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This work introduces the first atrial-specific in-silico human induced pluripotent stem cells-derived cardiomyocytes (hiPSC-CMs) model, based on a set of phenotype-specific I Kur ,I KCa and I K1 membrane currents. This model is built on novel in-vitro experimental data recently published by some of the co-authors to simulate the paced action potential of matured atrial-like hiPSC-CMs. The model consists of a system of stiff ordinary differential equations depending on several parameters, which have been tuned by automatic optimization techniques to closely match selected experimental biomarkers. The new model effectively simulates the electronic in-vitro hiPSC-CMs maturation process, transitioning from an unstable depolarized membrane diastolic potential to a stable hyperpolarized resting potential, and exhibits spontaneous firing activity in unpaced conditions. Moreover, our model accurately reflects the experimental rate dependence data at different cycle length and demonstrates the expected response to a specific current blocker. This atrial-specific in-silico model provides a novel computational tool for electrophysiological studies of cardiac stem cells and their applications to drug evaluation and atrial fibrillation treatment., Competing Interests: Declaration of competing interest There are no conflicts of interest: None Declared., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: Xylella fastidiosa is an important causative agent of Olive Quick Decline Syndrome in the Apulia region of Italy. The current study evaluated the bioefficacy of three entomopathogenic fungal strains: Beauveria bassiana SGB7004, Metarhizium robertsii SGB1K, and Akanthomyces lecanii SGB4711 against Philaenus spumarius the main vector of this pathogen, under laboratory conditions. Pathogenicity bioassays were performed by dipping nymphs and adults of P. spumarius in an aqueous suspension of powdered fungal culture (PFC) or conidial suspension (CS) of the three fungal strains., Results: Both B. bassiana SGB7004 and M. robertsii SGB1K affected the viability of nymphs, resulting in more than 80% mortality at 48 h post treatment, while the effect of A. lecanii SGB4711 was not statistically significant. On adults, all three biocontrol strains were effective in a time- and concentration-dependent manner. The PFCs of B. bassiana SGB7004, M. robertsii SGB1K, and A. lecanii SGB4711 at the highest concentration tested (120 mg mL -1 ) resulted in 97%, 83% and 27% mortality at the trial endpoint (120 h), respectively. Mycelial growth was observed on 38.5%, 37.0% and 61.5% of dead insects treated with B. bassiana SGB7004 (2.3 × 10 8  CFU mL -1 ), M. robertsii SGB1K (3.8 × 10 6  CFU mL -1 ) and A. lecanii SGB4711 (5.4 × 10 8  CFU mL -1 ), respectively. None of the PFCs of the tested strains was pathogenic when injected into nymph spittle., Conclusions: Beauveria bassiana SGB7004 and M. robertsii SGB1K significantly affected the survival of P. spumarius nymphs and adults, while A. lecanii SGB4711 was not effective on nymphs and only slightly effective against adults. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Accumulating senescent cells within tissues contribute to the progression of aging and age-related diseases. Botanical extracts, rich in phytoconstituents, present a useful resource for discovering therapies that could target senescence and thus improve healthspan. Here, we show that daily oral administration of a standardized extract of Salvia haenkei (Haenkenium (HK)) extended lifespan and healthspan of naturally aged mice. HK treatment inhibited age-induced inflammation, fibrosis and senescence markers across several tissues, as well as increased muscle strength and fur thickness compared with age-matched controls. We also found that HK treatment reduced acutely induced senescence by the chemotherapeutic agent doxorubicin, using p16 LUC reporter mice. We profiled the constituent components of HK by mass spectrometry, and identified luteolin-the most concentrated flavonoid in HK-as a senomorphic compound. Mechanistically, by performing surface plasmon resonance and in situ proximity ligation assay, we found that luteolin disrupted the p16-CDK6 interaction. This work demonstrates that administration of HK promotes longevity in mice, possibly by modulating cellular senescence and by disrupting the p16-CDK6 interaction., (© 2024. The Author(s).)

(1) This study evaluates the impact of the CT-guided SIRIO augmented reality navigation system on the procedural efficacy and clinical outcomes of neuroprotection in vertebral thermal ablation (RTA) for primary and metastatic bone tumors. (2) Methods: A retrospective non-randomized analysis of 28 vertebral RTA procedures was conducted, comparing 12 SIRIO-assisted and 16 non-SIRIO-assisted procedures. The primary outcomes included dose-length product (DLP) and epidural dissection time. The secondary outcomes included technical success, complication rates, and pain scores at procedural time (VAS Time 0) and three months post-procedure (VAS Time 1). The statistical analyses included t-tests, Mann-Whitney U tests, and multiple regression. (3) Results: SIRIO-assisted procedures significantly reduced DLP (307.42 mGycm vs. 460.31 mGycm, p = 2.23 × 10 -8 ) and procedural epidural dissection time (13.48 min vs. 32.26 min, p = 2.61 × 10 -12 ) compared to non-SIRIO-assisted procedures. Multiple regression confirmed these reductions were significant (DLP: β = -162.38, p < 0.001; time: β = -18.25, p < 0.001). Pain scores (VAS Time 1) did not differ significantly between groups, and tumor type did not significantly influence outcomes. (4) Conclusions: The SIRIO system enhances neuroprotection efficacy and safety, reducing radiation dose and procedural time during spine tumoral ablation while maintaining consistent pain management outcomes.

Endophytic fungi live inside virtually every plant species, without causing any apparent disease or damage to the host. Nevertheless, under particular conditions, mutualistic lifestyle of endophytes may change to pathogenic. In this study, the biodiversity of Alternaria and Fusarium species, the two most abundant endophytic fungi isolated from healthy potato plants in two climatically different regions of Iran, Ardebil in the north-west and Kerman in the south-east, was investigated. Seventy-five Fusarium strains and 83 Alternaria strains were molecularly characterized by multi-locus gene sequencing. Alternaria strains were characterized by the sequences of gpd and caM gene fragments and the phylogenetic tree was resolved in 3 well-separated clades. Seventy-three strains were included in the clade A, referred as Alternaria section, 6 strains were included in clade B, referred as Ulocladioides section, and 4 strains were included in clade C, referred as Infectoriae section. Fusarium strains, identified by sequencing the translation elongation factor 1α ( tef1 ), β-tubulin ( tub2 ) and internal transcribed spacer (ITS) genomic regions, were assigned to 13 species, viz. F. brachygibosum , F. clavum , F. equiseti , F. flocciferum , F. incarnatum , F. nirenbergiae , F. nygamai , F. oxysporum , F. proliferatum , F. redolens , F. sambucinum , F. solani and F. thapsinum . Twenty-six selected strains, representative of F. equiseti , F. nirenbergiae , F. oxysporum , F. nygamai , F. proliferatum , and F. sambucinum , were also tested for production of the mycotoxins deoxynivalenol (DON), nivalenol (NIV), diacetoxyscirpenol (DAS), T-2 toxin (T-2), beauvericin (BEA), enniatins (ENNs), fumonisins (FBs), fusaric acid (FA) and moniliformin (MON). None of the tested strains produced trichothecene toxins (DON, NIV, DAS and T-2). Two out of 2  F. equiseti isolates, 1/6  F. oxysporum , 1/3  F. proliferatum , and 1/9  F. nygamai did not produce any of the tested toxins; the rest of strains produced one or more BEA, ENNs, FBs, FA and MON toxins. The most toxigenic strain, F. nygamai ITEM-19012, produced the highest quantities of FBs (7946, 4693 and 4333 μg/g of B1, B2, and B3 respectively), along with the highest quantities of both BEA (4190 μg/g) and MON (538 μg/g). These findings suggest that contamination of potato tubers with mycotoxins in the field or at post-harvest, due to a change in lifestyle of endophytic microflora, should be carefully considered and furtherly investigated., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Miriam Haidukowski reports travel was provided by Iran National Science Foundation. Miriam Haidukowski reports financial support, administrative support, article publishing charges, and equipment, drugs, or supplies were provided by Institute of Sciences of Food Production, National Research Council, Italy. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

Background: Laparoscopic cholecystectomy is the most common minimally invasive abdominal surgery procedure performed in Western countries; it offers several advantages over laparotomy but still carries some risks, such as intraoperative spillage of bile and gallstones. Diagnosis of dropped gallstones could be challenging, it is frequently delayed, and this can lead to further complications such as abscesses formation., Methods: We report the history of a 51-year-old male with persistent dull abdominal pain in association to appetite loss, vomiting episodes and changes in regular bowel habits, a past medical history of laparoscopic cholecystectomy for biliary lithiasis (1.5 years earlier) and minimum elevation of inflammatory markers and gamma-GT values., Results: Ultrasound examination showed perihepatic stones and magnetic resonance imaging revealed the presence of multiple perihepatic abscesses, findings compatible with fibrotic-inflammatory phenomena from 'dropped gallstones'. A re-laparoscopy was then performed with an abscess collection containing multiple gallstones; a liver wedge resection was also required due to strong adhesions. At follow up, the patient had improved both on clinical and biochemical perspective., Conclusion: Dropped gallstones are an underreported cause of morbidity and diagnostic dilemmas in subjects who underwent to laparoscopic cholecystectomy, in relation to infectious complications that can occur even several months or years after surgery. Imaging represents a valuable aid in the correct non-invasive diagnostic process, but proper awareness of this insidious condition is necessary. Surgeons and radiologists should always consider this eventuality in the differential diagnosis of a patient presenting with abdominal abscesses and history of cholecystectomy.

Purpose: To assess the effectiveness and safety of cryoablation (CRA) for the treatment of recurrent or oligometastatic solid tumors located in the thoracoabdominal soft tissues., Materials and Methods: Twenty-two percutaneous CRA procedures performed in 19 patients to treat recurrent or oligometastatic tumors in thoracoabdominal soft tissue were retrospectively examined. All procedures were performed between January 2015 and June 2021 under ultrasound and computed tomography (CT) guidance, and the most complex procedures were performed with CT-based navigation systems. The histology of the primary tumors included colorectal adenocarcinoma, squamous cell lung carcinoma, pancreatic adenocarcinoma, renal cell carcinoma, and hepatocellular carcinoma. Adverse events, technical success, and local tumor control were analyzed., Results: The mean age of the patients was 66.5 years, with a mean tumor size of 24.8 mm. The mean time of the procedures was 68 minutes, with a mean number of 2.5 cryoprobes used. Hydrodissection was performed in 63% of the procedures to protect the surrounding anatomical structures. The mean size of the ice ball, measured on axial CT scans at the end of the procedures, was 43.5 mm. No severe adverse events were observed. Technical success was achieved in all cases. Three patients experienced local tumor progression (2 residual disease and 1 recurrence), which were successfully treated with a second CRA procedure., Conclusions: Percutaneous CRA is a safe and effective therapy in selected cases of recurrent or oligometastatic tumors in the thoracoabdominal soft tissues., (Copyright © 2023 SIR. Published by Elsevier Inc. All rights reserved.)

In a judiciary setting, questions regarding the mechanisms of transfer, persistence, and recovery of DNA are increasingly more common. The forensic expert is now asked to evaluate the strength of DNA trace evidence at activity level, thus assessing if a trace, given its qualitative and quantitative features, could be the result of an alleged activity. The present study is the reproduction of a real-life casework scenario of illicit credit card use by a co-worker (POI) of its owner (O). After assessing the shedding propensity of the participants, differences in DNA traces' qualitative and quantitative characteristics, given scenarios of primary and secondary transfer of touch DNA on a credit card, a non-porous plastic support, were investigated. A case-specific Bayesian Network to aid statistical evaluation was created and discrete observations, meaning the presence/absence of POI as a major contributor in both traces from direct and secondary transfer, were used to inform the probabilities of disputed activity events. Likelihood Ratios at activity level (LRα) were calculated for each possible outcome resulting from the DNA analysis. In instances where only POI and POI plus an unknown individual are retrieved, the values obtained show moderate to low support in favour of the prosecution proposition.

Human induced pluripotent stem cell (hiPSC)-derived cardiomyocytes (CM) constitute a mixed population of ventricular-, atrial-, nodal-like cells, limiting the reliability for studying chamber-specific disease mechanisms. Previous studies characterised CM phenotype based on action potential (AP) morphology, but the classification criteria were still undefined. Our aim was to use in silico models to develop an automated approach for discriminating the electrophysiological differences between hiPSC-CM. We propose the dynamic clamp (DC) technique with the injection of a specific I K1 current as a tool for deriving nine electrical biomarkers and blindly classifying differentiated CM. An unsupervised learning algorithm was applied to discriminate CM phenotypes and principal component analysis was used to visualise cell clustering. Pharmacological validation was performed by specific ion channel blocker and receptor agonist. The proposed approach improves the translational relevance of the hiPSC-CM model for studying mechanisms underlying inherited or acquired atrial arrhythmias in human CM, and for screening anti-arrhythmic agents., (© 2023. The Author(s).)

Background: The meadow spittlebug Philaenus spumarius L. is the vector for the bacterium Xylella fastidiosa subspecies pauca, involved in olive quick decline syndrome (OQDS) in Salento (Italy). Control of P. spumarius is key to limiting transmission of the bacterium, and an innovative approach can be based on effective natural compounds and biocontrol agents. Entomopathogenic fungi are an important source of bioactive natural molecules that play a role in the relationship between microorganisms and insects., Results: Pathogenicity bioassays, performed by dipping adults of P. spumarius in either fungal culture suspension (120 mg mL -1 ) or cell-free culture supernatant of Trichoderma chlorosporum GJS 91-150, showed, respectively, 97% and 87% death within 24 h. The effect was dose-dependent. In laboratory bioassays, the powdered fungal culture of T. chlorosporum GJS 91-150 did not exhibit pathogenic activity when injected into nymph spittle., Conclusions: T. chlorosporum GJS 91-150 affected the survival of P. spumarius adults. The lethal effect was not associated with the development of mycelium on the cuticle, but seems due, at least partly, to fungal metabolites released in the culture medium. The fungus tested here has good potential for the development of effective low-environmental impact control strategies for P. spumarius and suppression of X. fastidiosa. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Background: sequential or simultaneous applications of multiple antennas have been proposed to create larger ablation zone; however, there is a lack of data in patients affected by liver tumors, with potentially different results from animal liver models. The purpose of this study was to evaluate efficacy and safety of liver percutaneous microwave ablation using simultaneous activation of two antennas to treat lesions bigger than 2,5 cm; particularly the focus was assessing whether the ratio of ablation zone volume in millimeters to applied energy in kilojoules [R(AZ:E)] differs between hepatocellular carcinoma in a cirrhotic liver and liver metastasis and if it is correlated to complications incidence or recurrence of disease., Methods: Fifty-five liver microwave ablation performed with two simultaneous antennas from March 2017 to June 2021 were retrospectively reviewed; 9 procedures were excluded due to the association with Chemoembolization. Size, shape, volume of lesions and ablation zones were recorded. Technical success was defined as complete devascularization of the treated area at the post-procedural CT. R(AZ:E) was determined dividing the ablation zone volume in mm3 by the amount of energy in kilojoules applied in each procedure and complications were reported., Results: Technical success was achieved in all the procedures. Mean R(AZ:E) was 0,75 ± 0,58. T-student test for patients with HCC and patients with metastasis about R(AZ:E) was significant ( p  = 0.03). The incidence of bilomas was lower for HCC ( p  = 0.022). One-month follow-up showed Complete Response (CR) in 44/46 (95,6%) patients; Three-six months follow-up demonstrated: CR in 43/46 (93.5%) cases and 12 months follow-up highlighted CR in 40/45 (88,9%) cases., Conclusions: These results provide preliminary evidence of efficacy and safety of simultaneous liver MWA using two antennas, highlighting the importance of procedural indications.

We present the case of a 50-year-old woman affected by a rectal gastrointestinal stromal tumor (GIST), with a recurrence in pre-sacral and pre-coccygeal space after surgery and Imatinib therapy. GISTs are the most common mesenchymal tumors of the gastrointestinal tract and rectal GISTs are rare (only 2% of cases); magnetic resonance and computed tomography are the main imaging techniques for diagnosis and follow-up, while ultrasound and contrast-enhanced ultrasound may be useful to perform a percutaneous biopsy, as in the case presented: the imaging features of the lesion in all these imaging methods are displayed., (© 2022 The Authors. Published by Elsevier Inc. on behalf of University of Washington.)

Marantic endocarditis is a rare entity often diagnosed in the setting of advanced malignant disease. We present the case of a 66-year-old man with stage IVb non-small cell lung cancer, with large tricuspid and small aortic valve vegetations. A multidisciplinary team determined the patient to be a poor surgical candidate, but given the likelihood of pulmonary and systemic embolization, an intervention was required. We proposed AngioVac-assisted vegetation debulking of the tricuspid valve to be the best approach in consideration of the small aortic valve vegetation and lung cancer. He successfully underwent this procedure and was discharged on postprocedure day 2., (© 2022 The Authors.)

Aflatoxins, which are produced mainly by Aspergillus flavus and A. parasiticus , are recognized as the most toxic mycotoxins, which are strongly carcinogenic and pose a serious threat to human and animal health. Therefore, strategies to degrade or eliminate aflatoxins in agro-products are urgently needed. We investigated 65 Trichoderma isolates belonging to 23 species for their aflatoxin B 1 (AFB 1 )-degrading capabilities. Trichoderma reesei CGMCC3.5218 had the best performance, and degraded 100% of 50 ng/kg AFB 1 within 3 days and 87.6% of 10 μg/kg AFB 1 within 5 days in a liquid-medium system. CGMCC3.5218 degraded more than 85.0% of total aflatoxins (aflatoxin B 1 , B 2 , G 1 , and G 2 ) at 108.2-2323.5 ng/kg in artificially and naturally contaminated peanut, maize, and feed within 7 days. Box-Behnken design and response surface methodology showed that the optimal degradation conditions for CGMCC3.5218 were pH 6.7 and 31.3°C for 5.1 days in liquid medium. Possible functional detoxification components were analyzed, indicating that the culture supernatant of CGMCC3.5218 could efficiently degrade AFB 1 (500 ng/kg) with a ratio of 91.8%, compared with 19.5 and 8.9% by intracellular components and mycelial adsorption, respectively. The aflatoxin-degrading activity of the fermentation supernatant was sensitive to proteinase K and proteinase K plus sodium dodecyl sulfonate, but was stable at high temperatures, suggesting that thermostable enzymes or proteins in the fermentation supernatant played a major role in AFB 1 degradation. Furthermore, toxicological experiments by a micronucleus assay in mouse bone marrow erythrocytes and by intraperitoneal injection and skin irritation tests in mice proved that the degradation products by CGMCC3.5218 were nontoxic. To the best of our knowledge, this is the first comprehensive study on Trichoderma aflatoxin detoxification, and the candidate strain T. reesei CGMCC3.5218 has high efficient and environment-friendly characteristics, and qualifies as a potential biological detoxifier for application in aflatoxin removal from contaminated feeds., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be constructed as a potential conflict of interest., (Copyright © 2022 Yue, Ren, Fu, Wei, Altomare, Haidukowski, Logrieco, Zhang and Li.)

The coumarin core (i.e., 1-benzopyran-2 (2 H )-one) is a structural motif highly recurrent in both natural products and bioactive molecules. Indeed, depending on the substituents and branching positions around the byciclic core, coumarin-containing compounds have shown diverse pharmacological activities, ranging from anticoagulant activities to anti-inflammatory, antimicrobial, anti-HIV and antitumor effects. In this survey, we have reported the main scientific results of the 20-years investigation on the coumarin core, exploited by the research group headed by Prof. Angelo Carotti (Bari, Italy) either as a scaffold or a pharmacophore moiety in designing novel biologically active small molecules., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Pisani, Catto, Muncipinto, Nicolotti, Carrieri, Rullo, Stefanachi, Leonetti and Altomare.)

The antibiotic and nematocidal activities of extracts from two coastal lichen species collected on Lampedusa Island (Sicily), Ramalina implexa Nyl. and Roccella phycopsis Ach., were tested. Methyl orsellinate, orcinol, (+)-montagnetol, and for the first time 4-chlororcinol were isolated from Roccella phycopsis . (+)-Usnic acid was obtained from Ramalina implexa . The crude organic extract of both lichen species showed strong antibiotic activity against some bacterial species and nematocidal activity. Among all the pure metabolites tested against the infective juveniles (J2) of the root-knot nematode (RKN) Meloydogine incognita , (+)-usnic acid, orcinol, and (+)-montagnetol had significant nematocidal activity, comparable with that of the commercial nematocide Velum ® Prime, and thus they showed potential application in agriculture as a biopesticide. On the contrary, methyl orsellinate and 4-chlororcinol had no nematocidal effect. These results suggest that the substituent pattern at ortho -para-position in respect to both hydroxyl groups of resorcine moiety, which is present in all metabolites, seems very important for nematocidal activity. The organic extracts of both lichens were also tested against some Gram-positive and Gram-negative bacteria. Both extracts were active against Gram-positive species. The extract of Ramalina implexa showed, among Gram-negative species, activity against Escherichia coli and Acinetobacter baumannii , while that from Roccella phycopsis was effective towards all test strains, with the exception of Pseudomonas aeruginosa. The antimicrobial activity of (+)-usnic acid, methyl orsellinate, and (+)-montagnetol is already known, so tests were focused on orcinol and 4-chlororcinol. The former showed antibacterial activity against all Gram positive and Gram-negative test strains, with the exception of A. baumannii and K. pneumoniae , while the latter exhibited a potent antibacterial activity against Gram-positive test strains and among Gram-negative strains, was effective against A. baumannii and K. pneumonia . These results suggest, for orcinol and 4-chlororcinol, an interesting antibiotic potential against both Gram-positive and Gram-negative bacterial strains.

Rationale: Aging in the heart is a gradual process, involving continuous changes in cardiovascular cells, including cardiomyocytes (CMs), namely cellular senescence. These changes finally lead to adverse organ remodeling and resulting in heart failure. This study exploits CMs from human induced pluripotent stem cells (iCMs) as a tool to model and characterize mechanisms involved in aging. Methods and Results: Human somatic cells were reprogrammed into human induced pluripotent stem cells and subsequently differentiated in iCMs. A senescent-like phenotype (SenCMs) was induced by short exposure (3 hours) to doxorubicin (Dox) at the sub-lethal concentration of 0.2 µM. Dox treatment induced expression of cyclin-dependent kinase inhibitors p21 and p16, and increased positivity to senescence-associated beta-galactosidase when compared to untreated iCMs. SenCMs showed increased oxidative stress, alteration in mitochondrial morphology and depolarized mitochondrial membrane potential, which resulted in decreased ATP production. Functionally, when compared to iCMs, SenCMs showed, prolonged multicellular QTc and single cell APD, with increased APD variability and delayed afterdepolarizations (DADs) incidence, two well-known arrhythmogenic indexes. These effects were largely ascribable to augmented late sodium current (I NaL ) and reduced delayed rectifier potassium current (Ikr). Moreover sarcoplasmic reticulum (SR) Ca 2+ content was reduced because of downregulated SERCA2 and increased RyR2-mediated Ca 2+ leak. Electrical and intracellular Ca 2+ alterations were mostly justified by increased CaMKII activity in SenCMs. Finally, SenCMs phenotype was furtherly confirmed by analyzing physiological aging in CMs isolated from old mice in comparison to young ones. Conclusions: Overall, we showed that SenCMs recapitulate the phenotype of aged primary CMs in terms of senescence markers, electrical and Ca 2+ handling properties and metabolic features. Thus, Dox-induced SenCMs can be considered a novel in vitro platform to study aging mechanisms and to envision cardiac specific anti-aging approach in humans., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Purpose: To evaluate efficacy and safety of radiofrequency ablation (RFA) and microwave ablation (MWA) in unresectable lung malignancies., Methods: Data regarding patients with primary and secondary lung tumors treated with RFA or MWA from 2008 to 2020 were reviewed retrospectively. Primary study objectives such as technical success, primary and secondary technique efficacy rates, local tumor progression (LTP) rate, LPT-free survival (LPTFS) and overall survival (OS) were assessed. Secondary study objectives were side effects and complications. RFA and MWA were compared using the Chi-square test for continuous variables. Kaplan-Meier curves were calculated for survival statistical analysis., Results: A total of 113 patients with primary or secondary lung tumor underwent 74 RFA (48%) and 81 MWA (52%). Technical success rate was 151/155 (97%); primary and secondary technique efficacy rates were 123/155 (79%) and 129/155 (83%), respectively. During the entire study follow-up, 32 cases experienced disease progression (20%), of which 18 underwent repeat ablation (12%), in 6 cases with success (4%). Residual unablated tumor happened in 4/155 cases (3%). LTP occurred in 28/155 cases (17%). The only factor associated with poorer LTP-FS was lesion diameter ≥ 30 mm (P < 0.05). One-, 3- and 5-years LTP-FS was 83%, 82%, 82%, respectively. One-, 3- and 5-years OS of the entire population was 87%, 74%, 73%, respectively. Minor and major complication rates were 53/155 (34%) and 29/155 (19%), respectively., Conclusions: In conclusion, this study confirms the appropriateness of RFA and MWA for lung tumors treatment, in terms of safety and efficacy., (© 2022. Italian Society of Medical Radiology.)

Aims: Diabetic cardiomyopathy is a multifactorial disease characterized by an early onset of diastolic dysfunction (DD) that precedes the development of systolic impairment. Mechanisms that can restore cardiac relaxation improving intracellular Ca2+ dynamics represent a promising therapeutic approach for cardiovascular diseases associated to DD. Istaroxime has the dual properties to accelerate Ca2+ uptake into sarcoplasmic reticulum (SR) through the SR Ca2+ pump (SERCA2a) stimulation and to inhibit Na+/K+ ATPase (NKA). This project aims to characterize istaroxime effects at a concentration (100 nmol/L) marginally affecting NKA, in order to highlight its effects dependent on the stimulation of SERCA2a in an animal model of mild diabetes., Methods and Results: Streptozotocin (STZ) treated diabetic rats were studied at 9  weeks after STZ injection in comparison to controls (CTR). Istaroxime effects were evaluated in vivo and in left ventricular (LV) preparations. STZ animals showed (i) marked DD not associated to cardiac fibrosis, (ii) LV mass reduction associated to reduced LV cell dimension and T-tubules loss, (iii) reduced LV SERCA2 protein level and activity and (iv) slower SR Ca2+ uptake rate, (v) LV action potential (AP) prolongation and increased short-term variability (STV) of AP duration, (vi) increased diastolic Ca2+, and (vii) unaltered SR Ca2+ content and stability in intact cells. Acute istaroxime infusion (0.11 mg/kg/min for 15 min) reduced DD in STZ rats. Accordingly, in STZ myocytes istaroxime (100 nmol/L) stimulated SERCA2a activity and blunted STZ-induced abnormalities in LV Ca2+ dynamics. In CTR myocytes, istaroxime increased diastolic Ca2+ level due to NKA blockade albeit minimal, while its effects on SERCA2a were almost absent., Conclusions: SERCA2a stimulation by istaroxime improved STZ-induced DD and intracellular Ca2+ handling anomalies. Thus, SERCA2a stimulation can be considered a promising therapeutic approach for DD treatment., (© The Author(s) 2021. Published by Oxford University Press on behalf of the European Society of Cardiology.)

Background/aim: The prediction of a sub-areolar tissue infiltration in breast cancer (BC) patients could be helpful in selecting the best functional outcome according to several reconstructive oncoplastic or radical techniques. This study aims to evaluate the diagnostic performance of preoperative ultrasound (US) guided sub-areolar biopsy (SAB) in detecting occult nipple involvement, in comparison with the definitive pathological examination of tissue after surgery., Patients and Methods: We prospectively recorded clinical and pathological data of 46 consecutive patients scheduled for breast conserving surgery or nipple-areola sparing mastectomy. All cases underwent preoperative SAB and the results were compared with the histopathology of the dissected tumors and their biological characteristics. All parameters were correlated with nipple involvement by univariate and multivariate analysis., Results: The sensitivity, specificity, overall accuracy, positive predictive value, and negative predictive value of the SAB examination for nipple staging were 60%, 97.5%, 75% and 95.2%, respectively. According to the clinicopathological features, the bivariate analysis did not show a significant interaction between patient age, tumor size or type, lymph node status, lymphatic vascular invasion, histologic grade, ER, PR, Ki-67 status, HER2 amplification, multifocal or multicentric disease and positive NAC assessment (p>0.05)., Conclusion: A preoperative sample of retroareolar tissue obtained by US-guided biopsy could be considered a reproducible, mini-invasive diagnostic procedure useful to facilitate immediate breast reconstruction with implants or through conservative oncoplastic approaches, thereby guiding clinical practice., (Copyright© 2022, International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background: Augmented reality navigation system for percutaneous computed tomography (CT)-guided pulmonary biopsies has recently been introduced. There are no studies in literature about its use for ground glass lesions biopsies. The aim of this study is to evaluate the effectiveness of an augmented reality infrared navigation system performance on CT-guided percutaneous lung ground glass opacity (GGO) biopsy compared to a standard CT-guided technique., Methods: A total of 80 patients with lung GGO who underwent to a percutaneous CT-guided lung biopsy with an augmented reality infrared navigation system were retrospectively enrolled in the study. Comparison was performed with a group of 80 patients who underwent to lung biopsy with the standard CT-guided technique. Evaluation of maximum lesion diameter (MLD), distance between lesion and pleural surface (DPS), distance travelled by the needle (DTP), procedural time, validity of histological sample, procedural complications and the radiation dose to the patient's chest were recorded for each patient of both groups. In addition, each group was divided into two subgroups based on lesion size, according to a cut-off of 1.5 cm (<1.5 cm; ≥1.5 cm)., Results: Augmented reality navigation system showed a significant reduction in procedural time, radiation dose administrated to patients and complications rate compared to a standard CT-guided technique. Technical success was achieved in the 100% of cases in both groups, but the diagnostical success was higher in the group where patients underwent to lung biopsies with the use of navigation system. We also found that using an augmented reality navigation system increases the diagnostical success rate for lesion <1.5 cm. MLD, DPS and DTP did not differ significantly between the two groups of patients., Conclusions: The use of an augmented reality navigation system for percutaneous CT-guided pulmonary GGO biopsies has demonstrated a lower incidence of post-procedural complications, a significantly reduction of the radiation dose administered to patients and a higher diagnostical success rate., Competing Interests: Conflicts of Interest: All authors have completed the ICMJE uniform disclosure form (available at https://jtd.amegroups.com/article/view/10.21037/jtd-21-1285/coif). PC serves as an unpaid editorial board member of Journal of Thoracic Disease from November 2016 to October 2022. The other authors have no conflicts of interest to declare., (2022 Journal of Thoracic Disease. All rights reserved.)

The inhibitory action of 20 antagonistic Trichoderma isolates against the aflatoxigenic isolate A. flavus ITEM 9 (Af-9) and their efficacy in reducing aflatoxin formation in vitro were examined. Production of metabolites with inhibitory effect by the Trichoderma isolates was also investigated. Antagonistic effect against Af-9 was assessed by inhibition of radial growth of the colonies and by fungal interactions in dual confrontation tests. A total of 8 out of 20 isolates resulted in a significant growth inhibition of 3-day-old cultures of Af-9, ranging from 13% to 65%. A total of 14 isolates reduced significantly the aflatoxin B 1 (AfB 1 ) content of 15-day-old Af-9 cultures; 4 were ineffective, and 2 increased AfB 1 . Reduction of AfB 1 content was up to 84.9% and 71.1% in 7- and 15-day-old cultures, respectively. Since the inhibition of Af-9 growth by metabolites of Trichoderma was not necessarily associated with inhibition of AfB 1 production and vice versa, we investigated the mechanism of reduction of AfB 1 content at the molecular level by examining two strains: one (T60) that reduced both growth and mycotoxin content; and the other (T44) that reduced mycotoxin content but not Af-9 growth. The expression analyses for the two regulatory genes aflR and aflS , and the structural genes aflA , aflD , aflO and aflQ of the aflatoxin biosynthesis cluster indicated that neither strain was able to downregulate the aflatoxin synthesis, leading to the conclusion that the AfB 1 content reduction by these Trichoderma strains was based on other mechanisms, such as enzyme degradation or complexation. Although further studies are envisaged to identify the metabolites involved in the biocontrol of A. flavus and prevention of aflatoxin accumulation, as well as for assessment of the efficacy under controlled and field conditions, Trichoderma spp. qualify as promising agents and possible alternative options to other biocontrol agents already in use.

Development of more sensitive imaging techniques has caused an increase in the number of diagnosed small renal tumors. Approximately 2-3% of these lesions are proved to be angiomyolipomas (AML), a rare benign tumor of the kidney sometimes causing pain and hematuria. The most required approach is observation, but in the case of recurrent symptoms or larger tumors, which may cause bleeding, a more active treatment is required. We present two cases of symptomatic AML tumors of different sizes in the kidney: one treated with transarterial embolization (TAE), and the other with percutaneous cryoablation (CRA). The lesions were diagnosed on the basis of contrast-enhanced computed tomography (CT) scan and magnetic resonance imaging (MRI). Both treatments proved to be effective and safe for treating renal AMLs. A follow-up carried out, based on contrast-enhanced CT scan, confirmed complete treatment of AML and decreased lesion size. There are myriad minimally invasive approaches for the treatment of renal AMLs, and the preservation of renal function remains a priority. The most popular treatment option is the selective renal artery embolization. Owing to its limited invasiveness, CRA could be an attractive option for the preventive treatment of AML., Competing Interests: This study was not supported by any funding. The authors declare that no conflict of interest exists., (Copyright: Pacella G, et al.)

(1) Background: The aim of this retrospective study is to assess safety and efficacy of lung radiofrequency (RFA) and microwave ablation (MWA) using an augmented reality computed tomography (CT) navigation system (SIRIO) and to compare it with the standard CT-guided technique. (2) Methods: Lung RFA and MWA were performed with an augmented reality CT 3D navigation system (SIRIO) in 52 patients. A comparison was then performed with a group of 49 patients undergoing the standard CT-guided technique. All the procedures were divided into four groups based on the lesion diameter (>2 cm or ≤2 cm), and procedural time, the number of CT scans, radiation dose administered, and complications rate were evaluated. Technical success was defined as the presence of a "ground glass" area completely covering the target lesion at the immediate post-procedural CT. (3) Results: Full technical success was achieved in all treated malignant lesions for all the considered groups. SIRIO-guided lung thermo-ablations (LTA) displayed a significant decrease in the number of CT scans, procedure time, and patients' radiation exposure ( p < 0.001). This also resulted in a dosage reduction in hypnotics and opioids administrated for sedation during LTA. No significant differences were observed between the SIRIO and non-SIRIO group in terms of complications incidence. (4) Conclusions: SIRIO is an efficient tool to perform CT-guided LTA, displaying a significant reduction ( p < 0.001) in the number of required CT scans, procedure time, and patients' radiation exposure.

(1) Background: The purpose of this study is to evaluate the impact of an augmented reality navigation system (SIRIO) for percutaneous biopsies and ablative treatments on bone lesions, compared to a standard CT-guided technique. (2) Methods: Bioptic and ablative procedures on bone lesions were retrospectively analyzed. All procedures were divided into SIRIO and Non-SIRIO groups and in <2 cm and >2 cm groups. Number of CT-scans, procedural time and patient's radiation dose were reported for each group. Diagnostic accuracy was obtained for bioptic procedures. (3) Results: One-hundred-ninety-three procedures were evaluated: 142 biopsies and 51 ablations. Seventy-four biopsy procedures were performed using SIRIO and 68 under standard CT-guidance; 27 ablative procedures were performed using SIRIO and 24 under standard CT-guidance. A statistically significant reduction in the number of CT-scans, procedural time and radiation dose was observed for percutaneous procedures performed using SIRIO, in both <2 cm and >2 cm groups. The greatest difference in all variables examined was found for procedures performed on lesions <2 cm. Higher diagnostic accuracy was found for all SIRIO-assisted biopsies. No major or minor complications occurred in any procedures. (4) Conclusions: The use of SIRIO significantly reduces the number of CT-scans, procedural time and patient's radiation dose in CT-guided percutaneous bone procedures, particularly for lesions <2 cm. An improvement in diagnostic accuracy was also achieved in SIRIO-assisted biopsies.

Background/aim: Prior studies have underlined the prognostic relevance of pathological complete response (pCR) after neoadjuvant chemotherapy (NAC) in breast cancer. However, an accurate demonstration of treatment efficacy is dependent on its potential to predict long-term outcomes of recurrence and death, and this issue remains somewhat controversial., Patients and Methods: One hundred and sixty-nine patients with breast cancer (BC) treated with NAC followed by surgery were enrolled in this retrospective study. After carrying out multivariable analyses, involving baseline characteristics (tumor stage, nodal status, histological grade, biological profile) and response status, we analysed the association between pCR and disease-free (DFS) and overall survival (OS) in various subtypes. Moreover, we investigated several residual disease-scoring combinations to check whether they could discriminate prognostic subsets according to their variable tumor range after NAC., Results: Overall, factors associated with pCR were non-luminal subtype (p<0.001), high grade (p=0.001) and HER2-overexpression (p=0.001). Residual tumor and nodal stage after NAC significantly correlated with DFS (p=0.007) and OS (p<0.001). Similarly, pCR after NAC showed significantly better DFS (p=0.01), particularly for HER2-positive (p=0.003), triple-negative (p=0.019) and HER2-positive Luminal B profiles (p=0.019). However, there was no statistical difference in the OS among patients who had PCR, compared to absence of pCR (p=0.40)., Conclusion: Extent of residual disease and evidence of regression provide helpful prognostic details in BC patients treated with NAC. Achieving pCR after NAC is related with significantly better DFS, with the potential of maximized breast and axillary conservation based on clinical response. The distribution of expertise in a cross-disciplinary setting could provide safe and favourable prognosis, while improving cosmetic outcomes and quality of life., (Copyright © 2021 International Institute of Anticancer Research (Dr. George J. Delinasios), All rights reserved.)

Background: Combined treatment with anthracyclines (e.g., doxorubicin; Dox) and trastuzumab (Trz), a humanized anti-human epidermal growth factor receptor 2 (HER2; ErbB2) antibody, in patients with HER2-positive cancer is limited by cardiotoxicity, as manifested by contractile dysfunction and arrhythmia. The respective roles of the two agents in the cardiotoxicity of the combined therapy are incompletely understood., Objective: To assess cardiac performance, T-tubule organization, electrophysiological changes and intracellular Ca 2+ handling in cardiac myocytes (CMs) using an in vivo rat model of Dox/Trz-related cardiotoxicity., Methods and Results: Adult rats received 6 doses of either Dox or Trz, or the two agents sequentially. Dox-mediated left ventricular (LV) dysfunction was aggravated by Trz administration. Dox treatment, but not Trz, induced T-tubule disarray. Moreover, Dox, but not Trz monotherapy, induced prolonged action potential duration (APD), increased incidence of delayed afterdepolarizations (DADs) and beat-to-beat variability of repolarization (BVR), and slower Ca 2+ transient decay. Although APD, DADs, BVR and Ca 2+ transient decay recovered over time after the cessation of Dox treatment, subsequent Trz administration exacerbated these abnormalities. Trz, but not Dox, reduced Ca 2+ transient amplitude and SR Ca 2+ content, although only Dox treatment was associated with SERCA downregulation. Finally, Dox treatment increased Ca 2+ spark frequency, resting Ca 2+ waves, sarcoplasmic reticulum (SR) Ca 2+ leak, and long-lasting Ca 2+ release events (so-called Ca 2+ "embers"), partially reproduced by Trz treatment., Conclusion: These results suggest that in vivo Dox but not Trz administration causes T-tubule disarray and pronounced changes in electrical activity of CMs. While adaptive changes may account for normal AP shape and reduced DADs late after Dox administration, subsequent Trz administration interferes with such adaptive changes. Intracellular Ca 2+ handling was differently affected by Dox and Trz treatment, leading to SR instability in both cases. These findings illustrate the specific roles of Dox and Trz, and their interactions in cardiotoxicity and arrhythmogenicity., Competing Interests: MF was employed by company Windtree Therapeutics Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Altomare, Lodrini, Milano, Biemmi, Lazzarini, Bolis, Pernigoni, Torre, Arici, Ferrandi, Barile, Rocchetti and Vassalli.)

Objective: To estimate the incidence and recurrence of breast cancer (BC) in patients with vulvovaginal atrophy (VVA) treated with ospemifene and matched untreated VVA patients using real-world data., Study Design: Retrospective matched cohort study., Main Outcome Measures: VVA patients were identified from the 2011-2018 US MarketScan® insurance claims database. For incidence, ospemifene-treated VVA patients without evidence of BC prior to index treatment were matched to two untreated VVA controls similarly without history of BC on age, index VVA year, geographic region, Charlson Comorbidity categories, and follow-up time. BC after the index treatment was identified by BC diagnosis codes, mastectomy, chemotherapy, or radiation procedure. Incidence rate, rate ratio (RR) and their 95 % confidence intervals (CI) were calculated. The process was repeated to estimate BC recurrence in patients with a history of BC in 1:1, 1:2 and 1:3 matches., Results: 1728 ospemifene users and 3456 untreated patients met the inclusion and matching criteria for assessing incidence. The average number of days for which ospemifene was supplied was 314 (standard deviation [SD] = 340). Average follow-up time from index treatment was 937 days (SD = 392) for treated patients and 915 days (SD = 396) for controls. BC incidence rates per 1000 person-years was 2.03 (95 % CI: 1.06-3.91) for treated patients and 3.53 (95 % CI: 2.49-4.99) for controls (RR = 0.58, 95 % CI: 0.28-1.21). No difference in recurrence was observed between ospemifene-treated and matched untreated patients. Ten (32.3 %) treated vs. 25 (40.3 %) controls in the 1:2 matched analysis had a recurrence., Conclusion: No differences were observed in the BC incidence and recurrence rates in ospemifene users compared with matched controls., (Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.)

A 61-year-old female underwent a right radical nephrectomy and a left nephron sparing surgery in 2014 due to renal cell carcinoma. A renal cell carcinoma local recurrence, 28 mm in size, centrally located in the left kidney was treated using cryoablation in 2016. In November 2018, computed tomography (CT) scan showed three nodules (maximum size 15 × 11 mm) in the left kidney, and CT-guided needle biopsy was performed. For multifocal recurrence and the anatomical site of these three nodules, a simultaneous reversible electrochemotherapy treatment was performed in April 2019. At 6-month CT control, no evidence of residual disease was found. Electrochemotherapy could be used to treat locoregional renal cell carcinoma recurrence when other ablative techniques are not suitable. LEVEL OF EVIDENCE: Level 4, Case Report.

Background: Most cases of breast lesions of uncertain malignant potential (B3) undergo surgical intervention. We aimed to analyze the outcome of B3 lesion subtypes in a large series of screen-detected cases., Methods: We screened 2,986 core needle biopsies to classify B3 lesions. Positive predictive values (PPVs) for malignancy were calculated for a comprehensive risk characterization according to clinicopathologic and morphologic variables., Results: B3 lesions comprised 35% atypical ductal hyperplasia (PPV = 20%), 16.7% flat epithelial atypia (PPV = 12%), 22.7% lobular neoplasia (PPV = 16.2%), 9% papillary lesion (PPV = 18.5%), 8.6% phyllodes tumor (PPV = 3.8%), and 8% radial scars (PPV = 4.1%) based on histopathologic diagnosis. Upgrade rates were 15.9% for calcifications, 13.7% for mass lesions, and 16.7% for architectural deformities, with 8.3% of malignant lesions classified as ductal carcinoma in situ and 6.7% as invasive cancers (PPV = 15%)., Conclusion: B3 lesions entail a heterogeneous risk of malignancy, and careful radiologic-pathologic correlation is required for optimal treatment.

Background: Induced pluripotent stem cell (iPSC)-derived cardiomyocytes (iPSC-CMs) are a unique source of human cardiomyocytes for cardiac disease modeling. Incomplete functional maturation remains a major limitation, however. One of the determinants of iPSC-CM maturation is somatic cell origin. We therefore compared iPSC-CMs derived from different somatic cell sources., Methods: Cardiac-derived mesenchymal progenitor cells (CPCs), bone marrow-derived mesenchymal stem cells (BMCs), and human dermal fibroblasts (HDFs) from same patients were reprogrammed into iPSCs and differentiated into iPSC-CMs. Expression of cardiac-specific genes, caffeine-responsive cells, and electrophysiological properties of differentiated cells were analyzed. To assess the contribution of epigenetic memory toward differences in gene expression observed during cardiac differentiation, DNA methylation patterns were determined in the early mesodermal cardiac promoter NKX2-5 and KCNQ1, which encodes for the pore-forming α-subunit of the slow component of delayed-rectifier potassium current (I Ks )., Results: Cardiac genes (MYH6, TNNI3, KCNQ1, KCNE1) were upregulated in CPC-vs. BMC- and HDF-iPSC-CMs. At early differentiation stages, CPC-iPSC-CMs displayed higher numbers of caffeine-responsive cells than BMC- and HDF-iPSC-CMs. The hERG1 (KV11.1) blocker, E4031, followed by the IKs blocker, JNJ303, increased extracellular field potential duration in CPC-iPSC-CMs to a greater extent than in BMC- and HDF-iPSC-CMs. The promoter region of NKX2-5 was more highly methylated in BMCs and HDFs compared to CPCs, and to a lesser extent in BMC-iPSCs compared to CPC-iPSCs., Conclusions: These results suggest that human iPSCs from cardiac somatic cell sources may display enhanced capacity toward cardiac re-differentiation compared to non-cardiac cell sources, and that epigenetic mechanisms may play a role in this regard., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Ligninolytic enzymes from white-rot fungi, such as laccase (Lac) and Mn-peroxidase (MnP), are able to degrade aflatoxin B 1 (AFB1), the most harmful among the known mycotoxins. The high cost of purification of these enzymes has limited their implementation into practical technologies. Every year, tons of spent mushroom substrate (SMS) are produced as a by-product of edible mushroom cultivation, such as Pleurotus spp., and disposed at a cost for farmers. SMS may still bea source of ligninolytic enzymes useful for AFB1 degradation. The in vitro AFB1-degradative activity of an SMS crude extract (SMSE) was investigated. Results show that: (1) in SMSE, high Lac activity (4 U g -1 dry matter) and low MnP activity (0.4 U g -1 dry matter) were present; (2) after 1 d of incubation at 25 °C, the SMSE was able to degrade more than 50% of AFB1 , whereas after 3 and 7 d of incubation, the percentage of degradation reached the values of 75% and 90%, respectively; (3) with increasing pH values, the degradation percentage increased, reaching 90% after 3 d at pH 8. Based on these results, SMS proved to be a suitable source of AFB1 degrading enzymes and the use of SMSE to detoxify AFB1 contaminated commodities appears conceivable., Competing Interests: The authors declare no conflicts of interest.

Reprogramming of adult somatic cells into induced pluripotent stem cells (iPSCs) has revolutionized the complex scientific field of disease modelling and personalized therapy. Cardiac differentiation of human iPSCs into cardiomyocytes (hiPSC-CMs) has been used in a wide range of healthy and disease models by deriving CMs from different somatic cells. Unfortunately, hiPSC-CMs have to be improved because existing protocols are not completely able to obtain mature CMs recapitulating physiological properties of human adult cardiac cells. Therefore, improvements and advances able to standardize differentiation conditions are needed. Lately, evidences of an epigenetic memory retained by the somatic cells used for deriving hiPSC-CMs has led to evaluation of different somatic sources in order to obtain more mature hiPSC-derived CMs.

Objective: To prospectively evaluate the effects of ospemifene on the vulva and vagina in postmenopausal women using vulvar-vestibular photography and direct visual assessments., Methods: Postmenopausal women (aged 40-80 years) with moderate to severe vaginal dryness as their most bothersome symptom (MBS) were randomized to daily ospemifene 60 mg or placebo in this 12-week, multicenter, double-blind, phase 3 study. Vulvar-vestibular photographic images were captured at baseline and week 12 and were independently assessed with the Vulvar Imaging Assessment Scale (VIAS). Changes from baseline in Vaginal and Vulvar Health Indices (VHI and VuHI) with ospemifene versus placebo were analyzed at weeks 4, 8, and 12. Correlations between VIAS, VHI, and VuHI, with vaginal dryness severity and the Female Sexual Function Index (FSFI) scores were also assessed., Results: In all, 631 eligible participants were randomized (ospemifene 316, placebo 315) and included in the intention-to-treat population. Compared with placebo, ospemifene significantly improved total scores for VIAS (P = 0.0154), VHI (P < 0.0001), and VuHI (P < 0.0001) from baseline to week 12; significant VHI (P < 0.0001) and VuHI (P = 0.002) improvements were observed at week 4. Most VHI and VuHI individual items were significantly better with ospemifene versus placebo at week 12 (P < 0.05). Most correlations between the vulvovaginal assessment total scores versus vaginal dryness severity and FSFI scores were significant (P < 0.05)., Conclusion: Improvements observed in vulvovaginal health with ospemifene assessed by prospective vulvar-vestibular photography and other direct visual assessments support its efficacy in addition to the treatment of moderate to severe vaginal dryness due to menopause and the use of photographic and direct visual evaluations in future clinical trials., Video Summary: Supplemental Digital Content 1, http://links.lww.com/MENO/A415.

Biochar (BC) and hydrochar (HC) are carbonaceous products obtained through, respectively, pyrolysis and hydrothermal carbonization processes of biomass. Both materials are multi-functional soil amendments. Ligninolytic fungi are primary decomposers of recalcitrant lignocellulosic material in nature through their extensive hyphal network and enzymes. In this work, two BC samples from red spruce pellets (BC SP ) and grapevine pruning residues (BC GV ) and two HC samples from urban pruning residues (HC UP ) and the organic fraction of solid urban wastes (HC SU ) were tested at concentrations of 0.4% and 2% (w/v) on the growth and enzyme activity of Trametes versicolor , Pleurotus ostreatus and Pleurotus eryngii . In all treatments with the lower concentration, BC and HC significantly stimulated fungal growth (up to about 90% increase for HC SU on T. versicolor ), whereas at the higher dose some inhibition was observed on T. versicolor by BC SP and P. ostreatus by BC SP , BC GV and HC UP . The two materials, especially HC, at both doses noticeably increased the activity of laccase from T. versicolor and P. eryngii , up to 21 and 13 times, respectively, for HC UP compared to controls. The activity of manganese peroxidase from P. ostreatus was also greatly stimulated by BC and HC, especially when added at the higher concentration. The overall results obtained in this study suggest potential benefits for ligninolytic fungi from the presence of these materials in soil at adequate dose of application.