Mapping of SARS-CoV-2 in Waldeyer's lymphatic ring and visceral biopsies: the age and the illness duration's impact.

OBJECTIVE: To study the impact of age and the interval between disease diagnosis and death on the organotropism of SARS-CoV-2. METHOD: Patients underwent post-mortem biopsies from lungs, Waldeyer ring, heart, liver, kidneys and bone marrow between 2020‒2021. SARS-CoV-2 organotropism was mapped by using molecular RT-PCR analyses for SARS-CoV2 targeting the Envelope gene (E), the RNA Polymerase Gene (RdRp), and the Nucleocapsid gene (N). Statistical and linear regression analysis was performed to study the impact of age and illness duration in SARS-CoV-2 organotropism. RESULTS: We performed 158 postmortem biopsies in 21 patients, with a mean age of 76 years old. The mean interval between the diagnosis of the infection to the death was 23 days. The RNA of the SARS-CoV-2 was detected in 100% of lung biopsies, 76%‒82% of Waldeyer's ring biopsies, 55% of heart biopsies, 40% of kidney biopsies, 33% of liver and 25% of bone marrow biopsies. Patients who died before the day 9, presented extensive visceral dissemination of SARS-CoV-2 RNA. Most of the patients older than 80 years (90%) presented visceral dissemination of SARS-CoV-2 RNA, while among younger patients, only 3/11 patients presented visceral dissemination of the virus. The relationship between "age" and "illness duration" and multitropism of the virus was statistically significant (p<0.001). CONCLUSION: Disease interval and age were factors that were significantly associated with RT-PCR positive results in multiple organs. Critical COVID-19 patients have multiorganic viral dissemination in early stages. In the critical older patients, multiorganic viral dissemination is the rule.

Mapping of SARS-CoV-2 in Waldeyer's lymphatic ring and visceral biopsies: the age and the illness duration's impact

Abstract Objective: To study the impact of age and the interval between disease diagnosis and death on the organotropism of SARS-CoV-2. Method: Patients underwent post-mortem biopsies from lungs, Waldeyer ring, heart, liver, kidneys and bone marrow between 2020-2021. SARS-CoV-2 organotropism was mapped by using molecular RT-PCR analyses for SARS-CoV2 targeting the Envelope gene (E), the RNA Polymerase Gene (RdRp), and the Nucleocapsid gene (N). Statistical and linear regression analysis was performed to study the impact of age and illness duration in SARS-CoV-2 organotropism. Results: We performed 158 postmortem biopsies in 21 patients, with a mean age of 76 years old. The mean interval between the diagnosis of the infection to the death was 23 days. The RNA of the SARS-CoV-2 was detected in 100% of lung biopsies, 76%-82% of Waldeyer's ring biopsies, 55% of heart biopsies, 40% of kidney biopsies, 33% of liver and 25% of bone marrow biopsies. Patients who died before the day 9, presented extensive visceral dissemination of SARS-CoV-2 RNA. Most of the patients older than 80 years (90%) presented visceral dissemination of SARS-CoV-2 RNA, while among younger patients, only 3/11 patients presented visceral dissemination of the virus. The relationship between "age" and "illness duration" and multitropism of the virus was statistically significant (p< 0.001). Conclusion: Disease interval and age were factors that were significantly associated with RT-PCR positive results in multiple organs. Critical COVID-19 patients have multiorganic viral dissemination in early stages. In the critical older patients, multiorganic viral dissemination is the rule. Level of evidence: 4. Case Series.

Cranioplasty with Autologous Bone Flaps Cryopreserved with Dimethylsulphoxide: Does Tissue Processing Matter.

OBJECTIVE: The aim of this article was to study the outcome of patients who underwent cranioplasty with cryopreserved autologous bone after decompressive craniectomy. METHODS: Data from 74 patients were retrospectively analyzed. They were divided into groups according to the storage time and the age at cranioplasty. To assess the predictive potential for complication, factors were related to successive stages (preoperative, craniectomy, tissue processing, cranioplasty, and postoperative). Cooling and warming rates applied on bone flap were calculated. The ability to inhibit microbial growth was determined exposing bone fragments to a panel of microorganisms. The concentration of antibiotics eluted from the bone was also determined. A bone explant culture method was used to detect living cells in the thawed cranial bone. RESULTS: Hydrocephalus was significantly more frequent in pediatric patients (26.7%) than in adults (5.1%). The overall rate of bone flap resorption was 21.6% (43.7% of which required reoperation). Surgical site infection after cranioplasty was detected in 6.8% of patients. There was no correlation between infection as a postoperative complication and previous microbiological-positive culture during processing. The cause of craniectomy did not influence the risk of bone flap contamination. Vancomycin was the only antibiotic detected in the supernatant where the bone was incubated. Outgrowth from bone explants was observed in 36.8% of thawed skulls. An early start of bone flap processing at the tissue bank had a positive effect on cell viability. CONCLUSIONS: The outcome after autologous cranioplasty is a multifactorial process, which is modulated by patient-related, surgery-related, and bone-related factors.

The molecular epidemiology of HIV-1 in the Comunidad Valenciana (Spain): analysis of transmission clusters.

HIV infections are still a very serious concern for public heath worldwide. We have applied molecular evolution methods to study the HIV-1 epidemics in the Comunidad Valenciana (CV, Spain) from a public health surveillance perspective. For this, we analysed 1804 HIV-1 sequences comprising protease and reverse transcriptase (PR/RT) coding regions, sampled between 2004 and 2014. These sequences were subtyped and subjected to phylogenetic analyses in order to detect transmission clusters. In addition, univariate and multinomial comparisons were performed to detect epidemiological differences between HIV-1 subtypes, and risk groups. The HIV epidemic in the CV is dominated by subtype B infections among local men who have sex with men (MSM). 270 transmission clusters were identified (>57% of the dataset), 12 of which included ≥10 patients; 11 of subtype B (9 affecting MSMs) and one (n = 21) of CRF14, affecting predominately intravenous drug users (IDUs). Dated phylogenies revealed these large clusters to have originated from the mid-80s to the early 00 s. Subtype B is more likely to form transmission clusters than non-B variants and MSMs to cluster than other risk groups. Multinomial analyses revealed an association between non-B variants, which are not established in the local population yet, and different foreign groups.

Use of liquid nitrogen during storage in a cell and tissue bank: contamination risk and effect on the detectability of potential viral contaminants.

Cryopreservation is widely used for banking cells and tissues intended for transplantation. Liquid nitrogen provides a very stable ultra-low temperature environment. Thus, it is used for longterm storage. Unlike the exhaustive microbiological monitoring of the environmental conditions during tissue processing, storage is not usually considered as a critical point of potential contamination risk in professional standards for cell and tissue banking. We have analysed the presence of microbial agents inside our nitrogen tanks. We have mainly detected environmental and water-borne bacteria and fungi. In addition, we have studied the effect of liquid nitrogen exposure on virus detectability. Only differences for hepatitis C virus RNA were observed. Measures for contamination risk reduction during storage must be mandatory in cell and tissue banking.

Comparison of BinaxNOW Influenza A&B assay and real-time reverse transcription polymerase chain reaction for diagnosis of influenza A pandemic (H1N1) 2009 virus infection in adult patients.

The BinaxNOW Influenza A&B assay was evaluated for the diagnosis of influenza A pandemic (H1N1) 2009 virus infection in 354 adult patients. The sensitivity, specificity, and positive and negative predictive values were 32%, 100%, 100%, and 67%, respectively. The analytic sensitivity of the assay was log105.0 tissue culture infective dose (TCID)50/mL.

Sensitivity of a marketed immunochromatographic assay specifically targeting the pandemic influenza A/H1N1 2009 virus.

The sensitivity of an immunochromatographic assay specifically detecting the pandemic influenza A/H1N1 2009 virus (influenza Ag A/B/A(H1N1) pandemic rapid test; SD Standard Diagnostics, South Korea) was evaluated. The overall sensitivity of the assay and its analytic sensitivity were 26.9% and 2.6 x 10(4) TCID(50)/mL, respectively.

Impact of cardiovascular risk factors on oxidative stress and DNA damage in a high risk Mediterranean population.

The impact of classic cardiovascular risk factors on oxidative stress status in a high-risk cardiovascular Mediterranean population of 527 subjects was estimated. Oxidative stress markers (malondialdehyde, 8-oxo-7'8'-dihydro-2'-deoxyguanosine, oxidized/reduced glutathione ratio) together with the activity of antioxidant enzyme triad (superoxide dismutase, catalase, glutathione peroxidase) were analysed in circulating mononuclear blood cells. Malondialdehyde, oxidized glutathione and the ratio of oxidized to reduced glutathione were significantly higher while catalase and glutathione peroxidase activities were significantly lower in high cardiovascular risk participants than in controls. Statistically significant differences were obtained after additional multivariate control for sex, age, obesity, diabetes, lipids and medications. Among the main cardiovascular risk factors, hypertension was the strongest determinant of oxidative stress in high risk subjects studied at a primary prevention stage.