The spleen necrosis virus int gene product expressed in Escherichia coli has DNA binding activity and mediates att and U5-specific DNA multimer formation in vitro

To facilitate the in vitro study of the spleen necrosis virus (SNV) int gene product, we expressed the viral int locus in an Escherichia coli expression vector. Antiserum made against the protein produced in bacteria precipitated a 44-kDa polypeptide from virus-infected chicken embryo fibroblasts. This result is consistent with the expected size of the SNV int polypeptide. In a protein blotting assay, the expressed protein binds strongly to DNA and was able to complex nonspecifically with both single- and double-stranded DNAs containing or lacking viral sequences. However, under solution conditions favoring transient DNA unwinding, DNA binding was confined to supercoiled molecules containing either the SNV att sequence (the viral cis-acting region required for integration) or the U5 region of the long terminal repeat alone. Under these conditions of specific binding, multimeric DNA species were formed by apparent intermolecular interaction between protein-DNA complexes. These data indicate that retroviral integration may require local DNA unwinding at the att site for complex formation between the int gene product and DNA. This complex may be an intermediate in the viral DNA insertion process.     

In reply to Swain et al.: Re-evaluation of updated meta-analysis including trials RTOG 1016 and De-ESCALaTE

European Archives of Oto-Rhino-Laryngology -     

Red cell distribution width is a potent prognostic parameter for in-hospital and post-discharge mortality in hospitalized coronavirus disease 2019 patients: a registry-based cohort study on 3941 patients

AimTo investigate clinical and prognostic associations of red cell distribution width (RDW) in hospitalized coronavirus disease 2019 (COVID-19) patients.MethodsWe retrospectively analyzed the records of 3941 consecutive COVID-19 patients admitted to a tertiary-level institution from March 2020 to March 2021 who had available RDW on admission.ResultsThe median age was 74 years. The median Charlson comorbidity index (CCI) was 4. The majority of patients (84.1%) on admission presented with severe or critical COVID-19. Patients with higher RDW were significantly more likely to be older and female, to present earlier during infection, and to have higher comorbidity burden, worse functional status, and critical presentation of COVID-19 on admission. RDW was not significantly associated with C-reactive protein, occurrence of pneumonia, or need for oxygen supplementation on admission. During hospital stay, patients with higher RDW were significantly more likely to require high-flow oxygen therapy, mechanical ventilation, intensive care unit, and to experience prolonged immobilization, venous thromboembolism, bleeding, and bacterial sepsis. Thirty-day and post-hospital discharge mortality gradually increased with each rising RDW percent-point. In a series of multivariate Cox-regression models, RDW demonstrated robust prognostic properties at >14% cut-off level. This cut-off was associated with inferior 30-day and post-discharge survival independently of COVID-19 severity, age, and CCI; and with 30-day survival independently of COVID severity and established prognostic scores (CURB-65, 4C-mortality, COVID-gram and VACO-index).ConclusionRDW has a complex relationship with COVID-19-associated inflammatory state and is affected by prior comorbidities. RDW can improve the prognostication in hospitalized COVID-19 patients.

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), is a systemic infectious disease usually presenting with fever and respiratory symptoms (1). Although the most frequent serious manifestation of COVID-19 is pneumonia, the disease has been associated with cardiovascular, neurological, and gastrointestinal symptoms (2). Systemic inflammatory response mediated by high interleukin-6 concentrations induced by SARS-CoV-2 infection is associated with more severe clinical presentation, respiratory deterioration, and death (3,4). The presence of prior chronic comorbidities substantially affects the survival of COVID-19 patients (1).Anisocytosis, ie, unequal red blood cells (RBC) size, is a sensitive marker of distress in erythropoiesis or RBC destruction. It can be induced by various metabolic and inflammatory stimuli, nutrient deficiencies, infections, spleen disorders, and specific drugs interfering with RBC production (5). Anisocytosis can be quantified as a coefficient of variation of mean cell volume termed red blood cell distribution width (RDW), which is obtained by automatic cell counters. Higher RDW levels have recently gained attention as they are uniformly associated with unfavorable presentation and inferior outcomes in many chronic metabolic and malignant diseases (6-12). More severe clinical presentation and higher mortality rates were also found in COVID-19 patients with higher RDW levels (13-16). However, an association of RDW with other clinical outcomes in hospitalized COVID-19 patients, as well as the relationship with increased mortality in the context of other established prognostic scores, are not well defined. Thus, we aimed to investigate clinical and prognostic significance of RDW in a large cohort of hospitalized COVID-19 patients from our institution. We hypothesized that RDW was associated with more severe COVID-19 on admission and higher death rate.     

Early results after minimal lobectomy for benign nodular goiter

Following traditional operations (enucleation and subtotal resection) for benign nodular goiters recurrences may develop. Reoperations for this condition can cause complications, such as hypoparathyroidism and damage of the recurrent laryngeal nerve. That is why total thyroidectomy is recommended by many specialists in benign thyroid disease. We performed lobectomy on one side, and, if necessary, partial resection on the other side. Lobectomy was performed on one side in 31 cases, with partial resections on the other side in 73 patients. We tried to identify the parathyroid glands and both recurrent laryngeal nerves. Postoperative complications were evaluated. Temporary dysfunction of the recurrent laryngeal nerve was detected in 2.3%, permanent damage in 1.1%. Temporary hypocalcaemia developed in 16.4%, permanent hypocalcemia in 1.9%. Two reoperations were necessary for bleeding. Because of the low postoperative complication rate we recommend this method as an alternative to thyroidectomy for benign nodular goiters. We know that our favourable results can be compared with traditional subtotal resection and thyroidectomy when long term results of thyroid function and data about nodular recurrences will be collected.     

Bone mineral density variations along the lumbosacral spine

Vertebral bone quality is essential in vertebral strength and the stability of spinal screw fixation. A major factor of bone quality is the density of bone mineral and its distribution throughout the bone. There have been studies regarding bone mineral density variations in the cervical and thoracolumbar spine, but bone mineral density variations in the lumbosacral spine have not been documented. The purpose of the current study was to quantify bone mineral density variations at different lumbosacral levels and within vertebra, especially along the pathways of lumbar pedicle screws. Bone mineral density variations within the vertebrae along the lumbosacral spine were measured in 13 specimens from young male cadavers using peripheral quantitative computed tomography. Measurements included bone mineral density variations at different lumbosacral levels and in transverse layers and vertical columns within each vertebral body. These original data showed that the bone mineral density increased gradually from lumbar to S1 vertebrae, possibly reflecting a caudally increasing load on different lumbosacral levels. The highest bone mineral density in the lumbosacral spine is found at the pedicles and regions closest to pedicle bases, supporting the use of pedicle screw fixation.     

Immunogenicity of an AAV-Based COVID-19 Vaccine in Murine Models of Obesity and Aging

The SARS-CoV-2 pandemic has had a disastrous impact on global health. Although some vaccine candidates have been effective in combating SARS-CoV-2, logistical, economical, and sociological aspects still limit vaccine access globally. Recently, we reported on two room-temperature stable AAV-based COVID-19 vaccines that induced potent and protective immunogenicity following a single injection in murine and primate models. Obesity and old age are associated with increased mortality in COVID-19, as well as reduced immunogenicity and efficacy of vaccines. Here, we investigated the effectiveness of the AAVCOVID vaccine candidates in murine models of obesity and aging. Results demonstrate that obesity did not significantly alter the immunogenicity of either vaccine candidate. In aged mice, vaccine immunogenicity was impaired. These results suggest that AAV-based vaccines may have limitations in older populations and may be equally applicable in obese and non-obese populations.