A consistent finding of many studies describing the spectrum of mutant phenylalanine hydroxylase (PAH) alleles underlying hyperphenylalaninemia is the impossibility of achieving a 100% mutation ascertainment rate using conventional gene-scanning methods. These methods include denaturing gradient gel electrophoresis (DGGE), denaturing high performance liquid chromatography (DHPLC), and direct sequencing. In recent years, it has been shown that a significant proportion of undetermined alleles consist of large deletions overlapping one or more exons. These deletions have been difficult to detect in compound heterozygotes using gene-scanning methods due to a masking effect of the non-deleted allele. To date, no systematic search has been carried out for such exon deletions in Italian patients with phenylketonuria or mild hyperphenylalaninemia. We used multiplex ligation- dependent probe amplification (MLPA), comparative multiplex dosage analysis (CMDA), and real-time PCR to search for both large deletions and duplications of the phenylalanine hydroxylase gene in Italian hyperphenylalaninemia patients. Four deletions removing different phenylalanine hydroxylase (PAH) gene exons were identified in 12 patients. Two of these deletions involving exons 4-5-6-7-8 (systematic name c.353-?_912 + ?del) and exon 6 (systematic name c.510-?_706 + ?del) have not been reported previously. In this study, we show that exon deletion of the PAH gene accounts for 1.7% of all mutant PAH alleles in Italian hyperphenylalaninemics.
DNA Mutational Analysis/*methods ; Disease Progression ; Exons/genetics ; Gene Frequency ; Humans ; Italy ; Phenylalanine Hydroxylase/*genetics/metabolism ; Phenylketonurias/epidemiology/*genetics/physiopathology ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Deletion/genetics

OBJECTIVES: In this article, we aim to share our experience in the hospital reorganization made to conduct the SARS-CoV-2 vaccination campaign, based on the principles of flexibility and adaptability. STUDY DESIGN: A descriptive study. METHODS: The data concerning the organization of the vaccination campaign were taken from the operative protocol developed by the hospital dedicated task force, composed by experts in hygiene, public health, occupational medicine, pharmacists, nurses, hospital quality, and disaster managers. Data about the numbers of vaccine administered daily were collected by the Innovation and Development Operative Unit database. RESULTS: Vaccinations against COVID-19 started across the EU on the 27th of December 2020. The first phase of the vaccination campaign carried out in our hospital was directed to healthcare workers immunization including medical residents, social care operators, administrative staff and technicians, students of medicine, and health professions trainees. The second phase was enlarged to the coverage of extremely fragile subjects. Thanks to the massive employment of healthcare workers and the establishment of dynamic pathways, it was possible to achieve short turnaround times and a large number of doses administered daily, with peaks of 870 vaccines per day. From the 27th of December up to the 14th of March a total of 26,341 doses of Pfizer have been administered. 13,584 were first doses and 12,757 were second doses. From the 4th to the 14th of March, 296 first doses of Moderna were dispensed. It was necessary to implement adequate spaces and areas adopting anti-contagion safety measures: waiting area for subjects to be vaccinated, working rooms for the dilution of the vaccine and the storage of the material, vaccination rooms, post-vaccination observation areas, room for observation, and treatment of any adverse reactions, with an emergency cart available in each working area. CONCLUSIONS The teaching hospital of Pisa faced the beginning of the immunization campaign readjusting its spaces, planning an adequate hospital vaccination area and providing an organization plan to ensure the achievement of the targets of the campaign. This represented a challenge due to limited vaccine doses supplied and the multisectoral teams of professionals to coordinate in the shortest time and the safest way possible. The organizational model adopted proved to be adequate and therefore exploited also for the second phase aimed to extremely fragile subjects.
2019-nCoV Vaccine mRNA-1273 ; BNT162 Vaccine ; COVID-19/prevention & control* ; COVID-19 Vaccines/administration & dosage* ; Hospitals, Teaching/organization & administration* ; Humans ; Immunization Programs/organization & administration* ; Italy/epidemiology* ; SARS-CoV-2/immunology*

OBJECTIVETo investigate the prevalence of methicillin-resistant staphylococci (MRS) which is a potencial risk factor of transmission between animals and humans in different types of horses (harness racing-horses, breeding mares and riding-horses) and to compare the antimicrobial resistance of the isolates.

METHODSA total of 191 healthy horses, housed at different locations of the Campania Region (Italy), were included in the study. Nasal swab samples were collected from each nostril of the horses. The mecA gene was detected by a nested PCR technique. Antibiotic susceptibility was tested for each isolate.

RESULTSMRS was isolated from nasal samples of 68/191 (35.6%; 95% CI: 28.9%-42.9%) healthy horses. All isolates were coagulase-negative with the exception of two coagulase-positive MRS strains, identified as Staphylococcus aureus and Staphylococcus pseudintermedius, 2/83 (2.4%; 95% CI: 0.4%-9.2%). Interestingly, both coagulase-positive MRS isolates were from harness racing-horses. These horses also presented a significantly higher positivity for MRS (53.3%; 95% CI: 40.1%-66.1%) than the breeding mares and riding-horses groups. Antibiotic susceptibility testing showed difference between isolates due to different origins except for an almost common high resistance to aminopenicillins, such as ampicillin and amoxicillin.

CONCLUSIONSIt can be concluded that harness racing-horses may act as a significant reservoir of MRS as compared to breeding mares and riding-horses.

Animals ; Anti-Bacterial Agents ; pharmacology ; Coagulase ; Drug Resistance, Multiple, Bacterial ; Female ; Horse Diseases ; epidemiology ; microbiology ; Horses ; Italy ; epidemiology ; Male ; Methicillin-Resistant Staphylococcus aureus ; drug effects ; isolation & purification ; Microbial Sensitivity Tests ; veterinary ; Polymerase Chain Reaction ; veterinary ; Prevalence ; Staphylococcal Infections ; epidemiology ; microbiology ; veterinary