Evaluation of the Factors Associated with Reinfections towards SARS-CoV-2 Using a Case Control Design.

OBJECTIVE: The risk of reinfection with SARS-CoV-2 has been rapidly increased with the circulation of concerns about variants. So, the aim of our study was to evaluate the factors that increase the risk of this reinfection in healthcare workers compared to those who have never been positive and those who have had only one positivity. METHODS: A case-control study was carried out at the Teaching Hospital Policlinico Umberto I in Rome, Sapienza University of Rome, in the period between 6 March 2020 and 3 June 2022. Cases are healthcare workers who have developed a reinfection with the SARS-CoV-2 virus, while controls were either healthcare workers who tested positive once or those who have never tested positive for SARS-CoV-2. RESULTS: 134 cases and 267 controls were recruited. Female gender is associated with a higher odds of developing reinfection (OR: 2.42; 95% CI: 1.38-4.25). Moreover, moderate or high alcohol consumption is associated with higher odds of reinfection (OR: 1.49; 95% CI: 1.19-1.87). Diabetes is also associated with higher odds of reinfection (OR: 3.45; 95% CI: 1.41-8.46). Finally, subjects with increased red blood cell counts have higher odds of reinfection (OR: 1.69; 95% CI: 1.21-2.25). CONCLUSION: From the prevention point of view, these findings indicate that particular attention should be paid to subjects with diabetes mellitus, women and alcoholic drinkers. These results could also suggest that contact tracing represents a fundamental approach model against the SARS-CoV-2 pandemic, together with the health information of participants.

The Synergistic Effect of Time of Exposure, Distance and No Use of Personal Protective Equipment in the Determination of SARS-CoV-2 Infection: Results of a Contact Tracing Follow-Up Study in Healthcare Workers.

The aim of this study is to assess the effect of contact time, contact distance and the use of personal protective equipment on the determination of SARS-CoV-2 infection in healthcare workers (HCWs). This study consists of an analysis of data gathered for safety reasons at the Sapienza Teaching Hospital Policlinico Umberto I in Rome through the surveillance system that was put into place after the worsening of the COVID-19 pandemic. The studied subjects consist of HCWs who were put under health surveillance, i.e., all employees who were in contact with subjects who were confirmed to have tested positive for SARS-CoV-2. The HCWs under surveillance were monitored for a period encompassing ten days after the date of contact, during which they undertook nasopharyngeal swab tests analysed through RT-PCR (RealStar® SARS-CoV-2 Altona Diagnostic-Germany). Descriptive and univariate analyses have been undertaken, considering the following as risk factors: (a) no personal protective equipment use (PPE); (b) Distance < 1 m between the positive and contact persons; (c) contact time > 15'. Finally, a Cox regression and an analysis of the level of synergism between factors, as specified by Rothman, were carried out. We analysed data from 1273 HCWs. Of these HCWs, 799 (62.8%) were females, with a sample average age of 47.8 years. Thirty-nine (3.1%) tested positive during surveillance. The overall incidence rate was 0.4 per 100 person-days. Time elapsed from the last exposure and a positive RT-PCR result ranged from 2 to 17 days (mean = 7, median = 6 days). In the univariate analysis, a distance <1 m and a contact time > 15' proved to be risk factors for the SARS-CoV-2 infection, with a hazard ratio (HR) of 2.62 (95% CI: 1.11-6.19) and 3.59 (95% IC: 1.57-8.21), respectively. The synergism analysis found the highest synergism between the "no PPE use" x "Contact time". The synergy index S remains strongly positive also in the analysis of the factors "no PPE use" x "Distance" and "Time of contact" x "Distance". This study confirms the absolute need to implement safety protocols during the pandemic and to use the correct PPE within health facilities in order to prevent SARS-CoV-2 infection. The analysis shows that among the factors considered (contact time and distance, no use of PPE), there is a strong synergistic effect.

Anosmia and Ageusia as Predictive Signs of COVID-19 in Healthcare Workers in Italy: A Prospective Case-Control Study.

BACKGROUND: The aim of this study was to investigate the diagnostic accuracy of symptoms and signs in healthcare workers (HCW) with Sars-CoV-2. METHODS: This was a case-control study. Cases consisted of symptomatic healthcare workers who had a positive SARS-CoV-2 real-time polymerase chain reaction (RT-PCR) test, while controls were symptomatic healthcare workers with a negative RT-PCR test. For each symptom, ROCs were plotted. Diagnostic accuracy was calculated using the sensitivity, specificity, and positive and negative predictive values. A logistic regression analysis was carried out for calculating the OR (95% CI) for each symptom associated to the SARS-CoV-2 positivity. RESULTS: We recruited 30 cases and 75 controls. Fever had the best sensitivity while dyspnea, anosmia, and ageusia had the highest specificity. The highest PPVs were found again for dyspnea (75%), anosmia (73.7%), and ageusia (66.7%). Lastly, the highest NPVs were related to anosmia (81.4%) and ageusia (79.3%). Anosmia (OR = 14.75; 95% CI: 4.27-50.87), ageusia (OR = 9.18; 95% CI: 2.80-30.15), and headache (OR = 3.92; 95% CI: 1.45-10.56) are significantly associated to SARS-CoV-2 positivity. CONCLUSIONS: Anosmia and ageusia should be considered in addition to the well-established fever, cough, and dyspnea. In a resource-limited setting, this method could save time and money.

Diagnostic performance in active TB of QFT-Plus assay and co-expression of CD25/CD134 in response to new antigens of Mycobacterium tuberculosis.

The new QuantiFERON-TB Gold Plus employs modified peptides optimized to elicit an IFNγ response from CD8+ cytotoxic T lymphocytes in addition to CD4+ T cells. With a view to improve the difficult identification of TB cases, we assessed the combination of two specific immunological markers comprising IFNγ secretion and T cells co-expression of CD25 and CD134 in response to Mycobacterium tuberculosis-specific antigens. A total of 34 subjects with suspected TB and 10 age-matched HD were prospectively enrolled. Assessing the performance of QFT-Plus in terms of the TB1 and TB2 results, we found that in TB patients, the quantitative IFNγ value in TB2 was similar to that in TB1, and we did not find any differences irrespective of the disease (pulmonary or extra-pulmonary). The flow cytometric CD25/CD134 assay, allowed a more accurate differentiation between M. tuberculosis-infected and uninfected patients, with a better combination of sensitivity and specificity, especially by evaluation of CD4+ T-cell subset. All individuals with negative QFT-Plus results displayed a positive CD25/CD134 response. Overall, a positive correlation was found between T cells co-expressing CD25/CD134 and IFNγ levels in response to both QFT-Plus TB antigen tubes, as well as between the QFT-Plus TB1 and TB2 tubes. We demonstrated that both TB1 and TB2 induce a higher expression of CD25+CD134+ markers on CD4+ T cells among infected TB subjects, compared to the lower degree of CD8+ T cells, mainly induced to TB2 stimulation. We suggest that a combined use of classic QFT-Plus and specific CD25/CD134 response may be a useful means in the diagnostic workup for active TB.

Myeloid and lymphoid activation markers in AIDS and non-AIDS presenters.

HIV infection is characterized by a state of chronic activation of the immune system, which is not completely reversed by antiretroviral treatment (ART). The aim of this study was to assess myeloid and lymphoid activation markers during HIV infection, before and one year after ART initiation, in AIDS and non-AIDS presenters. Treatment naïve HIV positive patients were enrolled in this study. Myeloid dendritic cell (mDC), plasmacytoid dendritic cell (pDC), slanDC, monocyte and T-lymphocyte cell counts and activation status, were assessed by flow cytometry in peripheral blood samples. Soluble (s)CD14 and sCD163 were assessed in plasma samples using ELISA assays. Statistical analyses were performed using GraphPad Prism and Minitab Express. Thirty-four ART naïve HIV-1 infected subjects were enrolled in this study (22 non-AIDS and 12 AIDS presenters). Seventeen healthy donors (HD) were included as control group. Although circulating mDC levels resulted unchanged, HLA-DR expression was decreased on mDCs of HIV positive subjects compared to HD (p < 0,0001). AIDS presenters showed the lowest level of expression of HLA-DR on mDCs. Circulating levels of pDCs were decreased in HIV patients compared to HD (p < 0,001), without any changes in HLA-DR expression. SlanDC cell counts were extremely reduced in AIDS presenters, compared to non-AIDS presenters and HD (p < 0,01 and p < 0,0001, respectively) and showed higher HLA-DR expression in HIV patients compared to HD (p < 0,01). Intermediate monocyte (IM) cell counts were increased in AIDS and non-AIDS presenters compared to HD (p < 0,001 and p < 0,001 respectively). Furthermore, IM expansion was directly correlated to HIV viral load (p = 0,036) and independent from CD4 cell counts and activation levels. Plasma concentrations of sCD14 and sCD163 resulted increased in HIV infected subjects compared to HD (p < 0,0001 and p < 0,001), with the highest levels observed in AIDS presenters. After 1 year, ART was able to increase pDC and decrease IM absolute cell counts and modify HLA-DR expression on mDCs and slanDCs, approaching the levels observed in HD. ART reduced also CD4 and CD8 activation levels. In conclusion, in untreated HIV infected subjects circulating dendritic cells resulted altered either in numbers or in HLA-DR expression, especially in AIDS presenters. IM absolute counts were equally increased in AIDS and non-AIDS presenters. ART was able to reduce myeloid and lymphoid inflammation in both advanced and non-advanced HIV patients, confirming the role of ART in hampering disease progression and immune activation associated non-AIDS events.

Intracranial tuberculous mass lesions treated with thalidomide in an immunocompetent child from a low tuberculosis endemic country: A case report.

RATIONALE: Tuberculous meningitis is a highly morbid, often fatal disease. PATIENT CONCERN: We describe a case of an Italian child. DIAGNOSES:: we diagnosed early a Tuberculous meningitis complicated by the occurrence of hydrocephalus, stroke, and paradoxical reaction with brain pseudo-abscesses. INTERVENTIONS: The child started readily a specific therapy associated with steroids and thalidomide was introduced few month later. OUTCOMES: the patient had a favorable outcome without neurologic sequelae. LESSONS: Despite the prompt specific anti-tubercular and adjuvant corticosteroid therapies, only the addition of thalidomide to the treatment allow to a favorable clinical outcome.

The role of IL-15 in challenging Acquired Immunodeficiency Syndrome.

OBJECTIVE: To determine the functions of in vitro primed Natural Killer (NK) cells in Human Immunodeficiency Virus (HIV-1) infection and the role of IL-2, IL-12 and IL-15 in enhancing the NK survival and activity in terms of viral suppression and of purging of HIV provirus. METHODS: Peripheral Blood Mononuclear Cells (PBMCs) and CD4+ T lymphocytes cells obtained from eight healthy donors were infected in vitro with HIV-1 and p24 was measured with and without IL-2, IL-12 and IL-15. We studied the effect of NK pulsed in vitro with IL-2, IL-12 and IL-15 on HIV replication by measurement of p-24 and DNA-provirus load when added into the culture of PBMCs and CD4+ T lymphocytes cells infected in vitro. We evaluated the effect of NK cells pulsed with IL-2, IL-12 and IL-15 on HIV replication and DNA-load into the culture of CD4+ T lymphocytes cells and PBMCs by trans-well chamber. RESULTS: We found high levels of p24 in the supernatants of PBMCs and CD4+ T lymphocytes cells cultured with IL-2, IL-12, and IL-15. We observed a significant reduction of p24 in the culture both of infected PBMCs and CD4+ T lymphocytes cells in which was added NK pulsed with IL-15. We did not obtain the some results with NK pulsed with IL-2 and IL-12. We observed a power effect of NK pulsed with IL-15 on HIV-DNA. The trans-well chamber experiments showed that the effect of NK is both direct and both mediated by realizing of soluble factors. CONCLUSIONS: This study highlights some important effects of IL 15 on NK in HIV patients anyway our results are preliminary and descriptive and others studies will be needed to provide rationale for immune therapies.

Candidal thrombophlebitis of central veins: case report and review.

Although candidemia and central catheter septic thrombosis is quite common, central veins thrombophlebitis caused by Candida spp. is a rarely reported complication in critically ill patients. Here we report a case of thrombophlebitis of the right internal jugular and subclavian veins due to Candida albicans which occurred in a patient admitted in the intensive care unit for major trauma. The individual was eventually cured after prolonged course of antifungal therapy. We also review 24 additional cases of Candida induced central veins thrombophlebitis reported since 1978. A central vein catheter was in place in all 25 patients with 21 (84%) being admitted in an intensive care unit, 22 (88%) were receiving total parenteral nutrition and 23 (92%) undergoing a course of antibiotic therapy. Overall mortality was 16%, including two patients who received no therapy and died. In the group of patients receiving only medical therapy, the mortality rate was 13%, while no deaths were observed among those treated with combined medical and surgical therapy. Literature data suggest that Candida caused central veins thrombophlebitis is a rare and probably underdiagnosed infectious complication of the critically ill patient. Despite the dramatic presentation with persistent candidemia, mortality is low even with a conservative medical approach with prolonged fungicidal therapy through the use of amphotericin B or echinocandins. Thus, the decision for a combined surgical debridement should be assessed for each patient.

Invasive aspergillosis in patients with liver disease.

Invasive aspergillosis (IA) has been traditionally considered an infection occurring in patients with well established risk factors, such as neutropenia, hematologic malignancies, organ transplantation, or HIV. However there is increasing evidence that apparently immunocompetent patients, such as those with severe liver disease, are also at high risk for Aspergillus infections. Here we report two cases of proven invasive aspergillosis and review 72 others of aspergillosis reported since 1973 in patients with liver disease. Most patients had end-stage cirrhosis or acute hepatic failure. Overall mortality rate was 72.2% and the majority of patients who died had CNS involvement, disseminated infections, and received antifungal agents on a less common basis. A trend toward higher survival for cases reported during the period 2000-2009 was observed. Literature data suggest that invasive aspergillosis is a potential fatal complication of severe liver disease. The high mortality rate observed in these patients appears to be related not only to the severity of their underlying conditions, but also to a lack in clinical diagnosis. New diagnostic tools, e.g., galactomannan (GM) antigen test, in association with increased clinical suspicion may allow an early diagnosis and improve the outcome of IA in this particular category of patients.

Mutational resistance pattern of HIV type 1 in CD14+ monocytes, CD4+ T cells, and plasma from treated patients.

It is necessary to understand the molecular nature of the virus population that persists in cellular reservoirs. To achieve this we planned to characterize the patterns of resistance of HIV-1 in CD14(+) monocytes, CD4(+) T cells, and plasma. Blood samples were collected from 42 patients treated for HIV: 32 were in virological failure and in 10 viremia was undetectable. CD14(+) and CD4(+) T cells were isolated using magnetic beads. Genotyping of the reverse transcriptase and protease gene of HIV-1 was undertaken using the fluorescent dideoxy-terminator method. Of the 32 patients in virological failure, 24 (75%) had resistance mutations in at least one compartment. The numbers and types of mutations from monocytes were the same as those detected in both CD4(+) T cell-associated virus and plasma in only 8% whereas in 71% monocytes exhibited a different mutation pattern. In 21% of patients, the profile of drug-resistant mutations in the virus from blood monocytes was identical to that in plasma but differed from that in CD4. In the 71% of patients with virological suppression, the genotypic resistance pattern differed between monocytes and CD4(+) T cells. Circulating monocytes may harbor a viral dominant population different from those viruses circulating in blood and archived in CD4(+) T cells. Hence, monocytes and other cellular reservoirs might serve as an indirect source of a drug-resistant viral variant.

Primary retroperitoneal abscesses due to Rhodococcus equi in a patient with severe nephrotic syndrome: successful antibiotic treatment with linezolid and tigecycline.

We present a case of Rhodococcus equi primary retroperitoneal abscesses without pulmonary involvement in an immunocompromised patient with severe nephrotic syndrome. No risk factors for exposure to R. equi were present. The infection was successfully treated with long-term combination antibiotic treatment including linezolid and tigecycline.

In vivo and in vitro effects of antituberculosis treatment on mycobacterial interferon-gamma T cell response.

BACKGROUND: In recent years, the impact of antituberculous treatment on interferon (IFN)-gamma response to Mycobacterium tuberculosis antigens has been widely investigated, but the results have been controversial. The objective of the present study was: i) to evaluate longitudinal changes of IFN-gamma response to M. tuberculosis-specific antigens in TB patients during antituberculous treatment by using the QuantiFERON-TB Gold (QFT-G) assay; ii) to compare the differences in T-cell response after a short or prolonged period of stimulation with mycobacterial antigens; iii) to assess the CD4+ and CD8+ T cells with effector/memory and central/memory phenotype; iv) to investigate the direct in vitro effects of antituberculous drugs on the secretion of IFN-gamma. PRINCIPAL FINDINGS: 38 TB patients was evaluated at baseline and at month 2 and 4 of treatment and at month 6 (treatment completion). 27 (71%) patients had a QFT-G reversion (positive to negative) at the end of therapy, while 11 (29%) TB patients remained QFT-G positive at the end of therapy. Among the 11 patients with persistent positive QFT-G results, six had a complete response to the treatment, while the remaining 5 patients did not have a resolution of the disease. All 27 patients who became QFT-G negative had a complete clinical and microbiological recovery of the TB disease. In these patients the release of IFN-gamma is absent even after a prolonged 6-day incubation with both ESAT-6 and CFP-10 antigens and the percentage of effector/memory T-cells phenotype was markedly lower than subjects with persistent positive QFT-G results. The in vitro study showed that antituberculous drugs did not exert any inhibitory effect on IFN-gamma production within the range of therapeutically achievable concentrations. CONCLUSIONS: The present study suggests that the decrease in the M. tuberculosis-specific T cells responses following successful anti-TB therapy may have a clinical value as a supplemental tool for the monitoring of the efficacy of pharmacologic intervention for active TB. In addition, the antituberculous drugs do not have any direct down-regulatory effect on the specific IFN-gamma response.

Simplified maintenance therapy with abacavir/lamivudine/zidovudine plus tenofovir after sustained HIV load suppression: four years of follow-up.

PURPOSE: To assess the virologic and immunologic outcome of a treatment simplification strategy based on the substitution of protease inhibitor (PI)-based regimen with abacavir/lamivudine/zidovudine (ABC/3TC/ZDV, also known as trizivir or TZV) plus tenofovir (TDF) in viral-suppressed patients. METHOD: The study population included 17 HIV-infected patients with undetectable viral loads over 12 months of a stable PI-based therapy. Patients were switched to a combination of TZV (2 pills twice a day) plus TDF (1 pill once a day) and were followed up for 48 months. They were studied for intracellular HIV DNA, CD4 cell count, HIV RNA levels, and lipid metabolism. RESULTS: All patients had undetectable HIV RNA for the entire period of the follow-up. After 24 months of treatment with TZV plus TDF, the levels of cellular HIV DNA significantly decreased (p = .021). When we stratified the patients on the basis of HIV DNA outcome, we observed a significant increase of CD4 count only in patients who had undetectable HIV DNA after 24 months of TZV/TDF treatment. On the contrary, the CD4 count did not change in patients whose HIV DNA was still detectable at 24 months. The percentage of patients taking lipid-lowering agents declined significantly after switching to TZV/TDF. CONCLUSION: This small pilot study suggests that a single-class quadruple regimen of TZV/TDF may represent a safe and appealing approach in the setting of simplification/switching antiretroviral strategies.