Antimicrobial resistance changing trends of Klebsiella pneumoniae isolated over the last 5 years.

The aim of this study was to describe the prevalence and epidemiology distribution of K. pneumoniae isolated at University Hospital of Campania "Luigi Vanvitelli," including the susceptibility evolution profile. Data on resistant phenotype strains, such as extended-spectrum-ß-lactamase (ESBL) producers and carbapenem-resistant K. pneumoniae (CRE) isolates, were also reported. K. pneumoniae strains were collected at the Complex Operative Unit (UOC) of Virology and Microbiology from different colonization and infection sites from January 2016 to December 2020. The highest rates of isolation were in urinary samples and in respiratory and wound swabs. Antibiotics susceptibility patterns showed more than 50% of the isolates resistant to cephalosporins, fluoroquinolones and penicillin. On the other hand, from 20% to 40% of K. pneumoniae strains were resistant to carbapenems and aminoglycosides. Based on our analysis, fosfomycin, ceftazidime/avibactam and ceftolozane/tazobactam are still therapeutic alternatives. Data analysis on carbapenem class evolution in 2016-2020 showed a significant increase in resistance rates (p<0.05). Increased rates in CRE and ESBL producing K. pneumoniae since 2017 were reported. Providing information on clinical characteristics and epidemiology data on contemporary K. pneumoniae evolution could help mitigate the spread of these isolates in our hospital and avert the endemic levels that have been observed in Southern Italy and in other European countries.

Prevalence and Antibiotic Resistance Profile of Bacterial Pathogens in Aerobic Vaginitis: A Retrospective Study in Italy.

Aerobic vaginitis (AV) is a vaginal infectious condition, characterized by a high inflammatory response and/or signs of epithelial atrophy, a decrease in the amount of Lactobacillus spp. and an increase in enteric origin bacteria. AV, often misdiagnosed, is difficult to treat due to the emerging spread of multi-drug resistant bacterial strains. The present study aimed to define the prevalence of AV, to detect causative bacteria and their antimicrobial resistance pattern. Women 10-95 years old, admitted to San Giovanni di Dio e Ruggi d'Aragona Hospital, Salerno, Italy (in the years 2015-2019) are included in the study. Bacterial identification and antibiotic susceptibility tests were carried out by VITEK® 2. Among 2069 patients, 1176 tested positive for microbial growth. A higher incidence of infection was found in the 55-64 age group. Among the pathogenic strains, 50.4% were Gram-negative, and 49.6% were Gram-positive. Escherichia coli (E. coli) (32.5%) was the most representative strain, followed by Enterococcus faecalis (E. faecalis) (29.4%), Klebsiella pneumoniae (K. pneumoniae) (7.8%) and Enterococcus faecium (E. faecium) (7.7%). E. coli showed high sensitivity to carbapenems and amikacin. K. pneumoniae carbapenems resistance was fluctuating over time. Alarming resistance to vancomycin was not recorded for Enterococci. Both strains were sensitive to teicoplanin, linezolid and tigecycline. Proper diagnosis and an effective therapeutic approach are needed to improve AV management.

Epstein-Barr Virus Seroprevalence and Primary Infection at the University Hospital Luigi Vanvitelli of Naples from 2007 to 2017.

Epstein-Barr virus (EBV) is a common herpesvirus that may cause asymptomatic infection or various diseases, such as mononucleosis, lymphoproliferative disorders and several cancers. Our objective was to estimate the prevalence of EBV among patients hospitalized in "Luigi Vanvitelli" University Hospital in the last 10 years. Our results showed that EBV seroprevalence in our geographical area was 65%. Seroprevalence increased gradually with age with no significant difference between females (49.42%) and males (50.58%). The seropositivity for primary infection was higher in patients about 5 years old, while seropositivity for past infection was predominant in patients of about 35 years old. These results underline that children in our country are still exposed to EBV. The development and the deeper use of an EBV vaccine in the early years of life could represent the solution for this infection.

Human leukocyte antigen-DR mismatch is associated with increased in-hospital mortality after a heart transplant.

OBJECTIVES: Although previous studies have investigated the effect of human leukocyte antigen matching on long-term outcomes after heart transplants, its role in the prognosis after a heart transplant remains unclear, particularly with respect to short-term survival. MATERIALS AND METHODS: We evaluated the human leukocyte antigen mismatch on in-hospital mortality of 158 consecutive patients who had undergone a heart transplant between 2000 and 2008. Human leukocyte antigens-A, -B, and -DR were determined by means of serologic and molecular techniques. Univariate analysis and a multiple logistic regression models evaluated the effect of human leukocyte antigen variants on mortality, independent of clinical variables. RESULTS: In-hospital mortality was 11.4%. Higher prevalence of acute kidney injury (50.0% vs 12.9%), higher levels of troponins 48 hours after transplant (15.6 ± 12.0 ng/mL vs 9.7 ± 9.4 ng/mL), prolonged ischemia (188.2 ± 32.5 min vs 162.6 ± 40.7 min), higher frequency of reoperation (61.1% vs 17.9%), and higher human leukocyte antigen-DR mismatch (1.61 ± 0.5 vs 1.30 ± 0.6) were found in patients who died. By logistic regression analysis, humanleukocyte antigen-DR mismatch is associated with in-hospital mortality (OR=5.159, 95% CI=1.348-19.754), independent of the effect of covariates such as recipient age, mismatch sex, mismatch human leukocyte antigen-A, human leukocyte antigen-B, acute kidney injury, reoperation, ischemia duration, and levels of troponins. CONCLUSIONS: Human leukocyte antigen-DR mismatch is associated with in-hospital mortality in heart transplant.

Current concepts in histocompatibility during heart transplant.

Sensitized candidates for heart transplant usually end up on a long waiting list and have an increased risk of rejection, graft loss, and incidence of cardiac allograft vasculopathy. An increasing number of studies have demonstrated the negative effect of preformed and posttransplant antibodies on graft survival. Thus, in sensitized patients, the combination of new, appropriate, desensitization protocols, and monitoring of posttransplant development of donor-specific antibodies may improve short-term and long-term outcomes. Introduction of more-sensitive and more-specific techniques for antibody detection provides a valid tool for assessing the degree of pretransplant HLA histocompatibility, and, therefore, predicting the results of crossmatch in sensitized patients, which are difficult to transplant. Currently, there are no accurate and standard methods to determine the functional characteristics of antibodies detected by solid-phase assay and, therefore, to predict their clinical relevance. Therefore, the future of heart transplantation requires a better understanding of tissue typing techniques and the effect of anti-HLA antibodies on clinical outcome to prevent discrimination against sensitized patients at the time of organ allocation.

Adult stem cells and the clinical arena: are we able to widely use this therapy in patients with chronic limbs arteriopathy and ischemic ulcers without possibility of revascularization?

The following paper is an overview on stem cells therapy in patients with peripheral vascular diseases. Recent research shows the ability of stem cells to develop and strengthen the collateral network in ischemic legs. Here, we discuss this clinical and therapeutic approach. To date, research has been mainly focused on patients with ischemic ulcers without possibility of revascularization. Non-invasive stem cell therapy has been proposed as an alternative to the amputation of such patients, but when the ulcers appear it is sometime too late. In our point of view, the selection of patients is a very important issue and we believe that the best candidate for this treatment is the patient with intermittent claudication before the development of ulcers. This choice could allow the optimization of results by the type of treated patients and not only by the type of infused cells. Indeed, several variables still remain to be elucidated for stem cell therapy, including the type of cells to be used, the infusion route, and more importantly, the stage of patients to be treated.

Effects of intracellular acidosis on endothelial function: an overview.

The endothelium represents the largest functional organ in the human body playing an active role in vasoregulation, coagulation, inflammation, and microvascular permeability. Endothelium contributes to maintain vascular integrity, intravascular volume, and tissue oxygenation promoting inflammatory network response for local defense and repair. Acid-basis homeostasis is an important physiologic parameter that controls cell function, and changes in pH can influence vascular tone by regulating endothelium and vascular smooth muscle cells. This review presents a current perspective of the effects of intracellular acidosis on the function and the basic regulatory mechanisms of endothelial cells.

Repeated immune and non immune insults to the graft after heart transplantation.

The clinical transplantation outcome is related to both effects of immunological and non immunological factors degenerating into hyperacute, acute and chronic rejection. Modern immunosuppressive treatments have resolved most events linked to acute rejection while long-term survival still remains the major problem after heart transplantation. The goal of personalized immunosuppressive therapy is to prevent rejection without inducing toxic effects. The aim of future studies could be to clarify the pathogenesis of chronic rejection and develop new and less toxic therapeutic approaches to induce "tolerance" to the graft without major side effects.

Kidney and heart interactions during cardiorenal syndrome: a molecular and clinical pathogenic framework.

The heart and kidney are physiologically interconnected. Cardiorenal syndrome (CRS) is a pathological disorder where acute or chronic dysfunction in one organ may induce dysfunction in the other one. Although classical studies have proposed a role for hypertension, dyslipidemia and endothelial dysfunction, CRS should be considered as a complex molecular interplay of neurohumoral pathway activation including the sympathetic nervous system, the renin angiotensin aldosterone axis, the endothelin system and the arginine vasopressin system. This activation may induce vascular inflammation, oxidative stress, accelerated atherosclerosis, cardiac hypertrophy and both myocardial and intrarenal fibrosis with progression of CRS treatment. More recently, epigenetics has opened new pathogenic molecular routes for CRS. This will lead to a more rapid development of novel, safe and effective clinical therapies.

CXCR4/YY1 inhibition impairs VEGF network and angiogenesis during malignancy.

Tumor growth requires neoangiogenesis. VEGF is the most potent proangiogenic factor. Dysregulation of hypoxia-inducible factor (HIF) or cytokine stimuli such as those involving the chemokine receptor 4/stromal-derived cell factor 1 (CXCR4/SDF-1) axis are the major cause of ectopic overexpression of VEGF in tumors. Although the CXCR4/SDF-1 pathway is well characterized, the transcription factors executing the effector function of this signaling are poorly understood. The multifunctional Yin Yang 1 (YY1) protein is highly expressed in different types of cancers and may regulate some cancer-related genes. The network involving CXCR4/YY1 and neoangiogenesis could play a major role in cancer progression. In this study we have shown that YY1 forms an active complex with HIF-1alpha at VEGF gene promoters and increases VEGF transcription and expression observed by RT-PCR, ELISA, and Western blot using two different antibodies against VEGFB. Long-term treatment with T22 peptide (a CXCR4/SDF-1 inhibitor) and YY1 silencing can reduce in vivo systemic neoangiogenesis (P < 0.01 and P < 0.05 vs. control, respectively) during metastasis. Moreover, using an in vitro angiogenesis assay, we observed that YY1 silencing led to a 60% reduction in branches (P < 0.01) and tube length (P < 0.02) and a 75% reduction in tube area (P < 0.001) compared with control cells. A similar reduction was observed using T22 peptide. We demonstrated that T22 peptide determines YY1 cytoplasmic accumulation by reducing its phosphorylation via down-regulation of AKT, identifying a crosstalk mechanism involving CXCR4/YY1. Thus, YY1 may represent a crucial molecular target for antiangiogenic therapy during cancer progression.

Modification of the detrimental effect of TNF-α on human endothelial progenitor cells by fasudil and Y27632.

Exposure of human endothelial progenitor cells (EPCs) to tumor necrosis factor-α (TNF-α) reduced their number and biological activity. Yet, signal transduction events linked to TNF-α action are still poorly understood. To address this issue, we examined the possible effect of fasudil and Y27632, two inhibitors of Rho kinase pathway, which is involved in endothelial dysfunction, atherosclerosis, and in- flammation. Results demonstrated that incubation with fasudil starting from 50 µM but not Y27632 determined a dose-dependent improvement of EPC number during exposure to TNF-α (P < 0.05 vs. TNF-α alone). Analysis of the signal transduction pathway activated by TNF-α revealed that the increased expression of p-p38 was not significantly altered by fasudil. Instead, fasudil blocked the TNF-α induced phosphorylation of Erk1/2 (P < 0.05 vs. TNF-α) as well as the inhibitor of Erk1/2-specific phosphorylated form, i.e., PD98059 (P < 0.05 vs. TNF-α). These results were confirmed by analysis of these kinases by confocal microscopy. Finally, 2D-DIGE and MALDI-TOF/TOF analysis of EPCs treated with fasudil revealed increased expression levels of an actin-related protein and an adenylyl cyclase associated protein and decreased expression levels of proteins related to radical scavenger and nucleotide metabolism. These findings suggest that fasudil positively affects EPC number and that other major signals might take part to this complex pathway.