A fast and inexpensive genotyping system for the simultaneous analysis of human and Aedes albopictus short tandem repeats.

BACKGROUND: Determination of the interactions between hematophagous mosquitoes and their human hosts is of great importance for better understanding the transmission dynamics of mosquito-borne arboviruses and developing effective strategies to mitigate risk. Genetic analysis of human and mosquito DNA can play a key role in this, but commercial kits for human short tandem repeat (STR) genotyping are expensive and do not allow for the simultaneous STR analysis of host and vector DNA. Here, we present an inexpensive and straightforward STR-loci multiplex system capable of simultaneously amplifying Aedes albopictus and human STRs from blood-fed mosquitoes. Additionally, we examine the effect of storage methods and post-feeding time on the integrity of host DNA. METHODS: Thirty-five STRs (16 human and 19 Ae. albopictus STRs) subdivided in three multiplexes were tested for amplification and scoring reliability. Under laboratory conditions we compared the efficacy of two preservation methods (absolute ethanol vs lysis buffer) on the integrity of host DNA in Ae. albopictus blood meals. We also evaluated the effect of post-feeding time by sacrificing blood-fed mosquitoes at different time intervals after feeding, and we assessed our ability to detect multiple feedings. To determine if the system can be employed successfully under field conditions, we carried out a preliminary study using field-collected Ae. albopictus. RESULTS: All 35 STRs amplified consistently in the laboratory. Lysis buffer performed better than absolute ethanol in terms of allele peak height and clarity of electropherograms. Complete human DNA profiles could be obtained up to 48 h following the blood meal. Analysis of multiple feedings confirmed that peak heights can be used as a proxy to determine post-feeding time and thus derive the number of different people bitten by a mosquito. In the field trial, amplification was successful for 32 STRs. We found human DNA signal in 38 of the 61 field-collected mosquitoes (62%), of which 34 (89%) had ingested a single blood meal, while four (11%) contained double meals. CONCLUSIONS: Our new genotyping system allows fast and reliable screening of both host and vector species, and can be further adapted to other mosquito species living in close contact with humans.

Drivers of longevity of wild-caught Aedes albopictus populations.

BACKGROUND: Age structure and longevity constitute fundamental determinants of mosquito populations' capacity to transmit pathogens. However, investigations on mosquito-borne diseases primarily focus on aspects such as abundance or dispersal rather than survival and demography. Here, we examine the post-capture longevity of wild-caught populations of the Asian tiger mosquito Aedes albopictus to investigate the influence of environmental factors and individual frailty on longevity. METHODS: We captured females of Ae. albopictus from June to November 2021 in a vegetated and an urban area by two methods of capture (BG traps and Human Landing catch). They were kept in semi-controlled conditions in the field, and survival was monitored daily across the 859 individuals captured. We studied the differences in longevity per capture method and location and the influence on longevity of seasonal, climatic and individual factors. RESULTS: Photoperiod, GDD, minimum and maximum temperature and relative humidity showed an effect on the risk of death of females in the field. Females captured in urban area with Human Landing catch methods had greater longevity than females captured in non-urban areas with BG traps. Individual variance, reflecting individual frailties, had an important effect on the risk of death: the greater the frailty, the shorter the post-capture longevity. Overall, longevity is affected not only by climate and seasonal drivers like temperature and photoperiod but also by the individual frailty of mosquitoes. CONCLUSION: This work unravels environmental drivers of key demographic parameters such as longevity, as modulated by individual frailty, in disease vectors with strong seasonal dynamics. Further demographic understanding of disease vectors in the wild is needed to adopt new surveillance and control strategies and improve our understanding of disease risk and spread.

Grazing in the dark: A behavioural adjustment in a population of the black sea urchin Arbacia lixula.

In Mediterranean rocky shores, the black sea urchin Arbacia lixula is often associated with communities dominated by encrusting corallines, devoid of fleshy algae. While it is commonly known as a diurnal herbivore, this species also migrates at night from hidden to more exposed habitats. Here, we provide the first experimental evidence of an adjustment to a predominant nocturnal behaviour in a population of A. lixula. Sea urchin densities changed from nearly zero during daytime to more than 16 urchins m-2 at night in treatment plots where the sea urchins were removed. We suggest that the observed behaviour was triggered by our experimental manipulations and was a response to the presence of dead conspecifics and small predatory fishes attracted by the urchin culling. Further research is needed to assess whether our findings can be generalised to the behaviour of A. lixula in areas where sea urchins are under strong pressure from diurnal predators. In these cases, it is important to perform sea urchin density counts at night to avoid misleading assessments about the herbivore pressure in a littoral area.

Multiple invasions, Wolbachia and human-aided transport drive the genetic variability of Aedes albopictus in the Iberian Peninsula.

The Asian tiger mosquito, Aedes albopictus, is one of the most invasive species in the world. Native to the tropical forests of Southeast Asia, over the past 30 years it has rapidly spread throughout tropical and temperate regions of the world. Its dramatic expansion has resulted in public health concerns as a consequence of its vector competence for at least 16 viruses. Previous studies showed that Ae. albopictus spread has been facilitated by human-mediated transportation, but much remains unknown about how this has affected its genetic attributes. Here we examined the factors that contributed to shaping the current genetic constitution of Ae. albopictus in the Iberian Peninsula, where the species was first found in 2004, by combining population genetics and Bayesian modelling. We found that both mitochondrial and nuclear DNA markers showed a lack of genetic structure and the presence of worldwide dominant haplotypes, suggesting regular introductions from abroad. Mitochondrial DNA showed little genetic diversity compared to nuclear DNA, likely explained by infection with maternally transmitted bacteria of the genus Wolbachia. Multilevel models revealed that greater mosquito fluxes (estimated from commuting patterns and tiger mosquito population distribution) and spatial proximity between sampling sites were associated with lower nuclear genetic distance, suggesting that rapid short- and medium-distance dispersal is facilitated by humans through vehicular traffic. This study highlights the significant role of human transportation in shaping the genetic attributes of Ae. albopictus and promoting regional gene flow, and underscores the need for a territorially integrated surveillance across scales of this disease-carrying mosquito.

MRI and MR voiding cystourethrography in the evaluation of male primary bladder neck obstruction: preliminary experience.

PURPOSE: Evaluation of male with primary bladder neck obstruction (PBNO) using MRI and MR voiding cystourethrography (MR-VCU) to study both anatomical aspects of bladder neck and urethral lumen. METHODS: In this retrospective study 21 male patients (mean age 33 ± 14) with urodynamic diagnosis of PBNO and 5 healthy volunteers ((mean age 28 ± 2) as control group were enrolled. Both patients and control group underwent 1.5 T MRI. Sagittal and oblique coronal Turbo-Spin-Echo T2-weighted scans were performed. Only patients underwent MR voiding cystourethrography (MR-VCU) performed with T1-weighted spoiled 3D gradient-echo sagittal acquisitions. Bladder lumen was filled with contrast-material-enhanced urine. Blinded test by two radiologists was performed to evaluate causes of bladder outlet obstruction evaluating MR-VCU. Anatomical MRI features of both control group and patients were compared in consensus by senior radiologist and urologist using the analysis of variance (ANOVA) test. RESULTS: MRI allowed evaluation of the bladder neck muscular structures. We found 4 groups of PBNO patients: 52% hypertrophy of posterior lip of bladder sphincter; 20% asymmetry of lateral portion of bladder sphincter; 14% bladder neck cyst; 14% showed normal aspect of bladder neck. Comparison between the control group and first and second PBNO groups was considered statistically significant (p < 0.05) with diagnostic accuracy of 87%. Only 13 patients (61%) were able to perform MR-VCU and radiologists always made the diagnosis of PBNO. CONCLUSION: MRI together with MR-VCU provides useful anatomical and functional information in the study of bladder neck and urethral lumen. These preliminary results suggest that MRI could substitute for standard cystourethrogram in patients with PBNO.

Ecological traits, genetic diversity and regional distribution of the macroalga Treptacantha elegans along the Catalan coast (NW Mediterranean Sea).

The widespread decline of canopy-forming macroalgal assemblages has been documented in many regions during the last decades. This pattern is often followed by the replacement of structurally complex algal canopies by more simplified habitats (e.g., turfs or sea urchin barren grounds). Against all odds, the fucoid Treptacantha elegans, a large Mediterranean brown macroalga, broadened its depth range to deeper and exposed environments and displayed an unexpected range expansion along the northern coast of Catalonia over the last two decades. Here, we reconstruct the spread of T. elegans in time and space and unravel ecological and demographic traits such as population dynamics and genetic patterns to provide a comprehensive and integrated view of the current status and geographical expansion for this species. Fast-growing dynamics, early fertile maturity, and high turnover rate are the main competitive advantages that allow the exposed populations of T. elegans to colonize available substrata and maintain dense and patchy populations. We also provided evidence that the deeper and exposed populations of T. elegans constitute a single group across the Catalan coast, with little genetic differentiation among populations. This seems to support the hypothesis of a unique source of spread in the last decades from the Medes Islands No-Take Zone towards both southern and northern waters.

Once-daily 5 mg tadalafil oral treatment for patients with chronic prostatitis/chronic pelvic pain syndrome.

BACKGROUND: Chronic prostatitis/chronic pelvic pain syndrome (IIIB CP/CPPS) is a condition of unclear aetiology. Many approaches have been used without satisfactory results. The aim of this study is to evaluate the efficacy of once-daily 5 mg tadalafil in pain control and improving quality of life in patients affected by CP/CPPS. METHODS: Twenty patients affected by chronic prostatitis according EAU (European Association of Urology) guidelines were evaluated for once-daily 5 mg tadalafil; 14 patients were eligible for the study. The validated Italian version of the NIH Chronic Prostatitis Symptom Index (NIH-CPSI) and the International Prostatic Symptom Score (IPSS) questionnaires were submitted to all the patients. Every patient underwent uroflowmetry and ultrasound prostatic volume at the beginning and at the end of the study. RESULTS: All 14 patients eligible for the study reported an improvement of symptoms during therapy: statistically significant differences were reported in terms of NIH-CPSI (p < 0.000002) and IPSS (p < 0.0001) during follow-up evaluations. No statistically significant improvement of uroflowmetry parameters was reported during the treatment. CONCLUSIONS: In our study the daily use of 5 mg tadalafil improves symptoms and quality of life in patients affected by CP/CPPS after 4 weeks of therapy. A larger population of patients is needed to confirm the efficacy of this therapy in CP/CPPS.

The optimal sampling design for littoral habitats modelling: A case study from the north-western Mediterranean.

Species distribution models (SDMs) have been used to predict potential distributions of habitats and to model the effects of environmental changes. Despite their usefulness, currently there is no standardized sampling strategy that provides suitable and sufficiently representative predictive models for littoral marine benthic habitats. Here we aim to establish the best performing and most cost-effective sample design to predict the distribution of littoral habitats in unexplored areas. We also study how environmental variability, sample size, and habitat prevalence may influence the accuracy and performance of spatial predictions. For first time, a large database of littoral habitats (16,098 points over 562,895 km of coastline) is used to build up, evaluate, and validate logistic predictive models according to a variety of sampling strategies. A regularly interspaced strategy with a sample of 20% of the coastline provided the best compromise between usefulness (in terms of sampling cost and effort) and accuracy. However, model performance was strongly depen upon habitat characteristics. The proposed sampling strategy may help to predict the presence or absence of target species or habitats thus improving extensive cartographies, detect high biodiversity areas, and, lastly, develop (the best) environmental management plans, especially in littoral environments.

Low prostate concentration of lycopene is associated with development of prostate cancer in patients with high-grade prostatic intraepithelial neoplasia.

Prostate cancer (PC) is a frequent male malignancy and represents the second most diagnosed cancer in men. Since pre-cancerous lesions, i.e., the high-grade prostatic intraepithelial neoplasia (HGPIN), can be detected years before progression to PC, early diagnosis and chemoprevention are targeted strategies to reduce PC rates. Animal studies have shown that lycopene, a carotenoid contained in tomatoes, is a promising candidate for the chemoprevention of PC. However, its efficacy in humans remains controversial. The present study aimed to investigate the relevance of plasma and prostate concentration of lycopene after a lycopene-enriched diet in patients diagnosed with HGPIN. Thirty-two patients diagnosed with HGPIN were administered a lycopene-enriched diet (20-25 mg/day of lycopene; through 30 g/day of triple concentrated tomato paste) for 6 months. A 6-month follow-up prostate biopsy assessed progression to PC. Patients were classified into three groups according to the histopathological features of the 6-month follow-up biopsy results: prostatitis; HGPIN and PC. PSA and plasma lycopene levels were measured before and after the dietary lycopene supplementation. Prostatic lycopene concentration was only assessed after the supplementation diet. Only prostatic lycopene concentration showed significant differences between the three groups (p = 0.03). Prostatic lycopene concentration below a 1 ng/mg threshold was associated with PC at 6-month follow-up biopsy (p = 0.003). We observed no overall benefits from a 6-month lycopene supplementation, as the rate of HGPIN progression to PC in our population (9/32, 28%) was similar to rates reported in the literature. Baseline PSA levels also showed no significant changes after a lycopene-enriched diet. Our findings point to prostatic lycopene concentration as a promising biomarker of PC. Further prospective longitudinal studies are needed to assess the prognostic role of prostatic lycopene in PC.

Transurethral resection of prostate and the role of pharmacological treatment with dutasteride in decreasing surgical blood loss.

PURPOSE: Transurethral resection of prostate (TURP) still represents the gold standard in the surgical treatment of symptomatic benign prostatic hyperplasia (BPH). The most frequent complication is represented by intra- and perioperative bleeding. Preoperative use of 5-alpha-reductase inhibitors (finasteride or dutasteride) to reduce surgical bleeding is still a topic of debate in literature. Previous studies provided favorable data on blood loss reduction by preoperative administration of finasteride or dutasteride. The aim of this study was to evaluate whether pretreatment with dutasteride for six weeks before surgery can reduce surgical blood loss. METHODS: A total of 142 patients with BPH-who were to undergo TURP-were enrolled and randomized into two groups. The dutasteride group comprising of 71 patients, was treated with dutasteride (0.5 mg/day) for 6 weeks before surgery and the control group, comprising of other 71 patients, did not receive dutasteride. Blood loss was evaluated in terms of a reduction in the serum hemoglobin level (ΔHb and ΔHCT), and was estimated by measuring the Hb and hematocrit levels before and 24 hours after surgery. RESULTS: None of the patients treated with dutasteride reported any side effects. A significantly lower mean blood loss was observed in the dutasteride group compared to the control group (ΔHb=-1.29 ± 0.81 v -1.83 ± 1.25, respectively, p<0.0027; ΔHCT=-5.67 ± 2.58 v -6.50 ± 2.40, respectively, p<0.0491). CONCLUSIONS: Our results showed that pretreatment with dutasteride for 6 weeks before TURP reduces the surgical bleeding considerably. This treatment schedule can be used routinely to decrease TURP surgical bleeding.

The structure of Mediterranean rocky reef ecosystems across environmental and human gradients, and conservation implications.

Historical exploitation of the Mediterranean Sea and the absence of rigorous baselines makes it difficult to evaluate the current health of the marine ecosystems and the efficacy of conservation actions at the ecosystem level. Here we establish the first current baseline and gradient of ecosystem structure of nearshore rocky reefs at the Mediterranean scale. We conducted underwater surveys in 14 marine protected areas and 18 open access sites across the Mediterranean, and across a 31-fold range of fish biomass (from 3.8 to 118 g m(-2)). Our data showed remarkable variation in the structure of rocky reef ecosystems. Multivariate analysis showed three alternative community states: (1) large fish biomass and reefs dominated by non-canopy algae, (2) lower fish biomass but abundant native algal canopies and suspension feeders, and (3) low fish biomass and extensive barrens, with areas covered by turf algae. Our results suggest that the healthiest shallow rocky reef ecosystems in the Mediterranean have both large fish and algal biomass. Protection level and primary production were the only variables significantly correlated to community biomass structure. Fish biomass was significantly larger in well-enforced no-take marine reserves, but there were no significant differences between multi-use marine protected areas (which allow some fishing) and open access areas at the regional scale. The gradients reported here represent a trajectory of degradation that can be used to assess the health of any similar habitat in the Mediterranean, and to evaluate the efficacy of marine protected areas.

Reduction of prostate-specific antigen after tamsulosin treatment in patients with elevated prostate-specific antigen and lower urinary tract symptoms associated with low incidence of prostate cancer at biopsy.

OBJECTIVES: To evaluate the effect of tamsulosin on reducing the serum levels of prostate-specific antigen (PSA) in patients with lower urinary tract symptoms and an elevated PSA level. METHODS: From June 2004 to July 2006, patients with lower urinary tract symptoms, a PSA level of >or=4 ng/mL, and a maximal flow rate of <15 mL/s received tamsulosin 0.4 mg daily for 2 months. They were then scheduled for 12-core prostate biopsy. PSA determination and a uroflow study were performed before biopsy. RESULTS: A total of 80 patients completed the present study. The mean patient age was 66.3 years, and the mean PSA level was 7.8 +/- 8.4 ng/mL at baseline and 7.1 +/- 9.1 ng/mL after treatment (P < .001). A total of 29 patients (36.25%) were diagnosed with prostate cancer from the biopsy findings. A significant increment in the PSA level was observed in patients with prostate cancer (6.7 versus 7.9 ng/mL; P = .002). A significant decrease in the PSA level was observed in patients with negative biopsy findings (6.9 versus 5.1 ng/mL, P = .000). Of the 38 patients with a decrease in the PSA level, 1 (2.6%) was diagnosed with prostate cancer and 37 (97.4%) with an benign prostatic hyperplasia/prostatitis. Of the 42 patients with no change in the PSA level, 28 (66.7%) had prostate cancer and 14 (33.3%) had negative findings. A change in PSA level after treatment gave a sensitivity of 96.6%, specificity of 72.5%, and diagnostic accuracy of 81% for prostate cancer. CONCLUSIONS: Treatment with tamsulosin seemed to reduce the PSA levels and identified patients at high risk of prostate cancer.

Bladder weight and detrusor thickness as parameters of progression of benign prostatic hyperplasia.

PURPOSE OF REVIEW: This manuscript reviews the information that became available over the last 12 months on the measurement of bladder/detrusor wall thickness and ultrasound-estimated bladder weight in patients with benign prostatic enlargement. The subject is of particular interest because novel technological developments may help to standardize and automate the measurement technique. RECENT FINDINGS: Preliminary data on the automatic measurement of bladder wall thickness were reported, suggesting a good repeatability and agreement with conventional ultrasound imaging, although variance is higher than with conventional ultrasound, and thicknesses above 4 mm were not measured correctly. Some controversies emerged in the most recent peer-review literature on the possibility to diagnose bladder outlet obstruction and detrusor overactivity by measuring bladder wall thickness, and this certainly pleaded for further collaborative research and standardization. Preliminary results from new technological developments suggest a very accurate measure of bladder volume and good repeatability of bladder wall and bladder weight measurements. CONCLUSION: After a decade of research, automated measurement of bladder is here to stay. Standardization of bladder wall and weight measurement will allow better understanding of the physiopathology of detrusor hypertrophy in lower urinary tract dysfunction and possibly provide useful prognosticator indexes for our daily practice.

Through the explanatory process in natural history and ecology.

In this paper I will deal with the explanatory process used in natural history and ecology. I argue that the development of knowledge in natural history and descriptive ecology is the result of a bottom-up process, which is mainly empirical and progresses continuously from entity perception to theory construction. I consider the role of observation in the development of abstract images of entities, patterns, and processes through concepts and theories from a "simple" cognitive point of view and without regard for computational aspects. I analyze whether natural history provides "real" scientific explanation or just mere observation of facts. I later discuss the role of principles and laws in the explanation of observed regularities and accidents and the importance of prediction. I use the study of the larvae of sponges to describe the process because they represent a good example of past and current scientific method. My main argument is pragmatic being that the only relevant matters of the explanatory process are the perspective from which we observe the facts, the categorization methods we are using, and an acknowledgement of their scientific rigor. We need to advance in our epistemology in order to capture all the different meanings that the word "science" has acquired rather than sticking to one dominated by currently accepted methodologies.

Dispersal strategies in sponge larvae: integrating the life history of larvae and the hydrologic component.

While known to be uniformly non-feeding, short-lived, and potentially short dispersing, sponge larvae display different behaviours (swimming ability and taxis). Our aim was to show whether sponge larvae with different behaviours exhibit different dispersal strategies under variable intensity of water movements. We first assessed the distribution of larvae of six taxa: Dictyoceratida spp., Dysidea avara, Crambe crambe, Phorbas tenacior, Scopalina lophyropoda, and Cliona viridis, collected through plankton sampling, and the abundance of the corresponding adult sponges across three hard bottom communities and a sandy bottom from a north-west Mediterranean rocky shore. We then tested adult-larvae couplings (abundance of larvae vs abundance of adults) under increasing levels of water movements (surge) to assess the importance of this environmental factor in driving differences in dispersal strategies. Adults of Dictyoceratida spp., D. avara, and P. tenacior were most abundant in semi-dark caves (SDC), C. crambe and C. viridis in communities of sciaphilic algae (SA), whereas the distribution of S. lophyropoda was extremely patchy, being present almost only in the SA community of one of the five stations studied. Larvae of Dictyoceratida spp. and P. tenacior were more abundant in the SDC, whereas D. avara and C. crambe were homogeneously distributed across the communities. The larvae of C. viridis were more abundant in the SA communities and the S. lophyropoda larvae were mostly present in one station and one community (SA). Increased water movement did not modify the adult-larvae coupling for Dictyoceratida spp., D. avara, and C. crambe, whereas it broke up the positive association for P. tenacior and to some extent S. lophyropoda. For C. viridis, possible variability in adult-larvae coupling was not tested because the larvae were collected on only one day under calm sea conditions. We confirm that efficient-swimming larvae with some cue response can actively counteract hydrodynamic forces and highlight the importance of both larval behaviour and environmental conditions in determining small-scale patterns of dispersal.

Local anesthesia reduces pain associated with transrectal prostatic biopsy. A prospective randomized study.

INTRODUCTION: To test the hypothesis that periprostatic block could completely relief prostatic biopsy-associated pain. MATERIALS AND METHODS: Patients scheduled for transrectal ultrasound guided prostate biopsy were randomized (1:1:1 ratio) to no analgesia (group A), endorectal enema of 1% lidocaine gel (group B) or transrectal periprostatic block (group C). All patients underwent 10 core TRUS-guided biopsy. After the procedure, a ten visual analogue pain score (VAS) from 0 = no discomfort to 10 = severe pain was administered to the biopsied patients and a global estimation of pain associated with the procedure was obtained. The study design included interim analysis of pain score after the first 60 patients were enrolled. Kruskal-Wallis test for unpaired data was used for statistical analysis. Data are presented as mean, median (range). RESULTS: Sixty patients were enrolled between May 2003 and December 2003 and all patients were evaluable. Mean and median age was 68.5 and 69 (range 53-82) years, respectively. Mean and median PSA was 86.8 and 9 ng/ml (range 0.58-4.111), respectively. No major side effects were observed. Patients in group A scored at VAS a median 4, mean 5.5 +/- 2.3 (range 3-10). Patients in group B scored a median 4, mean 5.5 +/- 2.7 (range 3-10) (p = 0.237). Patients receiving periprostatic injections of carbocaine (group C) scored a median 0, mean 0.5 +/- 0.8 (range 0-2). The level of pain reported by this group of patients was significantly different from those reported by patients who performed prostatic biopsy without anesthesia or with intrarectal anesthetic jelly (p = 0.00001). In the periprostatic block group 65% of patients referred no pain after the procedure (VAS = 0) while all patients in the other groups experience some degree of pain. CONCLUSION: The use of bilateral periprostatic block is a very effective and useful technique, well tolerated by the patient, which almost completely abolishes the pain and discomfort associated with the prostatic biopsy procedure.