes that encode for AMPs. Therefore, further research using a shotgun metagenomic approach is needed to understand what role prepartum teat apex microbiome dynamics play in IMI risk.

Tuberculosis infection in HIV vs. non-HIV patients.

OBJECTIVES: Tuberculosis (TB) is the most common opportunistic infection and cause of mortality among people living with HIV, and it is possible that it may also influence the evolution of the HIV infection. We assessed the differences between HIV-positive and -negative people infected with TB. METHODS: The present study is a cross-sectional retrospective study by electronic record revision. We included patients admitted to a tertiary hospital with a diagnosis of TB between 2011 and 2016, comparing those with HIV coinfection with non-HIV patients, according to demographic and clinical characteristics. RESULTS: This study included 591 patients, of whom 32% were HIV-coinfected. HIV-TB patients were younger, with a predominance of male gender. Considering TB risk factors, there was a higher prevalence of homelessness and intravenous drug use in the HIV group. In the non-HIV group, direct contact with other patients with TB and immunosuppression were more prevalent. Relative to TB characteristics, the HIV-coinfected group presents with a higher prevalence of disseminated disease and a higher occurrence of previous TB infection. Cancer was the most frequent cause of immunosuppression in the HIV group and the number testing positive for TB via microbiological culture was lower. Assessment of microbiological resistance and in-hospital mortality showed similar numbers in both groups. CONCLUSIONS: There are few papers comparing clinical course of TB between HIV-infected and non-infected patients. Our study differs from others in the literature as we focused on a country with middling incidence of TB and further characterized the differences between HIV-infected and non-infected patients which can contribute to the management of these patients.

Pneumocystosis pneumonia: A comparison study between HIV and non-HIV immunocompromised patients.

Pneumocystis pneumonia (PCP) is caused by the fungus Pneumocystis jirovecii, and its incidence has been on the rise in immunosuppressed patients without HIV. We performed a cross sectional study in patients with PCP and assessed demographic, clinical presentation and outcome measures such as mechanical ventilation and mortality differences between HIV and non-HIV patients. The two groups were statistically significantly different, with the HIV group being younger (45.5 years vs 55.9 years, p-value 0.001) and mostly composed of male patients (69% vs 31%, p-value <0.001). Also, the HIV patients had higher percentage of respiratory complaints (90% vs 68%, p-value 0.02) and lactate dehydrogenase elevation (73% vs 40%, p-value 0.001). In contrast, non-HIV patients had worse outcomes with higher incidence of invasive mechanical ventilation (23% vs 46%, p-value 0.005) and in-hospital mortality (13% vs 37%, p-value 0.002). These results reflect the literature and should raise awareness to a potentially fatal medical situation of increasing incidence.

Skeletal maturation, fundamental motor skills, and motor performance in preschool children.

Relationships among skeletal age (SA), body size and fundamental motor skills (FMS) and motor performance were considered in 155 boys and 159 girls 3-6 years of age. Stature and body mass were measured. SA of the hand-wrist was assessed with the Tanner-Whitehouse II 20 bone method. The Test of Gross Motor Development, 2nd edition (TGMD-2), and the Preschool Test Battery were used, respectively, to assess FMS and motor performance. Based on hierarchical regression analyses, the standardized residuals of SA on chronological age (SAsr) explained a maximum of 6.1% of the variance in FMS and motor performance in boys (ΔR2 3 , range 0.0%-6.1%) and a maximum of 20.4% of the variance in girls (ΔR2 3 , range 0.0%-20.4%) over that explained by body size and interactions of SAsr with body size (step 3). The interactions of the SAsr and stature and body mass (step 2) explained a maximum of 28.3% of the variance in boys (ΔR2 2 , range 0.5%-28.3%) and 16.7% of the variance in girls (ΔR2 2 , range 0.7%-16.7%) over that explained by body size alone. With the exception of balance, relationships among SAsr and FMS or motor performance differed between boys and girls. Overall, SA per se or interacting with body size had a relatively small influence in FMS and motor performance in children 3-6 years of age.

Discovery and characterization of new microsatellite loci in Dipteryx alata Vogel (Fabaceae) using next-generation sequencing data.

The use of next-generation sequencing (NGS) technologies provides a great volume of genome sequence data even for non-model species. The development of microsatellite markers using these data is a relatively quick and easy process. Dipteryx alata Vogel (Fabaceae) is an arboreal species from the Cerrado biome and is considered an important plant genetic resource. Here, we report the development of microsatellite markers for D. alata using NGS data. DNA samples from four individuals were sequenced using the Illumina MiSeq platform and high-quality reads were assembled into contigs of the D. alata genome sequence. Microsatellite regions were identified using the IMEX webserver and primer pairs were designed using the Primer3 software. The amplification settings for each locus were optimized. Fluorescent-labeled primers were developed and used to genotype individuals derived from three natural populations of D. alata. Fifty-four microsatellite regions were identified, from which 27 were elected to primer design. Among the amplified loci, 11 were polymorphic, with the number of alleles ranging from 2 to 10. The expected heterozygosity under Hardy-Weinberg Equilibrium (HWE) per locus varied from 0.191 to 0.807. Genotype and allele frequencies for all loci agreed with those expected under HWE and linkage disequilibrium was not significant for all pairs of loci. The probabilities of exclusion of paternity and of combined identity were equal to 0.993 and 5.65 x 10-8, respectively. The markers developed in this study are useful to several types of population genetic studies with D. alata and, eventually, for closely related species.

Sex differences in hepatic and intestinal contributions to nevirapine biotransformation in rats.

The understanding of the intestine contribution to drug biotransformation improved significantly in recent years. However, the sources of inter-individual variability in intestinal drug biotransformation, namely sex-differences, are still elusive. Nevirapine (NVP) is an orally taken anti-HIV drug associated with severe idiosyncratic reactions elicited by toxic metabolites, with women at increased risk. As such, NVP is a good model to assess sex-dimorphic metabolism. The aim of this study was to perform a comparative profiling of NVP biotransformation in rat intestine and liver and evaluate whether or not it is organ- and sex-dependent. Therefore, nevirapine-containing solutions were perfused through the intestine, in a specially designed chamber, or incubated with liver slices, from male and female Wistar rats. The levels of NVP and its Phase I metabolites were quantified by HPLC-UV. Liver incubation experiments yielded the metabolites 2-, 3-, 8-, and 12-OH-NVP, being 12-OH-NVP and 2-OH-NVP the major metabolites in males and females, respectively. Inter-sex differences in the metabolic profile were also detected in the intestine perfusion experiments. Herein, the metabolites 3- and 12-OH-NVP were only found in male rats, whereas 2-OH-NVP levels were higher in females, both in extraluminal (p<0.01) and intraluminal media. The metabolite 8-OH-NVP was not detected in the intraluminal media from either males or females. In this study, important inter-sex differences were detected in both organs, providing further clues to the sex-dimorphic profile of NVP toxicity. Moreover, an extra-hepatic contribution to NVP biotransformation was observed, strengthening the relevance of the intestinal contribution in the biotransformation of orally taken-drugs.

Prophylactic use of liposomal amphotericin B in preventing fungal infections early after liver transplantation: a retrospective, single-center study.

In this study the authors evaluated the efficacy of prophylaxis with liposomal amphotericin B (L-AmB) in the incidence of fungal infections (FI) during the first 3 months after liver transplant (LT). The study was retrospective and accessed a 4-year period from 2008 to 2011. All patients who died in the first 48 hours after LT were excluded. Patients were divided by the risk groups for FI: Group 1, high-risk (at least 1 of the following conditions: urgent LT; serum creatinine >2 mg/dL; early acute kidney injury [AKI] after LT; retransplantation; surgical exploration early post-LT; transfused cellular blood components [>40 U]); and Group 2, low-risk patients. Group 1 patients were further separated into those who received antifungal prophylaxis with L-AmB and those who did not. Prophylaxis with L-AmB consisted of intravenous administration of L-AmB, 100 mg daily for 14 days. Four hundred ninety-two patients underwent LT; 31 died in the first 48 hours after LT. From the remaining 461 patients, 104 presented with high-risk factors for FI (Group 1); of these, 66 patients received antifungal prophylaxis and 38 did not. In this group 8 FI were observed, 5 in patients without antifungal prophylaxis (P = .011). Three more FI were identified in Group 2. By logistic regression analysis, the categorical variable high-risk group was independently related to the occurrence of invasive FI (P = .006). We conclude that prophylaxis with L-AmB after LT was effective in reducing the incidence of FI. No influence on mortality was detected.

N-terminal valine adduct from the anti-HIV drug abacavir in rat haemoglobin as evidence for abacavir metabolism to a reactive aldehyde in vivo.

BACKGROUND AND PURPOSE: The aim of this study was to obtain evidence for the activation of the nucleoside reverse transcriptase inhibitor abacavir to reactive aldehyde metabolites in vivo. Protein haptenation by these reactive metabolites may be a factor in abacavir-induced toxic events. EXPERIMENTAL APPROACH: The formation of N-terminal valine adducts from the abacavir-derived aldehydes was investigated in the haemoglobin of Wistar rats treated with eight daily doses (120 mg·kg(-1)) of abacavir. The analyses were conducted by high-performance liquid chromatography-electrospray ionization-tandem mass spectrometry upon comparison with synthetic standards. KEY RESULTS: An N-terminal valine haemoglobin adduct derived from an α,ß-unsaturated aldehyde metabolite of abacavir was identified in vivo for the first time. CONCLUSIONS AND IMPLICATIONS: This preliminary work on abacavir metabolism provides the first unequivocal evidence for the formation of an α,ß-unsaturated aldehyde metabolite in vivo and of its ability to form haptens with proteins. The methodology described herein can be used to assess the formation of this metabolite in human samples and has the potential to become a valuable pharmacological tool for mechanistic studies of abacavir toxicity. In fact, the simplicity of the method suggests that the abacavir adduct with the N-terminal valine of haemoglobin could be used to investigate abacavir-induced toxicity for accurate risk/benefit estimations.

Percutaneous occlusion of patent foramen ovale in patients with paradoxical embolism.

INTRODUCTION: A patent foramen ovale can be found in about one quarter of adults and in a small percentage it is a wide opening and may be associated with aneurysmal formation. The association between a wide patent foramen ovale and paradoxical embolism is well established. In such cases percutaneous closure is indicated, as an alternative to life-long anticoagulant therapy or surgery. Percutaneous closure is an attractive technique and is more advantageous than other methods. METHODOLOGY: We describe the first cases of percutaneous occlusion of patent foramen ovale performed in Portugal, using the Amplatzer PFO occluder, in three female patients with documented cerebrovascular accidents due to paradoxical embolism. We also analyze the rationale for using this technique in such patients and its preliminary results. RESULTS: All three patients submitted to percutaneous occlusion of patent foramen ovale had a similar history of ischemic cerebrovascular accident. Transesophageal echocardiography showed a wide-open foramen ovale ranging from 9 to 12 mm, with spontaneous right-to-left shunt in all patients, and one of them also had an aneurysmal formation. Total procedure time ranged from 30 to 55 minutes and fluoroscopic time from 9 to 12 minutes. There were no complications and during the short follow up all patients are asymptomatic and free of recurrent events. CONCLUSIONS: Percutaneous closure of patent foramen ovale is a safe and promising technique in the prevention of recurrent systemic thromboembolism in appropriately selected patients. Prospective studies comparing antithrombotic therapy or surgery with percutaneous closure should clarify its efficacy and therapeutic value.

[A case of Ebstein disease and its surgical repair with dura-mater prosthesis]. / Um caso de doença de Ebstein e sua correcção cirúrgica com prótese de dura-mater.

The A.A. present a case of Ebstein's Disease, on a woman 48 years old. After stressing the clinical and laboratorial aspects, as well as hemodynamics and angiocardiography data, they summarize the operation, on which was used a human "dura-mater" valve in tricuspid position. There is no knowledge on literature of previous applications of this kind of valve on Ebstein's Disease. The patient, one year after the operation, is improving quiet well.

Early and medium term results of tailored therapy for heart failure.

UNLABELLED: Patients (pts) with advanced chronic heart failure, in NYHA functional class IV, refractory to conventional medical therapy, show a poor short-term survival prognosis. Heart transplant remains the therapy of choice but it can currently be performed in only a minority of pts. Therapy tailored to hemodynamic goals has been suggested as a potential alternative for patients with advanced congestive heart failure. Intravenous and, subsequently, oral vasodilators (v) and diuretics (d) are titrated, in order to achieve specific hemodynamic parameters: systolic arterial pressure (SAP) > 80 mmHg, pulmonary wedge pressure (PWP) < 15 mmHg, right atrial pressure (RAP) < 8 mmHg, and systemic vascular resistance (SVR) < 1200 dynes.sec.cm-5. AIM: To assess short and medium term (two years) results of a tailored therapy management program for treatment of patients with advanced heart failure. METHODS: 27 pts (19 males, 61 +/- 10 years), NYHA functional class IV, with dilated cardiomyopathy (13 idiopathic, 10 ischemic, 4 hypertensive), 17 with exclusion criteria for heart transplantation, were included. Echocardiographic left ventricular end-diastolic dimension and ejection fraction were 68 +/- 8 mm and 20 +/- 9%, respectively. Initial (i) serum sodium (Na+) was 136 +/- 5 mEq/l and i serum creatinine (Cr) was 1.4 +/- 0.8 mg/dl. Baseline, at referral, hemodynamics: SAP = 125 +/- 23, PWP = 23 +/- 6, RAP = 12 +/- 6, cardiac index (CI) = 1.9 +/- 0.5 l/min/m2, SVR = 2193 +/- 670. Using bedside right heart catheterization (Swan-Ganz catheter) we set out to achieve the above hemodynamic goals. RESULTS: 1) v and d used: sodium nitroprusside--cumulative dose = 196 +/- 121 mg, captopril--daily dose (dd) = 157 +/- 95 mg, isosorbide dinitrate--dd = 91 +/- 57 mg, hydralazine--dd = 95 +/- 67 mg, and furosemide--dd = 105 +/- 70 mg; 2) final (f) hemodynamic parameters on tailored therapy: SAP = 109 +/- 20*, PWP = 12 +/- 3*, RAP = 4 +/- 3*, CI = 2.5 +/- 0.6*, SVR = 1317 +/- 340* (*: p < 0.001 vs baseline); 3) duration of invasive monitoring was 3.0 +/- 1.9 days; 4) f Na+ = 134 +/- 5, and f Cr = 1.5 +/- 0.8 (NS vs i); 5) there was one (4%) in-hospital death; functional class of discharged pts: III--4 pts, II--18 pts, I--4 pts; 6) nine pts (35%) died after discharge--three due to refractory heart failure and six (including two potential heart transplant candidates) had sudden death; 7) actuarial survival (Kaplan-Meyer method): at 6 months (m)--80%, 12 m--71%, 18 m--64%, 24 m--55%; 8) after a mean follow-up of 18 +/- 8 m, functional class in survivors was: NYHA IV--2 pts, III--4 pts, II--8 pts, and I--3 pts. CONCLUSIONS: Therapy tailored to hemodynamic goals is a valid approach for pts with advanced heart failure, showing good hemodynamic and functional short-term results, and reasonable survival at two years. The significant incidence of sudden death demands strategies for risk stratification and a search for prophylactic measures in this population.

Non-invasive monitoring of pulmonary capillary wedge pressure in heart failure.

OBJECTIVE: To evaluate the usefulness of pulsed Doppler (PD) mitral flow E wave deceleration time (EDcT) to detect and quantify changes of pulmonary capillary wedge pressure (PCWP) in patients (pt) with dilated chronic heart failure (CHF) submitted to tailored therapy. METHODS: In 14 pt with dilated cardiomyopathy (DCM) (59.4+/-10.0 years, 11 males, sinus rhythm), admitted to the ICU because of worsening CHF, serial simultaneous hemodynamic and echocardiographic studies were performed (3-5/pt; overall 49 evaluations). PD mitral flow register was used to measure EDcT and correlated with PCWP at each study. RESULTS: PCWP ranged from 36 to 3 mmHg (17.6+/-8.8) and EDcT from 271 to 52 ms (104.9+/-42.4). The correlation between EDcT and PCWP was -0.65 (PCWP=31.7-0.134EDcT). Using this equation to calculate PCWP, individual absolute values difference (identity error - IE) was 5.6+/-3.5 mmHg (0.4-14.6). In 29 cases (59.2%) IE was >5 mmHg, defined as major error. Considering EDcT percent change (delta%EDcT) and PCWP variation (deltaPCWP) in serial evaluations, we found a correlation of -0.87 (deltaPCWP=-2.83-0.19 delta%EDcT). Using this equation, the IE was 2.3+/-1.6 mmHg (0-5.2) and there were only two (6%) major errors (P<0.0001). CONCLUSIONS: In pt with DCM and advanced CHF, EDcT shows a reasonable correlation with PCWP, but when it is used to calculate PCWP the IE to hemodynamic values is often large. However, our results with EDcT percent change in serial evaluations using a first simultaneous invasive determination, suggest that this technique is reliable for monitoring PCWP and can be particularly useful for pt submitted to tailored therapy.

New criteria for the diagnosis and treatment of atrioventricular node reentrant tachycardia.

The authors describe a new elecrocardiographic criterion to diagnose atrioventricular node reentrant tachycardia, review the electrocardiographic and electrophysiological diagnosis of this tachyarrhythmia and analyse the results of catheter ablation of fast versus slow pathway, selecting the fast pathway whenever the induction of atrioventricular node reentrant tachycardia is performed only after pharmacological maneuvers. The high specificity for atrioventricular node reentrant tachycardia of the criterion characterized by the absence of ST-segment depression found in left precordial leads from V4 to V6 suggests the need to include it in electrocardiographic algorithms for the differential diagnosis of tachycardias with a narrow QRS complex. The high primary and final success rates of catheter ablation obtained after a therapy stratification based on induction criteria, emphasize the use of the fast pathway ablation as a first option in selected cases and for the treatment of recurrences.

[Diagnosis of paradoxal and concomitant pulmonary thromboembolism with transesophageal echocardiography: a clinical case]. / Diagnóstico de tromboembolismo paradoxal e pulmonar concomitante por ecocardiografia transesofágica: um caso clínico.

There is some controversy regarding the indications for transesophageal echocardiography in patients with suspected systemic embolism. The present case report refers to a 65 year old male admitted to the hospital for ischemic acute cerebrovascular accident, which was confirmed by cerebral computerized tomography. A transthoracic echocardiogram was performed showing right atrial and ventricular dilatation. A transesophageal echocardiogram was also performed to exclude thromboembolism and clarify dilatation of the right cavities. Mild spontaneous echocontrast was present in the left atrium without images of thrombus; an interatrial septal aneurysm with patent foramen ovale was found with right to left flow; an image compatible with a very mobile, large, proximal thrombus in the main pulmonary artery was observed. A venous duplex scan was performed, demonstrating venous thrombosis in the right popliteal and femoral veins. Pulmonary arteriography showed a large thrombus in the right pulmonary branch extending to the median lobe and a smaller thrombus in the left pulmonary branch. Apparently, the patient had no predisposing factors for thromboembolism. Full-dose heparin was started followed by oral anticoagulation. An inferior vena cava filter was implanted. At hospital discharge the pulmonary thrombus had disappeared and the right cardiac cavities had returned to normal size. The interatrial aneurysm had disappeared and foramen ovale was no longer patent. After 36 months of clinical follow up on oral anticoagulation, the patient remains asymptomatic without neurological sequelae nor respiratory distress.

Nonpharmacological treatment of atrial tachycardia.

The authors analyse the efficacy and safety of catheter ablation and atrial pacing for the treatment of atrial tachycardia. Radiofrequency catheter ablation was selected whenever the arrhythmogenic focus was located on the free-wall or in the meso-septal area of the right atrium. In opposition, overdrive atrial pacing was chosen for tachycardias originating near the sinus complex or in the left atrium. Both therapies were safe, but had a low efficacy in converting the tachycardia into sinus rhythm. However, catheter ablation allows an irreversible destruction of small septally located foci. Thus, both the anatomical and the electrophysiological characteristics of the foci can be important factors in the selection of the most appropriate nonpharmacologic therapy.

Cardiac output quantification by Doppler echocardiography in intensive care--limitations and validation.

UNLABELLED: Cardiac output (CO) quantification is primordial to the evaluation of patients with heart failure who are on tailored therapy and under invasive hemodynamic monitoring. Doppler echocardiography can be used to access CO noninvasively, but the concordance between its results and those obtained by invasive methods in paired measurements is still controversial. To our knowledge, no previous studies have assessed the clinical relevance of Doppler echocardiography for CO serial evaluation in patients submitted to tailored therapy. AIM: To evaluate the usefulness of echo-Doppler in the assessment of CO and quantification of changes in CO, compared to thermodilution, in patients with advanced heart failure under hemodynamic monitoring to guide tailored therapy. METHODS: In 20 patients (14 male, 62 +/- 14 years old, all in sinus rhythm), with dilated cardiomyopathy and NYHA IV, admitted to the intensive care unit (ICU), CO was simultaneously determined by Doppler echocardiography (dpCO) and thermodilution (tdCO) in three serial evaluations (overall 60). The dpCO was calculated by multiplying the aortic orifice area by the velocity-time integral of aortic continuous wave Doppler flow and by the heart rate. A difference between tdCO and dpCO of more than 20% was considered a major error. RESULTS: In the overall evaluations, dpCO systematically overestimates tdCO (p = 0.026). The correlation between tdCO and dpCO was 0.81, the mean difference between measurements was 0.40 +/- 0.61 l/min (mean -2SD = -1.62 mean +2SD = 0.81) and 19 (32%) major errors occurred. No significant difference was found between CO percentual variation values assessed by both methods, with a stronger correlation (r = 0.92-p = 0.014) compared to that found for absolute values. On using the dpCO/tdCO ratio in the first evaluation to correct subsequent dpCO, the correlation was fairly good (r = 0.96-p = 0.0002 versus corrected dpCO). The mean difference between paired measurements was significantly lower (0.12 +/- 0.28 l/min-mean 2SD = -0.44 mean +2SD = 0.67), and there were no errors. CONCLUSIONS: CO estimated by Doppler echocardiography has a good correlation with thermodilution although with a weak concordance between paired results in patients with dilated cardiomyopathy and advanced heart failure admitted to the ICU for tailored therapy. Our results with dpCO percentual change in repeated evaluations and with corrected dpCO value after a single simultaneous invasive determination suggest that Doppler echocardiography is a valid method for clinical purposes, allowing us to propose a reduction in the time period of invasive hemodynamic monitoring.