New or interesting regional bryophyte records for Italian bryoflora

Based on recent bryophyte collections five records are added to the bryophyte flora of some Italian regions. The hepatic Riccia beyrichiana is new to Campania, Archidium alternifolium, Campylopus introflexus and Hedwigia stellata are new mosses to Lazio and Hylocomium splendens to Basilicata. In addition, the presence of Campylopus atrovirens and Pleuridium acuminatum in Lazio is confirmed after more than half a centur

Clinical considerations, management and treatment of fever of unknown origin caused by urachal cyst: a case report

INTRODUCTION: Urachal cysts are rare congenital anomalies that often prompt referral to the paediatric general surgeon because of their associated complications such as infection, abdominal pain and the young age at presentation. In this report we describe a rare case of fever of unknown origin caused by an urachal cyst which was successfully treated with incision and drainage only. Since the first description of urachal anomalies by Cabriolus in 1550, few cases have been reported and, until now, only one case of infected urachal cyst presenting as fever of unknown origin has been described in the literature. Moreover, the spontaneous resolution of an urachal cyst without excision is extremely rare. CASE PRESENTATION: We report our experience in the management and treatment of an infected urachal cyst that occurred in a 12-year-old Caucasian girl who presented to our Department of Paediatric Surgery with a 30-day history of evening fever. The urachal cyst was treated only with incision and drainage through a minimally invasive laparoscopic approach. CONCLUSIONS: The incision and drainage of an infected urachal cyst is a simple and safe procedure. It assures a complete recovery and avoids potential surgical complications related to the total excision of the urachal cyst. This report may provide important clues regarding the management of this rare anomaly and we emphasise the importance for paediatricians, who should consider the possibility that a fever of unknown origin can be caused by an urachal cyst, and for surgeons and urologists, because it suggests that conservative treatment of this rare anomaly should be considered when possible

Biofilm growth on orthopedic implantable materials: static or dynamic condition what is the most appropriate methodological tools to study device-related infections?

Aim Study of biofilm growth under static and dynamic conditions to evaluate the most suitable orthopedic materials on the prevention of device-related infections. Method Biofilms of Staphylococcus epidermidis (ATCC 35984) icaA and icaD genes positive and Pseudomonas aeruginosa (DSM 939) were generated under static and dynamic conditions, adding the bacterial inocula on titanium, carbon, polycarbonate and carbon-peek coupons housed in flat bottom test tubes or in the CDC Biofilm Reactor (CBR) system respectively. Biofilm growth was evaluated by MTT assay after 48 hours. Results Results of dynamic model showed a better capacity of S.epidermidis to grow with a rotation between 120-60 rpm on each tested materials (Mann-Whitney test, p-value < 0,05) than P.aeruginosa. Titanium was thematerial on which the bacterial strains adhered less, whereas carbon and polycarbonate allowed greatest adherence of P.aeruginosa (Mann-Whitney test, p-value < 0,05). Results of static model showed that both species grew on each materials without distinction (Kruskal-Wallis test, p-value 0,95). S.epidermidis growth was better also under static condition. Conclusions the static model was not able to evaluate the different adhesion capacity of the strains to the materials, confirming the dynamic model is the most suitable tool for the study of orthopedic materials on the prevention of device-related infections. This research was funded by the University of Pisa, PRA 2017_18 Projec

Collateral vascular network in a case of cardiac lymphoma: anatomo-clinical considerations

Primary cardiac tumours have an incidence of 0.2%1. Lymphomas represent 1.3% of all malignant primary cardiac tumours2 and are highly aggressive neoplasms. We describe a case of a rare primary cardiac tumor and the establishment of a collateral circulation. A 62-year-old white man referred to our clinical center with sinus tachycardia, congestion in the face and neck, pathologic jugular turgor, and hypovolemia in the superior caval system. Laboratory investigations revealed no abnormalities, except for a slight anemia and a moderate increase in carcinoembryonic antigen and in the lactate dehydrogenase level. The Multislice Computed Tomography (MSCT) revealed a right atrial infiltrative mass and superior vena cava (SVC) syndrome, due to obstruction of SVC drainage. In one week, after admission and without any therapy, congestion and turgor progressively disappeared because venous collaterals were recruited and dilated, offering alternative pathways through the parietal and mediastinal veins, from the superior (brachiocephalic system) to the inferior caval system (countercurrent or retrograde flow). On the other hand, from the inferior vena cava (IVC), the venous blood followed the normal route to the right atrium. In other words, three compensatory venous circles, anterior (ventral), posterior (dorsal), and confluent posterior-anterior, were constituted. A transjugular endomyocardial biopsy revealed that the mass was a large B-cell non-Hodgkin lymphoma, and positron emission tomography was positive for a primitive location, without any evidence of extracardiac involvement. The patient underwent 6 cycles of R-CHOP (rituximab, cyclophosphamide, doxorubicin, vincristine, prednisone) chemotherapy. After 8 months, CT venous phase venography with 64-detector CT angiography revealed that the superior caval-right atrium flow was restored, along with depletion of the anastomotic circles that had secured the return of retrograde flow through the IVC. After treatment, the patient experienced complete remission, and no recurrence has been observed to date. MSCT has proven to be very useful in the early detection of a cardiac mass and it allows for a noninvasive evaluation of the vascular network. In our case, no other older diagnostic tool would have been able to detect the reversal of venous blood flow from the cephalic district to the heart using the IVC. The images obtained by MSCT confirm that the vascular system responds to the functional needs of organs

Anti-proliferative effects of Cetuximab and Trastuzumab in colorectal cancer cell lines

Colon cancer is one of the most common human malignancies and a leading cause of death worldwide. In Europe around 250.000 new colon cases are diagnosed each year, accounting for around 9% of all the malignancies (Labianca R et al., 2010; Berrino F et al., 2007). Dysregulation of the signalling pathways induced through EGF receptors (ErbB/ HER receptors) by their over-expression or constitutive activation can promote tumor processes (Lurje G et al., 2009), including colorectal cancer. Therefore, the ErbB/HER receptor family with their most prominent members EGFR and HER-2 represents validated targets for anti-cancer therapy. Cetuximab and trastuzumab are two monoclonal antibodies approved for treating, respectively, metastatic colorectal and breast cancer. Because the monotherapy with cetuximab in metastatic colorectal is often insufficient (Cunningham D et al., 2004), it is useful to develop complementary therapeutic strategies to enhance antibody efficacy. A possible approach is co-administration of inhibitors, targeting multiple members of the EGF receptor family. In this study we examined the effect of cetuximab and trastuzumab in combination using two human colon cancer cell lines as a model. We observed that the two drugs had a cytostatic effect and inhibited the proliferation of both the cell lines in a time- and concentration-dependent manner. However, the combination had lower efficacy on one cell line than the other, with growth inhibition of 31% in the former and 49% in the latter. This result was associated to specific changes in cell cycle distribution, while no apoptosis was observed. Chromosome copy number heterogeneity and aneuploidy in tumoral cell lines have been reported (Pellestor F et al., 1999). Our data deriving from the cell cycle analysis confirmed the aneuploidy and polyploidy in our cellular models and are useful to explain cellular response to the combination. We used fluorescent in situ hybridisation analysis to evaluate EGFR and HER-2 gene amplification status. Both the tumour cell lines resulted in an abnormal copy number for the two genes resulting from aneuploidy (polisomy of chromosome 7 and 17) which is not responsible for the difference in sensitivity to cetuximab and trastuzumab between the two cell lines. In order to understand and to improve the pharmacological efficacy of cetuximab and trastuzumab combination, it will be useful to elucidate the molecular mechanisms involved in their activity. This will allow to develop novel and interesting approaches to cancer therapy

Recent advances in molecular diagnostics of colorectal cancer by genomic arrays: proposal for a procedural shift in biological sampling and pathological report

Two forms of genetic instability have been described in colorectal cancer: chromosomal instability, characterized by structural and numerical chromosomal abnormalities and associated to aneuploidy; and microsatellite instability, characterized by a deficiency in the mismatch repair system that leads to slippage in microsatellites and is associated to euploidy. Thirteen colorectal cancer sample DNAs were analyzed after colectomy. High-resolution genome-wide DNA copy number and Single Nucleotide Polimorphism genotyping analysis was performed by Affymetrix SNP 6.0 arrays that interrogates 906,600 single nucleotide polymorphisms and 945,826 copy number probes. We implemented this analysis as part of a routine procedure that includes the sampling of fresh tissue from the tumor mass without affecting the subsequent standard histopathological procedure. The novel molecular technology allows the determination of a genome-wide molecular karyotype using only 500 ng of high-quality tumor DNA; it distinguishes the two main types of genomic instability, discriminating between chromosomal instability positive and negative tumors. It also detects loss of heterozygosity (LOH) regions, called copy neutral-LOH. Tumor-associated copy neutral-LOH regions may play a pivotal role in oncogenesis when they determine duplications of either activating or loss of function gene mutation. We observed recurrent gains of chromosomes 2, 7, 8q, 9, 12, 13, 20 and losses of chromosomes 4, 5, 8p, 15, 17p, 18, 22, and Y, in agreement with previous cytogenetic studies. The use of such sampling procedure could stimulate the routine detection of point mutations in specific genes, thus avoiding subsequent sectioning of formalin-fixed and paraffin-embedded samples

Immunohistochemistry evidence for gastric ghrelin cells hypertrophy in obese patients

Ghrelin cells are endocrine cells of the gastrointestinal tract mainly present in the glands of the stomach. They produce the “hunger hormone” that promotes appetite and adiposity in animal and human models. In response to these anabolic effects, several elements have suggested the influence of ghrelin on the regulation of metabolic functions and the development of obesity-related disorders. However, its physiologic significance is quite complex and some of the effects of ghrelin are still debated in the literature. Recent evidence suggests that ghrelin influences glucose homeostasis through the modulation of insulin secretion and insulin receptor signalling. T2 diabetes is the most frequent metabolic disorder associated with obesity. To date, the most effective treatment of obesity is bariatric surgery and one of the most used techniques is sleeve gastrectomy. It consists in the resection of 25% of the stomach producing a neo anatomy in which this organ becomes a tube-like structure, lacking the ghrelinrich part. Forty-seven percent of T2 diabetic obese patients treated by sleeve gastrectomy rescue their diabetic condition just after surgery and before any weight reduction. This clinical evidence clearly connects this new morphology of the stomach with the positive metabolic improvement and suggests that ghrelin could play a key role in this striking effect. We studied the gastric mucosa of the fundus of sleeve gastrectomized T2 diabetic-obese patients (n.5) by immunohistochemistry with Anti- Ghrelin antibody to verify the ghrelin cell morphology, immunoreactivity and density. Anti- Chromogranin A antibody was used to detect the whole endocrine population of gastric glands. We used the normal part of the gastric fundus removed from patients (n.3) suffering for gastric cancer, as control. Ghrelin receptor has been shown on peripheral nerve fibers of the autonomous nervous system, and because a direct innervation of Langerhans islets by intestinal neurons have been shown in the past, we extended our immunohistochemistry analyses to neuronal antigens in order to study the nervous network of gastric mucosae in these surgical samples. Results show that ghrelin cells in T2 diabetic obese patients represented a consistent percentage (48.4% vs 38% in controls) of chromogranin A immunoreactive cell population. Furthermore, ghrelin cells were hypertrophic and intensely immunostained when compared with those found in gastric mucosae of controls. Neuronal specific enolase immunoreactivity was clearly enhanced in the gastric mucosae of obese patients. These data suggest that the stomach of T2 diabetic obese patients increase ghrelin content and that sleeve gastrectomy could have an important metabolic role by anatomical reduction of this hormone. Furthermore, our result suggest a possible paracrine effect on the peripheral neuronal population that could also be implicated in this complex and striking clinical effect of bariatric surgery

PIK3R1W624R Is an Actionable Mutation in High Grade Serous Ovarian Carcinoma.

Identifying cancer drivers and actionable mutations is critical for precision oncology. In epithelial ovarian cancer (EOC) the majority of mutations lack biological or clinical validation. We fully characterized 43 lines of Patient-Derived Xenografts (PDXs) and performed copy number analysis and whole exome sequencing of 12 lines derived from naïve, high grade EOCs. Pyrosequencing allowed quantifying mutations in the source tumours. Drug response was assayed on PDX Derived Tumour Cells (PDTCs) and in vivo on PDXs. We identified a PIK3R1W624R variant in PDXs from a high grade serous EOC. Allele frequencies of PIK3R1W624R in all the passaged PDXs and in samples of the source tumour suggested that it was truncal and thus possibly a driver mutation. After inconclusive results in silico analyses, PDTCs and PDXs allowed the showing actionability of PIK3R1W624R and addiction of PIK3R1W624R carrying cells to inhibitors of the PI3K/AKT/mTOR pathway. It is noteworthy that PIK3R1 encodes the p85α regulatory subunit of PI3K, that is very rarely mutated in EOC. The PIK3R1W624R mutation is located in the cSH2 domain of the p85α that has never been involved in oncogenesis. These data show that patient-derived models are irreplaceable in their role of unveiling unpredicted driver and actionable variants in advanced ovarian cancer

Pathogen-sugar interactions revealed by universal saturation transfer analysis

Many pathogens exploit host cell-surface glycans. However, precise analyses of glycan ligands binding with heavily modified pathogen proteins can be confounded by overlapping sugar signals and/or compounded with known experimental constraints. Universal saturation transfer analysis (uSTA) builds on existing nuclear magnetic resonance spectroscopy to provide an automated workflow for quantitating protein-ligand interactions. uSTA reveals that early-pandemic, B-origin-lineage severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike trimer binds sialoside sugars in an “end-on” manner. uSTA-guided modeling and a high-resolution cryo–electron microscopy structure implicate the spike N-terminal domain (NTD) and confirm end-on binding. This finding rationalizes the effect of NTD mutations that abolish sugar binding in SARS-CoV-2 variants of concern. Together with genetic variance analyses in early pandemic patient cohorts, this binding implicates a sialylated polylactosamine motif found on tetraantennary N-linked glycoproteins deep in the human lung as potentially relevant to virulence and/or zoonosis