Different Strategies for the Identification of SARS-CoV-2 Variants in the Laboratory Practice.

A considerable effort has been devoted in all countries to react to the COVID-19 pandemic by tracing infected individuals, containing the spread of the disease, identifying therapies, and producing and distributing vaccines. Currently, a significant concern is the appearance of variants of the virus that may frustrate these efforts by showing increased transmissibility, increased disease severity, reduced response to therapy or vaccines, and ability to escape diagnosis. All countries have therefore devoted a massive attempt to the identification and tracking of these variants, which requires a vast technological effort to sequence a large number of viral genomes. In this paper, we report our experience as one of the Italian laboratories involved in SARS-CoV-2 variant tracing. We summarize the different approaches used, and outline a potential model combining several techniques to increase tracing ability while at the same time minimizing costs.

Two-stage liver transplantation with temporary porto-middle hepatic vein shunt.

Two-stage liver transplantation (LT) has been reported for cases of fulminant liver failure that can lead to toxic hepatic syndrome, or massive hemorrhages resulting in uncontrollable bleeding. Technically, the first stage of the procedure consists of a total hepatectomy with preservation of the recipient's inferior vena cava (IVC), followed by the creation of a temporary end-to-side porto-caval shunt (TPCS). The second stage consists of removing the TPCS and implanting a liver graft when one becomes available. We report a case of a two-stage total hepatectomy and LT in which a temporary end-to-end anastomosis between the portal vein and the middle hepatic vein (TPMHV) was performed as an alternative to the classic end-to-end TPCS. The creation of a TPMHV proved technically feasible and showed some advantages compared to the standard TPCS. In cases in which a two-stage LT with side-to-side caval reconstruction is utilized, TPMHV can be considered as a safe and effective alternative to standard TPCS.

Pre-excimer laser and post-excimer laser refractive surgery measurements of scotopic pupil diameter using 2 pupillometers.

PURPOSE: To compare a digital infrared pupillometer with a handheld light amplification pupillometer for measuring scotopic pupil size and to evaluate if the postoperative refractive changes of the cornea can influence pupil measurements. DESIGN: Prospective noncomparative interventional case series. PARTICIPANTS: One hundred eyes, 50 myopic (mean spherical equivalent [SE] refraction [+/- standard deviation], -4.32+/-2.44 diopters [D]) and 50 hyperopic (mean SE refraction, +2.95+/-0.99 D), of 50 otherwise healthy subjects underwent photorefractive keratectomy or LASIK. INTERVENTION: The preoperative and postoperative scotopic pupil sizes were measured by 2 examiners (E1, E2) with both a handheld light amplification pupillometer (Colvard, Oasis Medical, Glendora, CA) and a digital infrared pupillometer (Eye World Pupillometer [EWP], Oculus Keratograph, Oculus Opikgerate GmbH, Wetzlar, Germany). The agreement and interrater repeatability were determined using the comparison method described by Bland and Altman. The paired Student's t test was used to evaluate the difference between the preoperative and postoperative measurements. MAIN OUTCOME MEASURES: Scotopic pupil diameter, topographic corneal refractive power, uncorrected visual acuity (VA), best spectacle-corrected VA, and manifest spectacle refraction. RESULTS: The preoperative mean scotopic pupil diameter was 6.12+/-0.90 mm with the EWP and 6.18+/-0.91 mm with the Colvard. After the surgery, mean SE refractions were -0.22+/-0.98 D (myopic patients) and +0.19+/-0.40 D (hyperopic patients). Postoperative mean scotopic pupil diameters were 6.12+/-0.89 mm (EWP) and 6.17+/-0.90 mm (Colvard). There was no statistically significant difference between preoperative and postoperative mean scotopic pupil sizes in either patient group. The limits of agreement between the 2 devices ranged from 2.24 mm (E1) to 2.12 mm (E2) preoperatively and from 2.27 mm (E1) to 2.08 mm (E2) postoperatively. The coefficient of interrater repeatability ranged from 0.56 mm (EWP) to 1.12 mm (Colvard) preoperatively and from 0.62 mm (EWP) to 1.14 mm (Colvard) postoperatively. CONCLUSIONS: The digital infrared pupillometer showed better preoperative and postoperative repeatability than the handheld light amplification pupillometer. In the present study, a mean correction of <3 D of the corneal refractive power did not seem to modify the preoperative scotopic pupil size measurements.

Malignancy: Changes in Circulating Immature and Mature Dendritic Cells During IL-2 Cancer Immunotherapy and Their Relation with Lymphocyte Increase and Clinical Response.

Lymphocytosis is the main biomarker predicting the efficacy of subcutaneous IL-2 anticancer immunotherapy. In addition, it has been demonstrated the fundamental role of dendritic cells (DC) in the generation of an effective anticancer immunity. However, the relation between IL-2 and DC system needs to be further understood. This preliminary study was performed in an attempt to analyze changes in circulating DC during IL-2 cancer immunotherapy in relation to lymphocyte variations and clinical efficacy of treatment. The study included 20 metastatic renal cell cancer patients, who underwent subcutaneous low-dose IL-2 immunotherapy (6.000.000 IU/day for 6 days/week for 4 weeks). To evaluate DC, venous blood samples were collected before and after 2 weeks of IL-2 injections, corresponding to the period of maximum lymphocytosis. Immature (CD123(+) ) and mature (CD11c(+) ) DC were measured by FACS and monoclonal antibodies. IL-2 induced a significant increase in the mean number of circulating mature DC, whereas no substantial change occurred in immature DC mean number. The increase in mature DC was associated with a control of disease, whereas no rise was observed in patients who had progressed on IL-2 immunotherapy. Moreover, the increase in mature DC mean number was significantly higher in patients showing evident lymphocytosis, with lymphocyte enhancement greater than 1000 cells/mmc, than in patients with less pronounced lymphocytosis, even though no significant correlation was seen in between mature DC and lymphocyte increase. This preliminary study would suggest that IL-2 may stimulate DC system and that the clinical anticancer efficacy of IL-2 is associated with the increase in circulating mature DC, which could be considered as a new favourable biomarker during IL-2 immunotherapy.