A discrepant presentation of bacteremia in the emergency department linked to a Fusobacterium nucleatum infection: a case report

Background Fusobacterium nucleatum is an anaerobic bacterium mainly responsible for acute or chronic infection of the ear, nose, and throat, potentially bacteremic with a risk of extraoral metastatic infection. Bacteremia occurs mainly in the elderly or in immunodeficient individuals, with high mortality. F. nucleatum is not the first cause of tonsillar infection in emergency departments, which are more often the consequence of a viral or streptococcal infection, but it is a risk factor for severe bacterial infection, especially in a viral pandemic context. Case presentation A 25-year-old European woman with no history presented to the emergency department with fever (38.9 °C), pharyngeal symptoms, intermittent headaches, and alteration of general condition. On examination, she presented odynophagia associated with moderate tonsillar hypertrophy, her neck was painful but flexible. A rapid diagnostic test for beta-hemolytic group streptococcus was negative. First biological analyses revealed an inflammatory syndrome with C-reactive protein of 76 mg/L. Procalcitonin was measured secondarily, and was 2.16 µg/L. Faced with discordant clinical and biological findings, a lumbar puncture was performed, which came back negative. At hour eight, hypotension was observed but corrected after filling with physiological serum. The patient was hospitalized for monitoring, based on a hypothesis of severe viral presentation. At hour 24, pyrexia confirmed this hypothesis. A spontaneous but transient improvement and no new hemodynamic event led to early discharge. At day three, she was rehospitalized for increased and continuous headaches, without hemodynamic severity. A broad-spectrum probabilistic antibiotic therapy of ceftriaxone and metronidazole was started due to first blood cultures positive for anaerobic Gram-negative bacilli, while waiting for identification of the pathogen. Three days later, F. nucleatum was identified. According to the microbiological results, antibiotic therapy was adapted with amoxicillin and clavulanic acid, and no further complications were observed during clinical or complementary examinations. The final diagnosis was a F. nucleatum oropharyngeal infection complicated by bacteremia, without metastatic spread. Conclusion The etiologies of tonsillar infection are not limited to benign viruses or bacteria. These should not be overlooked in emergency medicine, especially when the clinical presentation is discrepant. A combination of early bacterial investigations as blood culture and close clinical monitoring is the only safe way to detect bacteremia, especially in immunocompetent patients.

A Comprehensive Method for the Evaluation of Hermetia illucens Egg Quality Parameters: Implications and Influence Factors

The significant momentum received by Hermetia illucens as a worldwide species is due to its biological traits and large applicability in scientific research, environmental entomoremediation, insect meal production, and for biodiesel yield. The aim of this research is to develop a method for the preparation and precise egg counting of the H. illucens egg clutch, as well as an accurate technique for evaluating egg biometric parameters. The precise proposed method for egg preparation and counting consists in dispersing the eggs clutch under a stereo microscope and counting the eggs on a photographic capture using the Clickmaster software. Five solution types were used to disperse the egg clutches: glycerin 50%, ethanol 70%, ethanol 80%, physiological serum 0.9% and purified water. The efficiency of the estimation method for eggs number evaluation was also tested by using the estimated egg weight as a conversion factor. The biometric parameters of single eggs (length and width) were determined using the free Toupview software. The precise method of egg preparation and counting allows for the registration of the eggs number manually identified by the operator. The appropriate dispersion solutions were glycerin 50% and ethanol 70%. The method has an error of 1.4 eggs for each 500 counted eggs, thus an accuracy of over 99.4%. The eggs number estimation method is not applicable without significant errors, the accuracy being less than 32%, due to egg heterogeneity in the clutch. Biometric parameters (length and width) are positively correlated with egg weight (r = 0.759) and with the number of eggs in the clutch (r = 0.645). In conclusion, the results clearly suggest the method of egg preparation and precise counting for an accurate evaluation of quality parameters of the H. illucens clutches, as well as the technique for evaluating egg biometric parameters.

Intestinal Protozoan Parasitosis in 4- and 11-year-old Children in Cunduana Community, Riobamba Canton, Chimborazo, June 2019 June

Introduction: Parasitosis is a public health problem that affects millions of people, school-age children are more vulnerable, affects their physical and intellectual development, are transmitted by fecal-oral route, and is associated with poor hygienic and sanitary conditions. In Ecuador they constitute the second reason for medical consultation. Objective: To contribute to the epidemiological knowledge of intestinal parasitosis by protozoa in our environment. Methodology: An experimental descriptive study, with a universe of 80 children, obtaining 55 samples, a direct examination was carried out in preparations with physiological serum and lugol, identifying the parasites by their microscopic morphology. Results: The prevalence of intestinal parasitosis by protozoa was 78.18%, while for the parasites Endolimax nana 30.9% Entamoeba histolytica, 34.5% and Giardia lamblia 14.5%. 40% of the participants were monoparasitized and 38.2% had polyparasitosis. Conclusion: High prevalence of parasitosis by intestinal protozoa was determined in the children of the Unit with 78.18%. The most prevalent ameboid protozoan was E. nana, followed by the pathogenic protozoan E. histolytica, while G. lamblia was flagellated protozoan identified. The lack of adequate sanitary infrastructure, habits of hygiene, environmental pollution, influence the prevalence of intestinal parasitosis. Keywords: protozoan infections, intestinal diseases, child. RESUMEN Introducción: Parasitosis es un problema de salud pública que afecta a millones de personas, los niños en edad escolar son más vulnerables, afecta su desarrollo físico e intelectual, se transmiten por vía fecal-oral, y está asociada a deficientes condiciones higiénicas y sanitarias. En Ecuador constituyen el segundo motivo de consulta médica. Objetivo: contribuir al conocimiento epidemiológico de las parasitosis intestinales por protozoos en nuestro medio. Metodología: estudio descriptivo experimental, con un universo de 80 niños y niñas, obteniendo 55 muestras, se realizó examen directo en preparaciones con suero fisiológico y lugol, identificando los parásitos por su morfología microscópica. Resultados: La prevalencia de parasitosis intestinal por protozoos fue de 78,18%, mientras que para los parásitos Endolimax nana 30,9%, Entamoeba histolytica 34,5% y Giardia lamblia 14,5%. El 40% de los participantes fue monoparasitado y el 38,2% presentó poliparasitosis. Conclusiones: Se determinó alta prevalencia de parasitosis por protozoos intestinales en los niños y niñas de la Unidad con un 78,18%. El protozoo ameboideo de mayor prevalencia fue E. nana, seguido del protozoo patógeno E. histolytica, mientras que G. lambia fue protozoo flagelado identificado. La falta de una adecuada infraestructura sanitaria, hábitos de higiene, contaminación ambiental, influyen en la prevalencia de la parasitosis intestinal. Palabras clave: infecciones por protozoos, parasitosis intestinales, niños.

P–613 Adjuvant letrozole in ovarian stimulation for in vitro fertilization does not reduce uterine peristalsis frequency prior to fresh embryo transfer

Study question Does adjuvant letrozole in ovarian stimulation (OS) for in vitro fertilization (IVF) decrease the uterine peristalsis frequency (UPF) prior to fresh embryo transfer (ET)? Summary answer Adjuvant letrozole in (OS) for IVF does not reduce the UPF significantly prior to fresh ET. What is known already Throughout the cycle UPF aids spermatozoa transport to the fallopian tube and may affect implantation. At fresh, ET UPF is negatively correlated with implantation- and clinical pregnancy rates and is believed to be modulated by estradiol and progesterone. High levels of estradiol, from multiple follicular development, in OS have been reported to increase UPF, whereas progesterone is considered to be utero-relaxant. The influence of androgens is unclear. Co-treatment with letrozole during gonadotropin OS limits the estradiol rise the supra-physiological estradiol and may therefore reduce UPF prior to fresh ET. Study design, size, duration: This single centre study was nested within a multicentre double blinded RCT investigating the impact of letrozole co-treatment during gonadotropin OS for IVF on late follicular and luteal estradiol, progesterone and testosterone levels. Between 2016 and 2017, 39 women expected normal responders were randomised to co-treatment with letrozole or placebo. Of these, 33 women completed this element of the study. The study was carried out according to the Helsinki Declaration and the ICH-Good-Clinical-Practice. Participants/materials, setting, methods Eligible women were randomised 1:1 to adjuvant treatment with letrozole 5 mg/day or placebo in an antagonist protocol using a fixed dose of recFSH 150 IU/day. Final maturation was triggered with rhCG 6,500 IU and luteal support with vaginal progesterone was administered from the day following oocyte aspiration. Less than one hour prior to fresh ET, six minute duration transvaginal ultrasound recordings of the uterus in sagittal section were performed and blood samples were drawn. Main results and the role of chance A total of 33 women completed the study (letrozole n = 17; placebo n = 16). Age, BMI, and ovarian reserve markers were similar between the groups. On day of ET, serum estradiol levels were significantly suppressed in the letrozole group to mean 867 ± 827 pmol/L compared to 3,110 ± 1,528 pmol/L in the placebo group (P < 0.0001). Mean UPF prior to fresh ET did not differ between the intervention and control group (3.3 ± 0.36 versus 3.5 ± 0.51 per minute respectively, P = 0.108). UPF was assessed and agreed by two observers who were blind to adjuvant treatment. Two patients were excluded due to poor quality of the ultra sound recording. Supra-physiological serum estradiol in the placebo group was negatively correlated with UPF (P = 0.014; R = –0.62), but the more physiological serum estradiol levels in the letrozole group showed no correlation with UPF (P = 0.567; R = 0.15). Serum progesterone levels were similar in both groups and did not show any significant correlation with UPF. Testosterone levels were significantly higher in the letrozole group (P = 0.005) and showed a non-significant trend negatively correlated with UPF in the placebo group (P-value=0.07, R= –0.48). Limitations, reasons for caution The limited sample size risks masking minor effects. Wider implications of the findings: The supra-physiological levels of estradiol were significantly supressed in the intervention group, but UPF prior to fresh ET was similar in both groups. UPF is not strongly correlated to luteal phase sex steroid levels. Any beneficial effect of adjuvant letrozole during OS is not through an impact of UPF. Trial registration number NCT02939898

Maintenance of muscle mass in adult male mice is independent of testosterone

Testosterone is considered a potent anabolic agent in skeletal muscle with a well-established role in adolescent growth and development in males. However, the role of testosterone in the regulation of skeletal muscle mass and function throughout the lifespan has yet to be fully established. While some studies suggest that testosterone is important for the maintenance of skeletal muscle mass, an understanding of the role this hormone plays in young, adult, and old males with normal and low serum testosterone levels is lacking. We investigated the role testosterone plays in the maintenance of muscle mass by examining the effect of orchiectomy-induced testosterone depletion in C57Bl6 male mice at ages ranging from early postnatal through old age (1.5-, 5-, 12-, and 24-month old mice). Following 28 days of testosterone depletion, we assessed mass and fiber cross-sectional-area (CSA) of the tibialis anterior, gastrocnemius, and quadriceps muscles. In addition, we measured global rates of protein synthesis and degradation using the SuNSET method, western blots, and enzyme activity assays. Twenty-eight days of testosterone depletion resulted in reduced muscle mass in the two youngest cohorts, but had no effect in the two oldest cohorts. Mean CSA decreased only in the youngest cohort and only in the tibialis anterior muscle. Testosterone depletion resulted in a general increase in proteasome activity at all ages. No change in protein synthesis was detected at the terminal time point. These data suggest that within physiological serum concentrations, testosterone may not be critical for the maintenance of muscle mass in mature male mice; however, in young mice testosterone is crucial for normal growth.

Evaluation of the flexural strength and surface roughness of dental material after immersion in coffee

This study was done in order to evaluate the changes in flexural strength and surface roughness in dental material restorations after immersion in coffee. Bars specimens (2mm x 2mm x 25 mm) of Z100 3M/ESPE were made according to ISO 4049 using aluminum molds. After curing, the specimens were then randomly divided into five groups and conditioned at 37°C in physiological serum. In all groups, except control, samples were immersed in coffee for ten minutes daily. In group A, the samples were immersed in coffee. For group B, after to be immersed in coffee, samples were immersed in distilled water for one minute. For group C, samples were immersed in mouthwash for one minute. For group D, samples were brushing for ten minutes, with load of 250g and 4250 cycles after all treatments the samples were stored in physiological serum again. In control group, samples were only stored in physiological serum. The five groups were then divided into 2 subgroups according storage time, six and nine weeks, respectively. Flexural strength was determined using three point bending test in a universal testing machine. All the tests were carried out at a room temperature and samples were maintained in physiological serum during the test. Surface roughness measurements were made using a surface roughness tester. All data were analyzed using one-way analysis of variance (ANOVA) followed by Dunnet tests. The ANOVA indicated no difference in the means of surface roughness and significant was observed for flexural strength between control group and one group (coffee).

TRPM4 Participates in Aldosterone-Salt-Induced Electrical Atrial Remodeling in Mice

Aldosterone plays a major role in atrial structural and electrical remodeling, in particular through Ca2+-transient perturbations and shortening of the action potential. The Ca2+-activated non-selective cation channel Transient Receptor Potential Melastatin 4 (TRPM4) participates in atrial action potential. The aim of our study was to elucidate the interactions between aldosterone and TRPM4 in atrial remodeling and arrhythmias susceptibility. Hyperaldosteronemia, combined with a high salt diet, was induced in mice by subcutaneously implanted osmotic pumps during 4 weeks, delivering aldosterone or physiological serum for control animals. The experiments were conducted in wild type animals (Trpm4+/+) as well as Trpm4 knock-out animals (Trpm4-/-). The atrial diameter measured by echocardiography was higher in Trpm4-/- compared to Trpm4+/+ animals, and hyperaldosteronemia-salt produced a dilatation in both groups. Action potentials duration and triggered arrhythmias were measured using intracellular microelectrodes on the isolated left atrium. Hyperaldosteronemia-salt prolong action potential in Trpm4-/- mice but had no effect on Trpm4+/+ mice. In the control group (no aldosterone-salt treatment), no triggered arrythmias were recorded in Trpm4+/+ mice, but a high level was detected in Trpm4-/- mice. Hyperaldosteronemia-salt enhanced the occurrence of arrhythmias (early as well as delayed-afterdepolarization) in Trpm4+/+ mice but decreased it in Trpm4-/- animals. Atrial connexin43 immunolabelling indicated their disorganization at the intercalated disks and a redistribution at the lateral side induced by hyperaldosteronemia-salt but also by Trpm4 disruption. In addition, hyperaldosteronemia-salt produced pronounced atrial endothelial thickening in both groups. Altogether, our results indicated that hyperaldosteronemia-salt and TRPM4 participate in atrial electrical and structural remodeling. It appears that TRPM4 is involved in aldosterone-induced atrial action potential shortening. In addition, TRPM4 may promote aldosterone-induced atrial arrhythmias, however, the underlying mechanisms remain to be explored.

Histopathological evaluation of Senecio rhizomatus Rusby in 7,12-dimethylbenz(α) anthracene-induced breast cancer in female rats

Background and Aim: Senecio rhizomatus Rusby (SrR) is a medicinal plant of the Asteraceae family and traditionally consumed as infusion in the Andean region from Peru for inflammatory disorders. This study aimed to determine the histopathological changes afforded by SrR in 7, 12-dimethylbenz[α]anthracene (DMBA)-induced breast cancer (BC) in rats. Materials and Methods: An ethanolic extract of SrR aerial parts was prepared by maceration with 96% ethanol, and the chemical components were identified by gas chromatography coupled to mass spectrometry; the antioxidant activity was determined by 1,1-diphenyl-2-picril-hidrazil (DPPH) assay; and the acute toxicity was assessed according to the OCED 423 guidelines. In a pharmacological study, 30 female Holztman rats were distributed randomly into five groups, as follows. Group I: Negative control (physiological serum, 2 mL/kg); Group II. DMBA (80 mg/Kg body weight); and Groups III, IV, and V: DMBA + ethanol extract of SrR at doses of 10, 100, and 200 mg/kg, respectively. Results: The antioxidant activity of the SrR extract against DPPH was 92.50% at 200 μg/mL. The oral administration of SrR at doses of 50, 300, 2000, and 5000 mg/kg did not show any clinical evidence of toxicity or occurrence of death. The groups that received SrR presented a lower frequency of tumors and a cumulative tumor volume compared with the DMBA group (p<0.05); the DMBA group exhibited a higher incidence of necrosis and moderate mitosis, up to 66.67% and 100.00%, respectively. Finally, infiltrating carcinoma with extensive tumor necrosis was evidenced. Conclusion: In experimental conditions, the ethanolic extract of SrR had a protective effect in DMBA-induced BC in female rats. Furthermore, the antioxidant activity of its main phytochemicals could be responsible for the effect observed, and SrR seems to be a safe extract in the preclinical phase.

Role of Testosterone Levels on the Combinatorial Effect of Boswellia serrata Extract and Enzalutamide on Androgen Dependent LNCaP Cells and in Patient Derived Cells

Co-therapy with herbal extracts along with current clinical drugs is being increasingly recognized as a useful complementary treatment for cancer. The anti-cancer property of the phyto-derivative acetyl-11 keto β boswellic acid (AKBA) has been studied in many cancers, including prostate cancer. However, the whole extract of the gum resin Boswellia serrata (BS) and anti-androgen enzalutamide has not been explored in prostate cancer to date. We hypothesized that the BS extract containing 30% (AKBA) with enzalutamide acted synergistically in the early phase of cancer, especially in LNCaP cells, by inhibiting androgen receptor (AR) and by reducing cell proliferation, and further, that the extract would be superior to the action of the active ingredient AKBA when used alone or in combination with enzalutamide. To test our hypothesis, we treated LNCaP cells with BS extract or AKBA and enzalutamide both individually and in combination to analyze cell viability under different levels of dihydrotestosterone (DHT). The inhibition of androgen receptor (AR) followed by the expression of prostate-specific antigen (PSA) and the efflux mechanism of the cells were analyzed to determine the effect of the combination on the cellular mechanism. Cells derived from prostate cancer patients were also tested with the combination. Only 6 µM enzalutamide along with BS in the range of 4.1 µg/ml to 16.4 µg/ml gave the best synergistic results with nearly 50% cell killing even though standard enzalutamide doses were as high as 48 µM. Cell killing was most effective at intermediate DHT concentrations of approximately 1 nM, which corresponds to normal physiological serum levels of DHT. The Pgp expression level and the androgen receptor expression levels were reduced under the combination treatment; the former helping to minimize drug efflux and the latter by reducing the sensitivity to hormonal changes. Furthermore, the combination reduced the PSA level secreted by the cells. In contrast, AKBA could not achieve the needed synergism for adequate cell killing at equivalent concentrations. The combination of enzalutamide and BS extract containing 30% AKBA because of their synergistic interaction is an attractive therapeutic option for treating early stage (hormone-dependent) prostate cancer and is superior to the use of AKBA alone.