Co-infection of Streptococcus pneumoniae in Respiratory Infections Caused by SARS-CoV-2

Abstract: Viral respiratory infections are often associated with bacterial co-infections that often lead to increased severity and mortality of the disease. During the recent pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), hospitalized patients reported developing secondary bacterial infections ranging from 0 to 40% of the cases. In the previous influenza pandemics, Streptococcus pneumoniae was the most isolated bacterial pathogen causing increased mortality in patients affected by viral pneumonia. Due to the difficulty to detect pneumococcal infection in SARSCoV-2 patients by a rapid clinical test, the real prevalence of S. pneumoniae might be underestimated, and only a few cases have been documented so far. It has been estimated that 90% of patients admitted to the Intensive Care Unit are empirically treated with antimicrobial. The application of more rapid and sensitive diagnostic methods could help with targeted antibiotic therapy. Additionally, pneumococcal vaccination of high-risk individuals could reduce bacterial pneumonia, hospital admissions, and comorbidities associated with serious illness

Evaluation of “Caterina assay”: An Alternative Tool to the Commercialized Kits Used for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Identification

Abstract: Here we describe the first molecular test developed in the early stage of the pandemic to diagnose the first cases of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in Sardinian patients in February–March 2020, when diagnostic certified methodology had not yet been adopted by clinical microbiology laboratories. The “Caterina assay” is a SYBR®Green real-time reverse-transcription polymerase chain reaction (rRT-PCR), designed to detect the nucleocapsid phosphoprotein (N) gene that exhibits high discriminative variation RNA sequence among bat and human coronaviruses. The molecular method was applied to detect SARS-CoV-2 in nasal swabs collected from 2110 suspected cases. The study article describes the first molecular test developed in the early stage of the declared pandemic to identify the coronavirus disease 2019 (COVID-19) in Sardinian patients in February–March 2020, when a diagnostic certified methodology had not yet been adopted by clinical microbiology laboratories. The assay presented high specificity and sensitivity (with a detection limit ≥50 viral genomes/µL). No false-positives were detected, as confirmed by the comparison with two certified commercial kits. Although other validated molecular methods are currently in use, the Caterina assay still represents a valid and low-cost detection procedure that could be applied in countries with limited economic resource

Molecular pathways triggered by COVID-19 in different organs: ACE2 receptor-expressing cells under attack? A review

OBJECTIVE: In human pathology, SARS-CoV-2 utilizes multiple molecular pathways to determine structural and biochemical changes within the different organs and cell types. The clinical picture of patients with COVID-19 is characterized by a very large spectrum. The reason for this variability has not been clarified yet, causing the inability to make a prognosis on the evolution of the disease. MATERIALS AND METHODS: PubMed search was performed focusing on the role of ACE 2 receptors in allowing the viral entry into cells, the role of ACE 2 downregulation in triggering the tissue pathology or in accelerating previous disease states, the role of increased levels of Angiotensin II in determining endothelial dysfunction and the enhanced vascular permeability, the role of the dysregulation of the renin angiotensin system in COVID-19 and the role of cytokine storm. RESULTS: The pathological changes induced by SARS-CoV-2 infection in the different organs, the correlations between the single cell types targeted by the virus in the different human organs and the clinical consequences, COVID-19 chronic pathologies in liver fibrosis, cardiac fibrosis and atrial arrhythmias, glomerulosclerosis and pulmonary fibrosis, due to the systemic fibroblast activation induced by angiotensin II are discussed. CONCLUSIONS: The main pathways involved showed different pathological changes in multiple tissues and the different clinical presentations. Even if ACE2 is the main receptor of SARS-CoV-2 and the main entry point into cells for the virus, ACE2 expression does not always explain the observed marked inter-individual variability in clinical presentation and outcome, evidencing the complexity of this disorder. The proper interpretation of the growing data available might allow to better classifying COVID-19 in human pathology

Periodontal microbiota of Sardinian children: comparing 200-year-old samples to present-day ones

Introduction: The microrganisms of the human oral cavity include more than 700 species or phenotypes of bacteria. Some “diseases of civilization” are strictly correlated to changes in the microbiome following the food revolution that occurred after WWII. For that reason, a precise recognition of the microbiome profile before and after this period should be useful to determine the health-compatible model of icrobiome. The aim of this study was to compare the microbiome profiles (number of total cells, and pathogen types) of dental samples obtained from two distinct groups of children, a 200-year-old retrieved one and a present one. Methods: Two different groups of samples have been studied. The first group was a set of 50 recent subgingival plaque samples obtained from children of age 2-8 years, 14 males and 36 females. They were enrolled by the Department of Dental Disease Prevention (University of Cagliari, in Sardinia, Italy) during standard dental care procedures. None reported periodontal disease and none had been under antibiotic therapy during the previous 6 months. The second group was an old retrieved group that included 24 teeth from 6 different 6- to 8-year-old crania fragments; they were obtained from a 200-year-old charnel-house located in Villaputzu, a city close to Cagliari. Representative periodontal bacteria have been identified by a previously published real-time PCR procedure (Sokransky et al., 1998) in which P. gingivalis and T. forsythia (red complex), A. Original article 2/5 www.jpnim.com Open Access Journal of Pediatric and Neonatal Individualized Medicine • vol. 6 • n. 1 • 2017 Orrù • Contu • Casula • Demontis • Blus • Szmukler-Moncler • Serreli • Maserati • Steri • Fanos • Coghe • Denotti actinomycetemcomitans (green complex) and F. nucleatum (orange complex) were detected. In addition, the title of each pathogen was expressed as a percentage of the total bacteria (biofilm) in the sample. Results and discussion: The profile of periodontal microbiomes, between recent/ancient samples showed a significant difference relative to Sokransky’s red complex bacteria (p < 0.05). In all analyzed periodontal strains, the pathogenic bacteria P. gingivalis and T. forsythia showed the highest title in the recent group. Conclusions: Our hypothesis is that the transfer of “commensal-pathogen” as an absolute number on the oral biofilm might be linked to the distinct alimentary habits of the two populations. Some diet rich in reducing agents, such as processed meat-based foods, might be able to increase the average number of pathogen anaerobic bacteria in the oral microbiota. The outcome would be an increase of the oral systemic diseases reported with these pathogens. Our data suggest that the ancient Sardinian population was able to control the pathogen oral anaerobic biofilm by some diet rich in antioxidant compounds. Further investigations are required to focus on the genetic profile and the health status of this ancient population but it appears that molecular microbiology might be considered as the “time machine” in oral biology

The perception of professionals and users of the quality of care and respect for human rights in four outpatient care facilities of an Italian hospital during the Covid-19 pandemic

Background: During the Covid-19 pandemic, delays in providing medical services, dissatisfaction, criticism toward health workers (HW) and the risk of burnout of HW in Italy have been documented. No studies have contrasted the point of view of HW and users on the quality of care and respect for human rights in health facilities. Objective: To compare the perception of users of their satisfaction with the care provided with the perception of HW of their satisfaction with work as well as the perception of the respect of HW “s and users” human rights. Methods: The “Well-Being at work and respect for human rights questionnaire” (WWRR) was administered on a sample of users (142) and HW (154) in four outpatient health care facilities of a hospital in Sardinia, Italy. Results: Users showed higher scores than HW on their satisfaction with the care received (p &lt; 0.0001), the perception of respect for their human rights (p &lt; 0.0001), and availability of resources for care (p &lt; 0.0001). The HW scores were higher than 50% of the maximum in all items, but a relatively low score was reported on the HW’s satisfaction of the resources and the respect for their rights. Conclusion: The satisfaction for care and respect for human rights in the outpatient health services was higher than expected. The relatively low score by the HWs in relation to the satisfaction with the resources and perception of respect for their human rights could be a wake-up call. The study does not involve emergency rooms, wards, or Covid units

Evaluation of humoral and cellular response to third dose of BNT162b2 mRNA COVID-19 vaccine in patients treated with B-cell depleting therapy

Objective: to investigate the responses to mRNA COVID-19 vaccines in a cohort of immunosuppressed patients affected by immune-mediated inflammatory diseases (IMID). Methods: we have measured humoral and cellular immunity using quantitative IgG anti-SARS-CoV-2 Spike antibody (anti-S-IgG), neutralization assays and specific interferon-gamma (IFN-g) release assay (IGRA) before and after the third dose of BNT162b2. The response of those on anti-CD20 (n = 18) was then compared with healthy controls (HC, n = 18) and IMID naïve to anti-CD20 drugs (n = 13). Results: a third BNT162b2 dose is highly immunogenic in IMID patients naïve to anti-CD20, as 100% of the subjects seroconverted compared to the 55% in anti-CD20. The rate of IGRA response was of 79% in anti-CD20, 50% in IMID naïve to anti-CD20, 100% in HC. Among those who have seroconverted, IMID patients had significantly reduced anti-S-IgG and neutralization titers compared to HC, whereas no significant difference was observed when comparing anti-CD20 and HC. Furthermore, 13% of anti-CD20 and 7.7% of IMID were simultaneously negative for both neutralizing antibodies and IGRA after three doses. Conclusion: these data draw attention to the immunogenicity of COVID-19 vaccination in treated IMID, taking specific groups into consideration for vaccination program

Evaluation of antibody response to BNT162b2 mRNA COVID-19 vaccine in patients affected by immune-mediated inflammatory diseases up to 5&nbsp;months after vaccination

SARS-CoV-2 vaccination with mRNA product BNT162b2 elicited high immunogenicity in healthy subjects in trials. This study aims to better understand the factors that influence the humoral immune response to vaccination against SARS-CoV-2 in patients with immune-mediated inflammatory diseases (IMIDs). We enrolled patients and healthy healthcare workers control group (HCW) that underwent mRNA BNT162b2 vaccination and measured the serum IgG anti-S-RBD response at booster dose (T1), one month after booster dose (T2) and up to 5&nbsp;months (T3). Demographic, disease-specific and vaccination data were recorded. Vaccination response of 551 participants naïve to SARS-CoV-2 infection were included in HCW and 102 in the IMID group, analyzing separately those on anti-CD20. At T2 all naïve HCW developed anti-S-RBD-IgG, while 94% of IMID responded (p &lt; 0.001). IMID patients had a significantly different level of IgG than HCW at both T1 (p = 0.031), T2 (p &lt; 0.001), while there was no significant difference at T3. There were no statistically significant differences according to the IMID type or to ongoing treatment with immunosuppressants, corticosteroids or biological drugs other than anti-CD20. The proportion and magnitude of response was significantly lower in IMID treated with anti-CD20 drugs. There was a correlation with age at T1 and at T2 but not at T3, stronger in patients than in HCW. Immune response close after BNT162b2 vaccination is reduced in patients with IMID, but there is no significant difference at 5&nbsp;months. The measured reduction is related to age and the disease itself rather than treatments, with the exception of anti-CD20 drugs

Natural killer-cell immunoglobulin-like receptors trigger differences in immune response to SARS-CoV-2 infection

Background: The diversity in the clinical course of COVID-19 has been related to differences in innate and adaptative immune response mechanisms. Natural killer (NK) lymphocytes are critical protagonists of human host defense against viral infections. It would seem that reduced circulating levels of these cells have an impact on COVID-19 progression and severity. Their activity is strongly regulated by killer-cell immuno-globulin-like receptors (KIRs) expressed on the NK cell surface. The present study's focus was to investigate the impact of KIRs and their HLA Class I ligands on SARS-CoV-2 infection. Methods: KIR gene frequencies, KIR haplotypes, KIR ligands and combinations of KIRs and their HLA Class I ligands were investigated in 396 Sardinian patients with SARS-CoV-2 infection. Comparisons were made between 2 groups of patients divided according to disease severity: 240 patients were symptomatic or paucisymptomatic (Group A), 156 hospitalized patients had severe disease (Group S). The immunogenetic characteristics of patients were also compared to a population group of 400 individuals from the same geographical areas. Results: Substantial differences were obtained for KIR genes, KIR haplotypes and KIR-HLA ligand combinations when comparing patients of Group S to those of Group A. Patients in Group S had a statistically significant higher frequency of the KIR A/A haplotype compared to patients in Group A [34.6% vs 23.8%, OR = 1.7 (95% CI 1.1-2.6); P = 0.02, Pc = 0.04]. Moreover, the KIR2DS2/HLA C1 combination was poorly represented in the group of patients with severe symptoms compared to those of the asymptomatic-paucisymptomatic group [33.3% vs 50.0%, OR = 0.5 (95% CI 0.3-0.8), P = 0.001, Pc = 0.002]. Multivariate analysis confirmed that, regardless of the sex and age of the patients, the latter genetic variable correlated with a less severe disease course [ORM = 0.4 (95% CI 0.3-0.7), PM = 0.0005, PMC = 0.005]. Conclusions: The KIR2DS2/HLA C1 functional unit resulted to have a strong protective effect against the adverse outcomes of COVID-19. Combined to other well known factors such as advanced age, male sex and concomitant autoimmune diseases, this marker could prove to be highly informative of the disease course and thus enable the timely intervention needed to reduce the mortality associated with the severe forms of SARS-CoV-2 infection. However, larger studies in other populations as well as experimental functional studies will be needed to confirm our findings and further pursue the effect of KIR receptors on NK cell immune-mediated response to SARS-Cov-2 infection

L'IMPIEGO DEI BIOMARCATORI SIERICI NELLA GESTIONE CLINICA DELL'ENDOMETRIOSI

Endometriosis is a frequent gynecological disorder that affects 3% to 15% of the general premenopausal female population. Recently, some studies have suggested that women suffering from endometriosis, especially if ovarian endometriosis (endometrioma), have a risk increased up to eight times compared to the general female population to develop ovarian cancer or borderline ovarian tumor. The HE4 marker was proposed in combination with the CA125 and the menopausal status of the woman to calculate the risk that a pelvic mass may be malignant. However, very high concentrations of CA125 are present in women with endometrioma and in women with benign deep infiltrating endometriosis. It was therefore decided to verify the efficacy of the HE4 marker alone in identifying cases of endometrioma with proven histological benignity. Using 70 pmol / L as the reference cutoff as per literature, only 10.3% (i.e. 3 patients out of 28) showed HE4 values higher than the cut-off. However, if the cut-off value had been raised to 80 pmol / L, 100% of the patients would have had lower values than this cut-off

CYTOMEGALOVIRUS INFECTION IN THE FIRST DAYS OF LIFE: PREVALENCE IN THE SOUTH SARDINIA FROM 2016 TO 2019

Congenital cytomegalovirus (CMV) infection poses a significant health risk to women of childbearing age and pregnant women and their unborn babies. Screening for CMV represents a crucial objective for Public Health, since this virus is one of the leading causes of childhood disability. CMV belongs to the Herpesviridae family and, once contracted the infection, it remains latent inside the organism for life, but it could be reactivated in case of weakening of the immune system. CMV infections are generally asymptomatic, but in immunocompromised patients, they can cause serious complications. CMV can be transmitted vertically from mother to child, as well as a primary, recurrent, or chronic infection. Maternal-fetal transmission of CMV can occur in the uterus (congenital infection), during labor or delivery (perinatal infection), or by lactation (postnatal infection) and it can cause miscarriage, intrauterine growth retardation, congenital anomalies and neonatal or postnatal pathologies of different severity. The aim of this study was to investigate the prevalence of CMV infection in a court of newborns clinically evaluated in the South Sardinia University Hospital. METHODS/RESULTS The postnatal diagnosis of CMV infection was carried out within the first 2-3 weeks of life of the newborn through the detection of the virus from urine, blood, and saliva. From September 2016 to July 2019, a total of 297 urine samples from the Department of Neonatology of Policlinico of University of Cagliari were analyzed, and proviral DNA was obtained by GeneProof Phatogen Kit. Our results showed a notable decrease in CMV infection cases in newborns, from 24% to 4% DNA positives, estimated in the South Sardinia. CONCLUSIONS The National Institute of Health underlines that, in Italy, the prevalence of infection is among the lowest in the literature. The results shown are in line with epidemiological data on the frequency of CMV infections. Our results suggest a low prevalence in comparison with the last years in the same region. REFERENCES • Kohmer N, Nagel A, Berger A, Enders M, Hamprecht K, Korn K, Kortenbusch M, Überla K, Rabenau HF. Laboratory diagnosis of congenital CMV infection in newborns: Impact of pre-analytic factors. J Clin Virol. 2019;115:32-