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N. Figura [1] M. Valassina [2] F. Roviello [3] F. Pinto [3] D. Marrelli [3] R. Vernillo [3] P. Valensin [2] M. Valentini [2] C. Vindigni [4] [10/09] H. pylori [...]

Background: The infection by H. pylori (HP) strains with different structural types of the cagA gene, may determine different risk of gastric carcinoma (GC) development. As post-genomic approach to the study of H. priori is restricted to only a few strains, we carried out a comparative proteome analysis of HP with different cagA structural types, isolated from patients with different pathologies. Methods: The characteristics of the six strains studied and the patients' pathologies are reported in the table. Strains were grown simultaneously and protein extracts were submitted to two-dimensional polyacrylamide gel electrophoresis. Immobiline strips covering a pH range of 3-10 were used for isoelectrofocusing, while a polyacrytamide gel separating proteins in the Mr range of 200-8 kDa was used for the second dimension. The digitalized images of the silver stained gels were qualitatively and quantitatively analyzed by Melanie II 2d-PAGE software (Bio-Rad). Spot intensities were obtained in pixel units and normalized to the total absorbance of the gel. Increasing/decreasing indexes (fold change) were calculated as the ratio of spot intensity (relative volumes). Identification of protein spots was performed by N-terminal microsequencing, gel matching and immunoblotting. Results: We observed: a) a high variability in the expression of proteins; b) sharing of virulence determinants, such as UreB, Cag26, CagA, catalase, UreA, TagD, 26K antigen, HSPs, NapA, etc.; c) a maximal expression of CagA by GC strains, independently of the structural cagA type; d) a quantitatively different expression of CagA in strains from diffuse and intestinal GC; e) a decreased expression of proteins other than CagA in the cagA isogenic mutant. Discussion: the increased CagA expression by GC strains may be used for diagnostic/prognostic purposes. The role of proteins expressed by different cagA structural types is uncertain. [ABSTRACT FROM AUTHOR]

A protracted outbreak of New Delhi metallo-β-lactamase (NDM)-producing carbapenem-resistant Klebsiella pneumoniae started in Tuscany, Italy, in November 2018 and continued in 2020 and through 2021. To understand the regional emergence and transmission dynamics over time, we collected and sequenced the genomes of 117 extensively drug-resistant, NDM-producing K. pneumoniae isolates cultured over a 20-mo period from 76 patients at several healthcare facilities in southeast Tuscany. All isolates belonged to high-risk clone ST-147 and were typically nonsusceptible to all first-line antibiotics. Albeit sporadic, resistances to colistin, tigecycline, and fosfomycin were also observed as a result of repeated, independent mutations. Genomic analysis revealed that ST-147 isolates circulating in Tuscany were monophyletic and highly genetically related (including a network of 42 patients from the same hospital and sharing nearly identical isolates), and shared a recent ancestor with clinical isolates from the Middle East. While the bla NDM-1 gene was carried by an IncFIB-type plasmid, our investigations revealed that the ST-147 lineage from Italy also acquired a hybrid IncFIB/IncHIB-type plasmid carrying the 16S methyltransferase armA gene as well as key virulence biomarkers often found in hypervirulent isolates. This plasmid shared extensive homologies with mosaic plasmids circulating globally including from ST-11 and ST-307 convergent lineages. Phenotypically, the carriage of this hybrid plasmid resulted in increased siderophore production but did not confer virulence to the level of an archetypical, hypervirulent K. pneumoniae in a subcutaneous model of infection with immunocompetent CD1 mice. Our findings highlight the importance of performing genomic surveillance to identify emerging threats., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

Background: Telemedical wound care is one of the possible applications of teledermatology. The treatment of pediatric wounds needs frequent and periodic assessments of their local status and adjustment of dressings choice., Materials and Methods: We present our experience using telemedicine in the successful assessment and treatment of 19 pediatric patients at the OPBG, Rome . Photographs with a digital camera were taken and sent weekly by mail to a wound care specialist in Rome. This allowed the expert to diagnose and evaluate the wounds periodically., Results: In the shown cases, telemedicine allowed us to have an immediate evaluation and therapy adjustment. The quality of the images was good enough that the physician could regularly evaluate the status of the wound and immediately give his feedback to the parents. Of these 19, 13 patients (68%) experienced a wound resolution during the remote monitoring period. The satisfaction of the parents detected at 3, 6 and 12 months was found to be respectively 57%, 71%, 84%., Conclusion: Reducing transportation to the hospital to obtain a specialist advice, wound teleconsultation lowers health care costs and improve the quality of life for pediatric patients and their family, while still maintaining a high quality of pediatric wound care.

Background: Helicobacter pylori infection may increase the risk of both histotypes of gastric carcinoma (GC): intestinal (IGC) and diffuse (DGC). Polymorphism of the main H. pylori virulence factors, cagA and vacA, could have a different impact on the histological variety of GC., Methods: Two hundred and twenty-six H. pylori colonies were examined from 28 GC patients: 15 with IGC and 13 with DGC. DNA was extracted from bacteria and a PCR was performed using primers specific for cagA, the 3' cagA variable region and s and m regions of vacA., Results: There were 214 cagA+ strains and 55.61% of them were isolated from IGC cases; there were 12 cagA- colonies and they were all isolated from DGC (P<0.001). Most patients were infected by strains with more than one cagA structural type. No strains with a particular cagA type were found to be significantly more frequent in either histological variety of GC. A percentage of 43.90 of strains with vacA subtype s1/m1 were isolated from IGC, whereas 80.95% of vacA subtype s1/m2 strains were isolated from DGC cases (P<0.001)., Conclusion: Polymorphism of the main virulence genes of H. pylori may play different roles in the pathogenesis of IGC and DGC.

Thyroglossal duct cysts are one of the most common congenital abnormalities of the cervical region. Complications of these swellings are rare, and among these, appearance of a carcinoma has also been noted. We present a case of papillary carcinoma arising in a thyroglossal duct cyst in 20-year-old woman with a swelling of about 4 cm, located at the middle region of the neck over the hyoid bone. Our patient was treated using a modified Sistrunk operation, in which thyroidectomy proved crucial for the correct diagnosis and continuation of appropriate treatment. Our case confirms the difficulty in distinguishing a primitive thyroglossal duct carcinoma from a synchronous metastatic papillary carcinoma of the thyroid. This dilemma often remains unresolved.

Background: Infantile hemangioma is the most common vascular tumor in newborns, with an incidence from 12 to 23% among preterm infants with low weight at birth and a female to male ratio of 3:1. The head and neck is the most frequently affected area (60%), and the scalp is a typical site for such large lesions., Objective: We describe some clinical and medical aspects in comparison with the surgical approach to giant infantile hemangioma of the scalp., Methods: The indications to treatment are discussed. An outcome basis evaluation, by reviewing some clinical cases, is provided to help readers better understand when and how to undergo surgery safely., Conclusion: Early excision of huge infantile hemangioma of the scalp is the treatment of choice if feasible within 5 months of age.

Toscana virus (TOSV), a member of the Bunyaviridae family, is an important etiologic agent of neurologic infection transmissible to humans by bites of the Phlebotomus spp. In consideration of the variations in the antigenic properties of Bunyaviruses and their potential genetic variability, we analysed a large region (2500nt) of the Toscana virus M segment coding for the non-structural protein (NSm) and the G(N) and G(C) glycoproteins in several strains isolated from patients with meningitis from 1998 to 2004 in the region of Tuscany in Italy. The sequences were compared with the reference strain of Toscana virus isolated from phlebotomus (ISS Phl. 3) and revealed some changes in amino acids, particularly in the G(C) protein, that are probably involved in recognition and binding to the cell receptor. The analyses were aimed at identifying the amino acid changes commonly to all of the clinical isolates potentially related to TOSV virulence.

Toscana virus (TOSV) has recently been recognized as an emerging virus transmitted by phlebotomus vectors, responsible for acute neurological diseases in Mediterranean countries. In our study, we demonstrated that adult Balb/c mice were susceptible to TOSV when infected intracerebrally (i.c.) or subcutaneously (s.c.) with a neuroadapted strain of the virus. We have shown that by performing serial passages of a wild type human isolate of TOSV in mouse brains, selection occurs for a highly virulent variant which replicates efficiently in the central nervous system (CNS) of i.c.-injected mice, causing acute encephalitis and death. Immunohistochemical analysis and TUNEL assay of post-mortem organs showed that TOSV replication was highly restricted to neurons in which it induced apoptotic death; however, virus antigen-positivity was also observed in the spleen and lymph nodes. In s.c.-injected mice, virus was detectable in the spleen and lymph nodes, whereas only few meningeal cells and neurons were affected, allowing for the mouse survival the infection. The presence of TOSV in spleen and lymph node cells in both s.c.- and i.c.-treated mice suggests their possible involvement in the diffusion of the infection. This animal model may be helpful for the development of prophylactic measures against TOSV infections.

Purpose of Review: Sandfly fever viruses are still a significant health problem in many regions of the world, such as Africa, the Mediterranean basin, the Middle East, and Central Asia. This review provides an update on the advances in knowledge about epidemiological, clinical, and laboratory aspects of infections caused by Toscana, Sicilian and Naples viruses., Recent Findings: Diagnosis of Toscana virus infection has been facilitated by new molecular methods and by immunoenzymatic tests based on the recombinant nucleoprotein. Gene analysis has allowed identification of circulating Toscana variants possibly involved in the protean pathomorphism and extreme variability of the clinical picture. New attention has been addressed to the antigenic properties of the viral proteins (the nucleoprotein N and the surface glycoproteins G1 and G2), in order to understand their immunogenetic role. High genetic divergence within the serocomplexes belonging to each of the Sicilian and the Naples viruses has suggested that infection with one genotype may not completely immunize against infection with all other genotypes in a given serocomplex. These findings could serve as a basis for vaccine development and may account for reports of multiple episodes of sandfly fever in the same host. Recently, the performance of analysis models based on weather data and reported vector surveys has allowed the prediction of the risk of acquiring sandfly infection in the endemic geographic areas., Summary: Recent developments include a better knowledge of the epidemiological, clinical, and laboratory aspects of sandfly infection, while the search for effective drugs and vaccines is still in progress.

We describe an outbreak of familial infection of Chlamydia pneumoniae, an etiological agent for respiratory tract infections. In a family member detection of C. pneumoniae on a pharyngeal swab by polymerase chain reaction was positive until four months after the onset of symptoms, despite a course of antibiotics known to be effective against Chlamydia species

Toscana virus is the most important agent responsible for meningitis in central Italy. We report a serosurveillance study, using an immunoenzymatic assay, of 360 serum samples harvested from a high-risk population occupationally exposed to Toscana virus in two regions of Italy, Tuscany and Piedmont. The results indicates a seroprevalence of Toscana virus of 77.2% in the forestry workers, particularly in the Tuscany region. This fact is strictly correlated with the ecological niches specific for the survival of Toscana virus arthropod vector.

In recent years, 16S ribosomal DNA analyses has allowed the recognition of new chlamydia organisms, requiring the creation of new species, genera, and families within this unique, deep lineage of prokaryotes. The trachoma and psittaci groups chlamydiae are now recognized as separate genera, Chlamydia and Chlamydophila, respectively, and biovars of each group have been elevated to the species rank. Simkania and Parachlamydia have been associated with human respiratory infections, while Waddlia seems to be implicated in abortion in bovins. DNA amplification studies targeting the 16S rDNA have revealed a richer diversity within chlamydiae, identifying new lineages from both environmental and clinical samples. Further studies will be of interest to both examine the ecology and evaluate the clinical importance of these novel chlamydiae. Herein, we provide a summary of literature and our data about the novel chlamydial lineages.

Chlamydiales are important intracellular bacterial pathogens, causing a wide variety of diseases in vertebrates, including humans. Besides the well-known species in the family Chlamydiaceae, new chlamydial organisms have recently been discovered, forming three new families: Parachlamydiaceae, Simkaniaceae and Waddliaceae. Parachlamydia acanthamoebae and Simkania negevensis are currently investigated as emerging human respiratory pathogens. Additional chlamydial lineages have been discovered by 16S rDNA-based molecular studies, and their implication in human infections is poorly known. By using a pan-chlamydia 16S rDNA PCR, we have searched for the presence of chlamydiae in 228 clinical samples that all previously had been shown to be PCR-negative for Chlamydophila pneumoniae: 170 respiratory samples, 45 atheromatic plaques and 13 peripheral blood mononuclear cell samples. Nine respiratory samples tested positive. Sequence analysis has allowed us to assign four sequences to Chlamydophila psittaci, three sequences to Chlamydophila felis, and two sequences to two novel phylotypes belonging to the Parachlamydiaceae. These latter sequences showed similarity values of more than 93% with each other and with the P. acanthamoebae sequence, thus belonging to novel, unrecognized species. In conclusion, this report showed that a variety of non-C. pneumoniae chlamydial respiratory infection is present in humans, and that new parachlamydiae distinct from P. acanthamoebae may be detected in human clinical samples. Future studies will be of interest in order to estimate the diversity of these novel chlamydiae in both clinical and environmental samples, as well as their possible clinical implication in human and animal infections.

Acute meningitis is the most common neurologic disease that involves the central nervous system. The spectrum of infectious agents that cause neurologic infection is remarkably broad and numerous viruses are the most frequent cause of the aseptic meningitis syndrome. We applied a multiplex one-step method for the rapid detection of the genomic RNA of different neurotropic viruses: particles in the genus Enterovirus and Toscana virus, which are the most representative aetiologic agents in our country during the spring-summer period. We have evaluated the sensitivity and the specificity of the multiplex one-step test on positive controls and on RNA extracted from clinical samples harvested from 475 patients with meningitis hospitalized during the 1996-2001 period. The multiplex one-step RT-nPCR protocol allows for the detection of enterovirus and Toscana virus RNA in a single sample, by using, at the same time, a very small clinical sample volume. In our study we were able to diagnose 192 cases of meningitis by Toscana virus and 31 cases by enteroviruses out of 475 cases of meningitis utilizing the described one-step multiplex method.