Gene expression and cellular changes in injured myocardium of Ciona intestinalis.

Ciona intestinalis is an invertebrate animal model system that is well characterized and has many advantages for the study of cardiovascular biology. The regulatory mechanisms of cardiac myocyte proliferation in Ciona are intriguing since regeneration of functional tissue has been demonstrated in other organs of Ciona in response to injury. To identify genes that are differentially expressed in response to Ciona cardiac injury, microarray analysis was conducted on RNA from adult Ciona hearts with normal or damaged myocardium. After a 24- or 48-h recovery period, total RNA was isolated from damaged and control hearts. Initial results indicate significant changes in gene expression in hearts damaged by ligation in comparison to control hearts. Ligation injury shows differential expression of 223 genes as compared to control with limited false discovery (5.8%). Among these 223 genes, 117 have known human orthologs of which 68 were upregulated and 49 were downregulated. Notably, Fgf9/16/20, insulin-like growth factor binding protein and Ras-related protein Rab11b were significantly upregulated in injured hearts, whereas expression of a junctophilin ortholog was decreased. Histological analyses of injured myocardium were conducted in parallel to the microarray study which revealed thickened myocardium in injured hearts. Taken together, these studies will connect differences in gene expression to cellular changes in the myocardium of Ciona, which will help to promote further investigations into the regulatory mechanisms of cardiac myocyte proliferation across chordates.

Long-term urological outcome of cloaca patients with multidisciplinary management.

PURPOSE: Urological management of Cloacal Malformation (CM) focuses on preserving renal function and continence. Study aim was to analyze urinary and intestinal outcomes in CM patients, considering the length of common channel (CC) and presence of occult spinal dysraphism (OSD). METHODS: Retrospective review of CM treated at our institution by a multidisciplinary team from 1999 to 2020. Patients with follow-up < 2.5 years were excluded. Length of CC, renal function, urinary and bowel outcomes, presence of associated anomalies (especially OSD) were evaluated. RESULTS: Twenty patients were included, median age at follow-up: 8 years (4-15). A long CC > 3 cm was described in 11 (55%). Chronic kidney disease was found in 3 patients. Urinary continence was achieved in 8/20 patients, dryness (with intermittent catheterization) in 9/20. Fecal continence was obtained in 3/20, cleanliness in 14 (under bowel regimen). OSD was present in 10 patients (higher prevalence in long-CC, 73%). Among OSD, 1 patient reached fecal continence, 7 were clean; 2 achieved urinary continence, while 6 were dry. CONCLUSIONS: Length of CC and OSD may affect urinary and fecal continence. An early counseling can improve outcome at long-term follow-up. Multidisciplinary management with patient centralization in high grade institutions is recommended to achieve better results.

Hydrocarbons removal from real marine sediments: Analysis of degradation pathways and microbial community development during bioslurry treatment.

In this study, real marine sediments polluted by petroleum compounds were treated by means of a bioslurry pilot scale reactor. The treatment performance was evaluated by measuring the removal of total petroleum hydrocarbon (TPH), coupled to further analyses required to understand the mechanisms involved in the biodegradation process. The maximum TPH-removal efficiency reached 86 % at the end of experiments. Moreover, high throughput 16S RNA gene sequencing was used to describe the microbiome composition in sediment prior to, and after, bioslurry treatment, in order to identify the taxa mostly entailed in the TPH removal process. The raw sediment was mostly colonized by members of Sulfurimonas genus; after bioslurry treatment, it was noticed a shift in the microbial community composition, with Proteobacteria phylum dominating the remediation environment (high increase in terms of growth for Hydrogenophaga and Sphingorhabdus genera) along with the Phaeodactylibacter genus (Bacteroidetes). Furthermore, the assessment of gaseous emissions from the system allowed to quantify the volatile hydrocarbon component and, consequently, to obtain a more accurate evaluation of TPH-removal pathway by the bioslurry system. Finally, phytotoxicity tests on sediment samples highlighted an increase of the treated sample quality status compared to the untreated one.

Serial scanning with 99mTc-3, 3-diphosphono-1, 2-propanodicarboxylic acid (99mTc-DPD) for early detection of cardiac amyloid deposition and prediction of clinical worsening in subjects carrying a transthyretin gene mutation.

BACKGROUND: To determine the capability of 99mTc-DPD scintigraphy to detect early cardiac involvement and predict clinical worsening in transthyretin (TTR) gene mutation patients. METHODS: Eleven mutated subjects with normal interventricular septum (IVS) thickness, NT-proBNP level and no cardiac symptoms underwent three seriate 99mTc-DPD scans (visually and semiquantitatively analyzed), and was followed-up for 5-8-years. RESULTS: Six patients showed no myocardial accumulation in all scans. Increased IVS thickness occurring in one patient 4 years after the last scan was the only abnormal finding in these patients; no cardiac symptoms developed during the follow-up. In three patients, cardiac radiotracer uptake was found at enrollment; other laboratory/instrumental abnormal findings occurred later and cardiac symptoms developed during the follow-up period. Two patients had a negative 99mTc-DPD scan at enrollment and showed cardiac uptake in the following scans. Increased mean left-ventricular (LV) wall thickness was found 3 years after positive scintigraphy; NT-proBNP increased later in one patient. These patients developed cardiac symptoms during the follow-up period. CONCLUSIONS: 99mTc-DPD scan detects cardiac involvement in subjects with TTR gene mutation earlier than ECG, echocardiography and biochemical markers, occurring some years before the fulfillment of current diagnostic criteria for cardiac amyloidosis. A positive 99mTc-DPD scan predicts cardiac symptoms onset.

SARS-CoV-2 concentrations and virus-laden aerosol size distributions in outdoor air in north and south of Italy.

The COVID-19 disease spread at different rates in the different countries and in different regions of the same country, as happened in Italy. Transmission by contact or at close range due to large respiratory droplets is widely accepted, however, the role of airborne transmission due to small respiratory droplets emitted by infected individuals (also asymptomatic) is controversial. It was suggested that outdoor airborne transmission could play a role in determining the differences observed in the spread rate. Concentrations of virus-laden aerosol are still poorly known and contrasting results are reported, especially for outdoor environments. Here we investigated outdoor concentrations and size distributions of virus-laden aerosol simultaneously collected during the pandemic, in May 2020, in northern (Veneto) and southern (Apulia) regions of Italy. The two regions exhibited significantly different prevalence of COVID-19. Genetic material of SARS-CoV-2 (RNA) was determined, using both real time RT-PCR and ddPCR, in air samples collected using PM10 samplers and cascade impactors able to separate 12 size ranges from nanoparticles (diameter D < 0.056 µm) up to coarse particles (D > 18 µm). Air samples tested negative for the presence of SARS-CoV-2 at both sites, viral particles concentrations were <0.8 copies m-3 in PM10 and <0.4 copies m-3 in each size range investigated. Outdoor air in residential and urban areas was generally not infectious and safe for the public in both northern and southern Italy, with the possible exclusion of very crowded sites. Therefore, it is likely that outdoor airborne transmission does not explain the difference in the spread of COVID-19 observed in the two Italian regions.

Optimization of management choices of clariflocculation process by means of qualitative multi-criteria analysis.

Every year ship traffic produces tons of liquid waste mainly consisting of bilge water and of washing water of tankers' tanks. The latter are called slop waters and are characterized by high salinity and by the presence of recalcitrant pollutants mainly of hydrocarbon origin: these characteristics promote the use of chemical-physical rather than biological treatment. In particular, in the present study the slop waters were subjected to a clariflocculation treatment by means of batch tests. This treatment involves the dosage of specific chemical reagents (coagulants and flocculants) added to water at different stages of the process. In order to establish the optimal reagents' type and dose, also considering the operating costs, the proposed study presents a frequency analysis belonging to the family of multi-criteria exploration. The application of this methodology to examine the validity of the different process alternatives has allowed the inclusion of, in a single assessment, both economic and extra-economic (measurable only in qualitative terms) procedures. Thanks to this qualitative and quantitative method, it was therefore possible to order the different treatment alternatives analyzed, identifying the one that allows optimizing the wastewater management, for a conscious choice of the most suitable solution to the problem.

Does the feeding strategy enhance the aerobic granular sludge stability treating saline effluents?

The development and stability of aerobic granular sludge (AGS) was studied in two Sequencing Batch Reactors (SBRs) treating fish canning wastewater. R1 cycle comprised a fully aerobic reaction phase, while R2 cycle included a plug-flow anaerobic feeding/reaction followed by an aerobic reaction phase. The performance of the AGS reactors was compared treating the same effluents with variable salt concentrations (4.97-13.45 g NaCl/L) and organic loading rates (OLR, 1.80-6.65 kg CODs/(m3·d)). Granulation process was faster in R2 (day 34) than in R1 (day 90), however the granular biomass formed in the fully aerobic configuration was more stable to the variable feeding composition. Thus, in R1 solid retention times (SRT), up to 15.2 days, longer than in R2, up to 5.8 days, were achieved. These long SRTs values helped the retention of nitrifying organisms and provoked the increase of the nitrogen removal efficiency to 80% in R1 while it was approximately of 40% in R2. However, the presence of an anaerobic feeding/reaction phase increased the organic matter removal efficiency in R2 (80-90%) which was higher than in R1 with a fully aerobic phase (75-85%). Furthermore, in R2 glycogen-accumulating organisms (GAOs) dominated inside the granules instead of phosphorous-accumulating organisms (PAOs), suggesting that GAOs resist better the stressful conditions of a variable and high-saline influent. In terms of AGS properties an anaerobic feeding/reaction phase is not beneficial, however it enables the production of a better quality effluent.

Hybrid lattice Boltzmann method on overlapping grids.

In this work, a hybrid lattice Boltzmann method (HLBM) is proposed, where the standard lattice Boltzmann implementation based on the Bhatnagar-Gross-Krook (LBGK) approximation is combined together with an unstructured finite-volume lattice Boltzmann model. The method is constructed on an overlapping grid system, which allows the coexistence of a uniform lattice nodes spacing and a coordinate-free lattice structure. The natural adaptivity of the hybrid grid system makes the method particularly suitable to handle problems involving complex geometries. Moreover, the provided scheme ensures a high-accuracy solution near walls, given the capability of the unstructured submodel of achieving the desired level of refinement in a very flexible way. For these reasons, the HLBM represents a prospective tool for solving multiscale problems. The proposed method is here applied to the benchmark problem of a two-dimensional flow past a circular cylinder for a wide range of Reynolds numbers and its numerical performances are measured and compared with the standard LBGK ones.

A Protein Isolate from Moringa oleifera Leaves Has Hypoglycemic and Antioxidant Effects in Alloxan-Induced Diabetic Mice.

Moringa oleifera has been used in traditional medicine to treat diabetes. However, few studies have been conducted to relate its antidiabetic properties to proteins. In this study, a leaf protein isolate was obtained from M. oleifera leaves, named Mo-LPI, and the hypoglycemic and antioxidant effects on alloxan-induced diabetic mice were assessed. Mo-LPI was obtained by aqueous extraction, ammonium sulphate precipitation and dialysis. The electrophoresis profile and proteolytic hydrolysis confirmed its protein nature. Mo-LPI showed hemagglutinating activity, cross-reaction with anti-insulin antibodies and precipitation after zinc addition. Single-dose intraperitoneal (i.p.) administration of Mo-LPI (500 mg/kg·bw) reduced the blood glucose level (reductions of 34.3%, 60.9% and 66.4% after 1, 3 and 5 h, respectively). The effect of Mo-LPI was also evidenced in the repeated dose test with a 56.2% reduction in the blood glucose level on the 7th day after i.p. administration. Mo-LPI did not stimulate insulin secretion in diabetic mice. Mo-LPI was also effective in reducing the oxidative stress in diabetic mice by a decrease in malondialdehyde level and increase in catalase activity. Mo-LPI (2500 mg/kg·bw) did not cause acute toxicity to mice. Mo-LPI is a promising alternative or complementary agent to treat diabetes.

Genotyping of methicillin-resistant Staphylococcus aureus (MRSA) isolated from milk and dairy products in South Italy.

Methicillin-resistant Staphylococcus aureus (MRSA) is a pathogen emerging in hospitals as well as community and livestock. MRSA is a significant and costly public health concern because it may enter the human food chain and contaminate milk and dairy products causing foodborne illness. This study aimed to determine the occurrence and the characteristics of MRSA isolated from 3760 samples of milk and dairy products in a previous survey conducted in southern Italy during 2008-2014. Overall out of 484 S. aureus strains isolated, 40 (8.3%) were MRSA and were characterized by spa-typing, Multi-Locus Sequence Typing, SCCmec typing, Staphylococcal enterotoxins (SEs) genes, Panton-Valentine Leukocidin (PVL) genes and ability to form biofilm. The most frequently recovered STs were ST152 (t355-67.5%), followed by ST398 (t899, t108-25%), ST1 (t127-5%) and ST5 (t688-2.5%). All isolates harboured the SCCmec type V (92.5%) or IVa (25%). In one isolate (2.5%), ST398/t899, the SCCmec resulted not detected. Three isolates (7.5%) carried one or more enterotoxin encoding genes (one strain had seg, sei, sem, sen and seo genes; two strains had seh gene). The 50% of isolated strains harboured PVL-encoding genes. Molecular analysis for icaA and icaD genes showed: 72.5% icaA and icaD positive, 25% only icaD gene and one icaA and icaD negative. The detection of MRSA in food of animal origin is a potential health hazard, thus it is necessary monitoring of food-producing animals and improving hygiene standards in food practices in order to reduce the microbiological risk to minimum.

Food-Borne Viruses in Shellfish: Investigation on Norovirus and HAV Presence in Apulia (SE Italy).

Shellfish are an important vehicle for transmission of food-borne pathogens including norovirus (NoV) and hepatitis A virus (HAV). The risks related with consumption of shellfish are greater if these products are eaten raw or slightly cooked. As molluscs are filter-feeding organisms, they are able to concentrate pathogens dispersed in the water. Data on shellfish viral contamination are therefore useful to obtain a background information on the presence of contamination in the environment, chiefly in shellfish production areas and to generate a picture of the epidemiology of viral pathogens in local populations. From January 2013 to July 2015, 253 samples of bivalve molluscs collected in harvesting areas from a large coastal tract (860 km) of Southern Italy were screened for HAV and NoV of genogroups GI and GII, using real-time reverse transcription qualitative PCR. The RNA of HAV was not detected in any of the analyzed samples. In contrast, the RNA of NoV was identified in 14.2% of the samples with a higher prevalence of NoVs of genogroup GII (12.2%) than genogroup GI (1.6%). Upon sequence analysis of a short diagnostic region located in capsid region, the NoV strains were characterized as GII.2, GII.4 Sydney 2012, GII.6, GII.13, GI.4, and GI.6, all which were circulating in local populations in the same time span. These data confirm that consumption of mussels can expose consumers to relevant risks of infection. Also, matching between the NoV genotypes circulating in local population and detected in molluscs confirms the diffusion in the environment of NoVs.

"Hairy potty": Ovarian dermoid cyst with fistula to bladder / Philippine Journal of Obstetrics and Gynecology

Dermoid cysts are usually asymptomatic until complications occur. Spontaneous rupture of a dermoid into an adjacent organ is a rare complication and no such case has been reported in the Philippines. A 24-year-old primipara consulted for pilimiction. Three years earlier, she had recurrent urinary tract infection and was diagnosed to have a dermoid cyst. Left untreated, the cyst grew in size and urinary symptoms worsened. Ultrasound, CT scan and subsequent laparotomy revealed that the dermoid cyst has penetrated the bladder wall resulting to fistula formation between the dermoid and the urinary bladder. Hair and sebum were seen inside the bladder. A left salpingo-oophorectomy and partial cystectomy of the urinary bladder were done. The first locally documented case of an ovarian dermoid cyst with fistula to the bladder is presented. A review of literature is made, the predisposing factors, possible cause, diagnosis and management are discussed.

"Hairy potty": Ovarian dermoid cyst with fistula to bladder / Philippine Journal of Obstetrics and Gynecology

Dermoid cysts are usually asymptomatic until complications occur. Spontaneous rupture of a dermoid into an adjacent organ is a rare complication and no such case has been reported in the Philippines.A 24-year-old primipara consulted for pilimiction. Three years earlier, she had recurrent urinary tract infection and was diagnosed to have a dermoid cyst. Left untreated, the cyst grew in size and urinary symptoms worsened. Ultrasound, CT scan and subsequent laparotomy revealed that the dermoid cyst has penetrated the bladder wall resulting to fistula formation between the dermoid and the urinary bladder. Hair and sebum were seen inside the bladder. A left salpingo-oophorectomy and partial cystectomy of the urinary bladder were done.The first locally documented case of an ovarian dermoid cyst with fistula to the bladder is presented. A review of literature is made, the predisposing factors, possible cause, diagnosis and management are discussed.

Short communication: Isolation of Shiga toxin-producing Escherichia coli in raw milk and mozzarella cheese in southern Italy.

Shiga toxin-producing Escherichia coli (STEC) are a significant food-borne public health hazard in Europe, where most human infections are associated with 5 serogroups (O157, O26, O103, O145, and O111). In 2015, 95 food and environmental samples were examined for the presence of Shiga toxin genes (stx1 and stx2). The STEC were isolated from 2 raw milk and 1 mozzarella cheese samples that were collected in the period between June and September. To the best of our knowledge, this finding represents the first report of STEC isolation from mozzarella cheese produced in Italy, and it suggests that both the quality of raw milk used to produce mozzarella and the thermal inactivation treatment associated with the curd-stretching step should be carefully monitored.

Ischemic heart disease and early diagnosis. Study on the predictive value of 2D strain.

INTRODUCTION: Two-dimensional strain echocardiography (2D-SE) quantifies left ventricular global longitudinal strain (GLS) and global circumferential strain (GCS). Our aim was to test 2D-SE during dipyridamole stress echocardiography (Dipy-Stress) in patients with non-diagnostic result, checking by way of coronary CT angiography (CCTA) the possible presence of coronary artery disease (CAD). METHODS: Over twenty-four months 65 consecutive patients with non-diagnostic Dipy-Stress were studied by 2D-SE and by CCTA. GCS and GLS at rest and after stress were compared according to data derived from CCTA. CAD was graded as significant (stenosis ≥50%), mild (stenosis between 15 and 50%) or absent (stenosis <15%). CCTA was defined as "positive" in presence of mild CAD and "negative" in absence of stenoses. Furthermore, Δ strain was defined as follows: [(stressS-restS)/restS]×100. RESULTS: GCS at rest and after stress was similar in CCTA-positive (26±5% and 27±5% respectively) and CCTA-negative groups (27±3% and 28±3% respectively). GLS at rest was significantly reduced (P<0.0001) in CCTA-positive (23±3%) compared to CCTA-negative group (25±2%). GLS after stress was lower (P<0.0001) in CCTA-positive group (20±3%) than CCTA-negative one (26±2%). A significant reduction (P<0.0001) of GLS at rest versus after stress was found in positive-CCTA group. ΔGLS showed a significant decrease (P<0.0001) in CCTA-positive (-10±8%) compared to CCTA-negative (4.4±5.8%) group. ROC analysis of ΔGLS showed high accuracy (area under the ROC curve 0.916, 95% CI: 0.820-0.970) in distinguishing positive and negative CCTA groups. CONCLUSIONS: 2D-SE during Dipy-Stress allows, in case of non-diagnostic test, identification of mild-CAD with high sensitivity and specificity.

Start-up of two moving bed membrane bioreactors treating saline wastewater contaminated by hydrocarbons.

This work aims to assess the acclimation of microorganisms to a gradual increase of salinity and hydrocarbons, during the start-up of two moving bed membrane bioreactors (MB-MBRs) fed with saline oily wastewater. In both systems an ultrafiltration membrane was used and two types of carriers were employed: polyurethane sponge cubes (MB-MBRI) and polyethylene cylindrical carriers (MB-MBRII). A decreasing dilution factor of slops has been adopted in order to allow biomass acclimation. The simultaneous effect of salinity and hydrocarbons played an inhibitory role in biomass growth and this resulted in a decrease of the biological removal efficiencies. A reduction of bound extracellular polymeric substances and a simultaneous release of soluble microbial products (SMPs) were observed, particularly in the MB-MBRII system, probably due to the occurrence of a greater suspended biomass stress as response to the recalcitrance of substrate. On the one hand, a clear attachment of biomass occurred only in MB-MBRI and this affected the fouling deposition on the membrane surface. The processes of detachment and entrapment of biomass, from and into the carriers, significantly influenced the superficial cake deposition and its reversibility. On the other hand, in MB-MBRII, the higher production of SMPs implied a predominance of the pore blocking.

Study of aerobic granular sludge stability in a continuous-flow membrane bioreactor.

A granular continuous-flow membrane bioreactor with a novel hydrodynamic configuration was developed to evaluate the stability of aerobic granular sludge (AGS). Under continuous-flow operation (Period I), AGS rapidly lost their structural integrity resulting in loose and fluffy microbial aggregates in which filamentous bacteria were dominant. The intermittent feeding (Period II) allowed obtaining the succession of feast and famine conditions that favored the increase in AGS stability. Although no further breakage occurred, the formation of new granules was very limited, owing to the absence of the hydraulic selection pressure. These results noted the necessity to ensure, on the one hand the succession of feast/famine conditions, and on the other, the hydraulic selection pressure that allows flocculent sludge washout. This preliminary study shows that the proposed configuration could meet the first aspect; in contrast, biomass selection needs to be improved.

Pilot scale experiment with MBR operated in intermittent aeration condition: analysis of biological performance.

The effect of intermittent aeration (IA) on a MBR system was investigated. The study was aimed at analyzing different working conditions and the influence of different IA cycles on the biological performance of the MBR pilot plant, in terms of organic carbon and ammonium removal as well as extracellular polymeric substances (EPSs) production. The membrane modules were placed in a separate compartment, continuously aerated. This configuration allowed to disconnect from the filtration stage the biological phenomena occurring into the IA bioreactor. The observed results highlighted good efficiencies, in terms of organic carbon and ammonium removal. It was noticed a significant soluble microbial products (SMPs) release, likely related to the higher metabolic stress that anoxic conditions exerted on the biomass. However, the proposed configuration, with the membranes in a separate compartment, allowed to reduce the EPSs in the membrane tank even during the non-aerated phase, thus lowering fouling development.

Gait analysis in bladder exstrophy patients with and without pelvic osteotomy: a controlled experimental study.

BACKGROUND: The role of bony pelvic anomalies in bladder exstrophy is long established and has generated many papers addressing walking problems. Biomechanical studies and kinematic gait analysis were performed on very young children. AIM: A direct kinetic gait evaluation has never been performed, nor has the effect of pelvis dimorphism on the upper body been studied. DESIGN: Controlled experimental study. SETTING: Outpatients were studied at the time of periodic follow up. POPULATION: Nineteen patients with bladder exstrophy, age 14±8 years, and twenty-five healthy control participants, age 15±8 years, were enrolled in the present gait analysis study. METHODS: Clinical evaluation and standard gait analysis were performed. RESULTS: Gait analysis deviations between exstrophy patients and controls and between patients that received pelvic osteotomy (OT--6 patients) and those that did not (no-OT--13 patients) were analyzed. Bladder exstrophy significantly affects kinematics and kinetics of trunk, spine, pelvis, knee and foot; in particular: in OT, trunk retroversion, pelvic retroversion and rotation, hip adduction angle and moment, knee flexion and its maximum power during loading response increased, whereas in no-OT, spine angle, pelvic posterior tilt, hip extension, and the external rotation of the foot progression angle increased. All the kinetics parameters analyzed in the study showed lower values in the patient group than in controls. CONCLUSION: . Walking in patients with bladder exstrophy is accomplished by retroversion of the pelvis and deviations mainly in the spine angle in no-OT and in knee flexion in OT. CLINICAL REHABILITATION IMPACT: Gait analysis was shown to be an effective tool for the detection of walking deviations that should be identified early, prompting rehabilitative treatment in order to prevent spine and knee diseases.