Invasive behaviour of the marine Rhodophyta Sphaerococcus coronopifolius Stackhouse, in a hypereutrophic Mediterranean lagoon.

Between 2017 and 2022, sediment labile organic matter, physico-chemical and nutrient content of the water column, biomass and C, N, P, S content of Sphaerococcus coronopifolius, a Rhodophyta that produced vegetative blooms in an area of the Orbetello lagoon (Italy) not far from the effluents of two land-based fish-farms, were examined and compared with an area even further away from that source where the species was not found. In order to understand the reasons for an important mat development in only one specific area, microcosm experiments were also carried out. Results suggest the species developed in dense and extensive mats under high orthophosphate and nitrate nitrogen ion concentrations conditions, behaving as an opportunistic species.

Assessment of the causes of Hg bioaccumulation in the fish of a Mediterranean lagoon subject to environmental management interventions.

In a Mediterranean lagoon characterized by high levels of Hg in the sediment, Hg content in commercial fish species was determined, and, in order to establish Hg concentration in the water column, Diffusive Gradient in Thin films technique (DGT) was used. The results allowed to state (1) the disturbance of the surface sediment and its resuspension in the water column did not cause detectable increase in Hg releases, (2) the East basin acted more than the West one in contamination of fish species, (3) the small-sized sea-breams (<400 g) were more contaminated than larger sizes ones (>400 g). We concluded: (1) fish contamination may also depend on direct contact with releases of the contaminant and small, gregarious and less sedentary subjects are more likely to incur releases of Hg; (2) Hg releases can be greater in areas with a relatively low sedimentary organic matter load compared to areas subjected to high density macroalgal mat, regardless of the amount of Hg present in the sediments; (3) wind transport of plant masses in low energy storage areas, may constitute an increase factor of Hg in the sediment. A critical review of the results was made in comparison with the available literature reports and some hypotheses were proposed regarding the possible dynamics and availability of the contaminant.

Correction to: Biodegradation of naphthenic acids: identification of Rhodococcus opacus R7 genes as molecular markers for environmental monitoring and their application in slurry microcosms.

The published online version contains mistake in the author list for the author "M. Cappelletti" was incorrectly presented.

Ingenol mebutate-mediated reduction in p53-positive keratinocytes in skin cancerization field directly correlates with clinical response in patients with multiple actinic keratoses.

BACKGROUND: UV radiation represents the main risk factor for non-melanoma skin cancers. Chronic UV exposure induces 'p53 patches', i.e. clonal outgrowths of keratinocytes with high nuclear expression of mutated p53, which might progress to actinic keratosis (AK) and ultimately squamous cell carcinomas (SCCs). AIMS: Analysis of ingenol mebutate gel (150 and 500 mcg/g) effects in the reduction in 'p53 patches' inside skin cancerization field (CF) in patients with multiple AKs of face/scalp or trunk/extremities, in order to investigate whether the expected reduction in p53+ keratinocytes might have a direct role in the long-term AK reduction in treated areas. RESULTS: We enrolled n = 10 patients, treated with ingenol mebutate and evaluated at 2 and 6 months after treatment. We observed clinical responses in the majority of patients (n = 7), with AK reduction or complete clearance (n = 6 and n = 1, respectively). Notably, two patients did not respond to the treatment, and in one patient, after initial partial response, new lesion was recorded. In untreated skin CF samples (n = 3), we observed numerous p53+ keratinocytes, similar to those observed in invasive SCC samples (53.56 ± 8.79 and 74.34 ± 22.05, respectively; P = 0.2). After treatment, we observed a variable p53+ keratinocyte reduction in CF samples at 2 months (24.67 ± 31.19; P = 0.19). Importantly, the amount of p53+ keratinocytes strongly and directly correlated with AK number (R2  = 0.81). CONCLUSION: Untreated skin CF expresses high level of p53+ keratinocytes as invasive SCC. Ingenol mebutate is able to reduce p53+ keratinocytes with variable efficacy, this reduction degree directly correlating with clinical efficacy.

Ultramicrosurgery: A new approach to treat primary male genital lymphedema.

BACKGROUND: Lymphedema is a chronic pathology characterized by progressive swelling due to lymphatic dysfunction (1). Literature contains few studies that focus on male genital lymphedema. A variety of surgical techniques as part of the male genital lymphedema therapeutic strategy has been described. Supramicrosurgical lymphatico-venular anastomosis s-LVA, based on connecting lymphatic collectors to venules, has evidenced efficient outcomes thus far. However, the peculiarity of the genital area may lead to an innovative and even more accurate surgical technique as a treatment of male genital lymphedema: lymphatic pre-collectors located superficially over the fascial layer can be used to perform the ultramicrosurgical anastomosis. PURPOSE OF THE STUDY: In this paper, the authors report their experience of this new surgical concept based on anastomosing lymphatic precollectors to venules. METHODS: We performed a retrospective study from 2014 to 2016. Six male patients with primary genital lymphedema underwent ultramicrosurgical lymphatico-venular anastomosis in Siena University Hospital, Italy. RESULTS: Ultramicrosurgical lymphatico-venular anastomosis has evidenced positive outcomes in terms of prognosis, infectious complications, volume reduction, and quality of life. The average cellulitis rate dropped from 2.5 episodes a year to 0.5 episodes after surgical intervention. The mean satisfaction index passed from 1.33 before the intervention to 2.83. CONCLUSION: Ultramicrosurgical lymphatico-venular anastomosis represents a challenging physiological approach for male genital lymphedema with promising outcomes.

ALEX index enables detection of alien macroalgae invasions across habitats within a marine protected area.

A modified version of the ALien Biotic IndEX (ALEX) has been recently proposed to evaluate biological invasions in macroalgal assemblages. ALEX was applied in a Marine Protected Area where a recreational-fishing port is present testing the following hypotheses: ALEX increases with the distance from the port, it changes between the two directions off the port and it changes among three different habitats: Cystoseira beds, algal turf and dead matte of the seagrass Posidonia oceanica. A total of 78 native macroalgal taxa and 4 introduced species were found, the Chlorophyta Caulerpa cylindracea and the Rhodophyta Apoglossum gregarium, Acrothamnion preissii and Womersleyella setacea. All study sites were in high quality status highlighting that the assemblages investigated were at an early stage of NIS invasion. However, ALEX detected different values among conditions and habitats within the MPA, suggesting a local dynamics of NIS spread and different resistance to invasion of the investigated habitats.

T regulatory cells mediate immunosuppresion by adenosine in peripheral blood, sentinel lymph node and TILs from melanoma patients.

T regulatory cells (Tregs), involved in tumour tolerance, can generate Adenosine by CD39/CD73 surface enzymes, which identify four Tregs subsets: CD39+CD73- nTregs, CD39+CD73+ iTregs, CD39-CD73+ oTregs and CD39-CD73- xTregs. In melanoma patients, increased Tregs levels are detected in peripheral blood (PB), sentinel lymph node (SLN) and tumour infiltrating lymphocytes (TILs), but Adenosine role was not investigated yet. We examined total Tregs and Adenosine subsets in PB, SLN and TILs from melanoma patients (n = 32) and PB from healthy donors (HD; n = 10) by flow cytometry. Total Tregs significantly increased in stage III-IV patients PB, in SLN and TILs, as compared to HD/stage I-II patients. Tregs subsets analyses showed that: 1) PB nTregs significantly increased in SLN and decreased in TILs; 2) iTregs significantly increased in stage III-IV patients PB and further significantly increased in SLN and TILs; 3) PB oTregs and xTregs significantly decreased in SLN and TILs. Patients clinical features did not significantly influence total Tregs, except SLN excision order. Results confirmed Tregs role in melanoma progression and indicate Adenosine generation as a novel escape mechanism, being nTregs and iTregs increased in PB/SLN/TILs.

Could MRI visualize the invisible? An Italian single center study comparing magnetic resonance lymphography (MRL), super microsurgery and histology in the identification of lymphatic vessels.

OBJECTIVE: Aim of this study is to evaluate the possibility of limb magnetic resonance lymphography (MRL) to differentiate lymphatic vessels from pathological veins, collect a specimen of the identified lymphatic vessel during operations of super microsurgical lymphatic-venular anastomosis (s-LVA) and perform immunohistochemical stainings to confirm the nature of the collected vessels. PATIENTS AND METHODS: Twenty patients presenting lymphedema were enrolled in this study. Five patients reported lower limb lymphedema and 15 patients reported upper limb lymphedema. All patients had the indication for s-LVA and underwent preoperative MRL imaging of the affected limb. A total of 57 lymphatic vessels were identified by MRL and used to guide s-LVA: all these vessels have also been used to perform an intraoperative biopsy for immunohistochemical evaluation. RESULTS: A total of 53/57 vascular structures resulted compatible with lymphatic vessels at the immunohistochemical study performed with D2-40 antibody; 3/57 specimen showed the absence of the D2-40 antibody. A significant association was found between preoperative MRL and immunohistochemical marker D2-40 on collected specimen. CONCLUSIONS: Most of the articles in the international literature report the concomitant presence of both lymphatic and venous vessels at MRL. However, no one in literature describes the possibility to differentiate venous vessels from lymphatic vessels, and this is a crucial issue for the correct evaluation of the lymphatic system in patients with limb lymphedema undergoing a future surgical correction. In the present study, MRL allowed to identify active lymphatic vessels. MRL was predictive to determine preoperatory lymphatic vessels and to perform successful s-LVA in lymphedema patients. This is the first study to prove the nature of the vessels identified at the preoperative MRL with immunohistochemical stainings.

Our supramicrosurgical experience of lymphaticovenular anastomosis in lymphoedema patients to prevent cellulitis.

OBJECTIVE: Aim of this paper is to present our reduction of the frequency of cellulitis before and after supramicrosurgical lymphaticovenular anastomosis (s-LVA) in lymphoedema patients, and discuss the possibility to perform this technique outside Japan. PATIENTS AND METHODS: 37 patients affected by lymphoedema were enrolled. All patients received preoperative indocyanine green lymphography. Under local anaesthesia s-LVA was performed on all patients. All patients were followed for 1 year. Lymphoedema was staged using the lymphoedema staging classification recommended by the International Society of Lymphology. Cellulitis rate was recorded for all patients the year before and after the s-LVA. A t-test was used to evaluate differences in the frequency of cellulitis the year before surgery and the year following surgery. RESULTS: Cellulitis incidence decreased in all patients, with a mean 1.7 cases the year before s-LVA and 0.1 the year after s-LVA. A significant difference between preoperative and postoperative cellulitis rate was found (p = 0.0012). CONCLUSIONS: This study reports our s-LVA case series of lymphoedema patients. With the proper learning curve, s-LVA may be reproduced and lymphoedema patients may gain a better quality of life and a reduced cellulitis rate.

Cellulose nanocrystals are effective in inhibiting host cell bacterial adhesion.

The use of cellulose nanocrystals (CNCs) as a biomaterial able to inhibit host cell bacterial adhesion is described. Pre-incubation of E. coli ATCC 25922 with a suspension of 0.1% CNCs afforded a significant 2 log reduction of bacterial adhesion to the intestinal cell monolayer HT29. This effect could be attributed to the interaction between the CNCs and the bacterial cell surface as confirmed using fluorescence microscopy experiments.

Supramicrosurgical lymphatico-venular anastomosis (LVA) in treating lymphoedema: 36-months preliminary report.

OBJECTIVE: Lymphoedema of the extremities is a widespread pathological condition that mostly occurs as a complication of cancer resections, especially in women. Conventional therapy refers to conservative and physiotherapeutic approaches. Surgical strategies have been widely reported in the literature and are still challenging. Part of this literature focuses on the supra microsurgical lymphaticovenular anastomosis (LVA) technique. LVA is characterized by a high success rate, minimal invasivity and broad indications. Furthermore, this procedure can be performed under local anesthesia. PATIENTS AND METHODS: From October 2011 through October 2014, 69 patients affected by lymphedema underwent LVA surgery in Siena University Hospital, Italy. Preoperative and postoperative evaluations were taken. RESULTS: Totally, 366 anastomosis have been performed. The average rate was 5.3 anastomosis per patient. All patients registered a decrease in the size of the affected side. The average volume reduction was 50%. Patients also showed a reduction of lymphangitis episodes and reduction of compression garments class. Moreover, a satisfaction index was evaluated. The majority of patients (72.5%) was extremely satisfied of the surgery. CONCLUSIONS: LVA has demonstrated to be an effective surgical strategy to treat lymphoedema, especially in secondary cases in early stages. Although LVA is widely discussed in the literature, the majority of works relates to Japanese authors and few reports exist outside Japan. This paper represents the very first retrospective analysis of the adoption of LVA technique in Italy and one of the few outside Japan.

CD4+FOXP3+ T regulatory cells decrease and CD3+CD8+ T cells recruitment in TILs from melanoma metastases after electrochemotherapy.

Electrochemotherapy (ECT) represents an effective local treatment for skin unresectable melanoma metastases with high overall objective response rate. ECT is based on the combination of anti-neoplastic drugs administration and cancer cells electroporation. Whether ECT can also activate the immune system is a matter of debate, however a significant recruitment of dendritic cells in melanoma treated metastases has been described. Herein we investigated immediate and late effects of ECT treatment on T cell subsets in ECT-treated lesions by fluorescent immunohistochemistry. Biopsies from melanoma patients (n = 10) were taken before ECT (t0), at d1 and d14 from treatment. At t0, CD3+CD4+ T cells were the most represented T cells, well detected in the perilesional dermis, particularly at tumour margin, while CD3+CD8+ T cells were less represented. CD4+FOXP3+ T regulatory (Treg) cells were present in the perilesional dermis and within the lesion. ECT induced a significant decrease of CD4+FOXP3+ Treg cells percentage in the perilesional dermis, observed at d1 and at d14 (p < 0.001). CD3+CD8+ T cells frequency significantly increased at d14 from treatment in the perilesional dermis (p < 0.001). Furthermore calreticulin translocation to the plasma membrane, a hallmark of immunogenic cell death, was observed in metastatic cells after ECT. The data reported here confirm that ECT induces a local response, with a lymphoid infiltrate characterized by CD4+FOXP3+ Treg cells decrease and CD3+CD8+ T cells recruitment in the treated lesions. These results might contribute to design novel combinational therapeutic approaches with ECT and immunotherapy in order to generate a systemic long-lasting anti-melanoma immunity.

Effects of Costa Concordia shipwreck on epiphytic assemblages and biotic features of Posidonia oceanica canopy.

This research provides first information about Posidonia oceanica canopy in the area affected by Costa Concordia wreck. Percentage cover of algal and animal taxa on the leaves was estimated and biotic features of the meadow were measured in the period just after the shipwreck until its removal from the impacted site. Changes in epiphytic assemblages and some biotic features were detected in the Disturbed site compared with Control ones, highlighting effects due to the wreck presence and activities related to its removal. A temporary decrease of encrusting macroalgae and an increase of erected macroalgae and foraminifers, as well as a temporary increase of tip erosion of the canopy were detected in the Disturbed site. The obtained results were discussed and hypotheses about possible synergic effects occurred near the wreck were commented.

Microencapsulation of new probiotic formulations for gastrointestinal delivery: in vitro study to assess viability and biological properties.

The paper describes the preparation of new probiotic formulations based on chitosan-coated alginate microcapsules containing three different probiotic strains, Lactobacillus plantarum PBS067, Lactobacillus rhamnosus PBS070, and Bifidobacterium animalis subsp. lactis PBS075 taken individually and as a mixture of them. The effects of microencapsulation on the viability of the strains in conditions simulating the gastrointestinal tract and under industrial processes conditions were studied. In addition, an evaluation of their probiotic properties was also investigated by in vitro tests on the human intestinal cell line HT-29 to explore the effect of microencapsulation on health beneficial effect of the considered strains. Non-encapsulated cells were completely destroyed when exposed to simulated gastric juice and other stress conditions, while encapsulated cells exhibited a significantly higher resistance to artificial intestinal juice and heat and osmotic treatment. Moreover, in this study, the effect of the various microencapsulated probiotic strain formulations was compared with analogous formulations also containing the ß-glucan Pleuran. The microencapsulation effectively protected the selected bacteria, as single strain and as a mixture of the three strains in both the formulations with and without Pleuran, from simulating gastrointestinal tract and industrial process conditions in delivering the viable cells without any significant adverse effect on their functionalities. The comparative study of the immunomodulatory properties of each single strain and the mixture of the three strains revealed a synergistic effect of the probiotic mixture, but no appreciable difference between the two kinds of formulations could be detected, as the effect of Pleuran is covered by the higher potential of the probiotic strains.

Evaluation of the probiotic properties of new Lactobacillus and Bifidobacterium strains and their in vitro effect.

Probiotic ingestion is recommended as a preventive approach to maintain the balance of the intestinal microbiota and to enhance the human well-being. During the whole life of each individual, the gut microbiota composition could be altered by lifestyle, diet, antibiotic therapies and other stress conditions, which may lead to acute and chronic disorders. Hence, probiotics can be administered for the prevention or treatment of some disorders, including lactose malabsorption, acute diarrhoea, irritable bowel syndrome, necrotizing enterocolitis and mild forms of inflammatory bowel disease. The probiotic-mediated effect is an important issue that needs to be addressed in relation to strain-specific probiotic properties. In this work, the probiotic properties of new Lactobacillus and Bifidobacterium strains were screened, and their effects in vitro were evaluated. They were screened for probiotic properties by determining their tolerance to low pH and to bile salts, antibiotic sensitivity, antimicrobial activity and vitamin B8, B9 and B12 production, and by considering their ability to increase the antioxidant potential and to modulate the inflammatory status of systemic-miming cell lines in vitro. Three out of the examined strains presenting the most performant probiotic properties, as Lactobacillus plantarum PBS067, Lactobacillus rhamnosus PBS070 and Bifidobacterium animalis subsp. lactis PBSO75, were evaluated for their effects also on human intestinal HT-29 cell line. The obtained results support the possibility to move to another level of study, that is, the oral administration of these probiotical strains to patients with acute and chronic gut disorders, by in vivo experiments.

Lymphaticovenular anastomosis to prevent cellulitis associated with lymphoedema.

BACKGROUND: One of the complications of lymphoedema is recurrent cellulitis. The aim was to determine whether lymphaticovenous anastomosis (LVA) was effective at reducing cellulitis in patients with lymphoedema. METHODS: This was a retrospective review of patients with arm/leg lymphoedema who underwent LVA. The frequency of cellulitis was compared before and after surgery. The diagnostic criteria for cellulitis were a fever of 38·5°C or higher, and warmth/redness in the affected limb(s). RESULTS: A total of 95 patients were included. The mean number of episodes of cellulitis in the year preceding surgery was 1·46, compared with 0·18 in the year after surgery (P < 0·001). CONCLUSION: LVA reduced the rate of cellulitis in these patients with lymphoedema.

Genome Sequence of Rhodococcus opacus Strain R7, a Biodegrader of Mono- and Polycyclic Aromatic Hydrocarbons.

Rhodococcus opacus strain R7 (CIP107348) degrades several mono- and polycyclic aromatic hydrocarbons. Here, we present the high-quality draft genome sequence of strain R7, consisting of 10,118,052 bp, with a G+C content of 67.0%, 9,602 protein-coding genes, and 62 RNAs genes.

Genome Sequence of Rhodococcus sp. Strain BCP1, a Biodegrader of Alkanes and Chlorinated Compounds.

Rhodococcus sp. strain BCP1 cometabolizes chlorinated compounds and mineralizes a broad range of alkanes, as it is highly tolerant to them. The high-quality draft genome sequence of Rhodococcus sp. strain BCP1, consisting of 6,231,823 bp, with a G+C content of 70.4%, 5,902 protein-coding genes, and 58 RNA genes, is presented here.