Genomic characterization of echovirus 6 causing aseptic meningitis in Hokkaido, Japan: a novel cluster in the nonstructural protein coding region of human enterovirus B

We determined four complete nucleotide sequences of echovirus 6 (E6) isolated from an epidemic of aseptic meningitis (AM) in Hokkaido, Japan, in 2011. Phylogenetic analysis of the genes encoding viral capsid protein 1 revealed that the strains were closely related to E6 strains isolated in China in recent years, but they were distantly related to E6 strains isolated from patients with AM in Osaka Prefecture, Japan, in 2011. The genes encoding the viral protease and RNA-dependent RNA polymerase (3CD) were closely related to those of several non-E6 strains of the species Human enterovirus B isolated in China, South Korea, and Australia from 1999 to 2010, resulting in a novel cluster in the phylogenetic tree. These results suggest that the incidence of AM in Japan in 2011 was caused by at least two lineages of E6 strains, and a lineage of the 3CD gene was interspersed among different serotypic strains isolated in Western Pacific countries.     

Polyvinyl alcohol scaffolds and supplementation support 3D and sphere culturing of human cancer cell lines by reducing apoptosis and promoting cellular proliferation

Three-dimensional (3D) culturing mimics the heterogeneous cellular conditions of the in vivo tumor microenvironment compared to 2D monolayer-cultured cells and 3D cultures of established cancer cell lines (sphere culture) or patient-derived cancer cells (organoid culture) are frequently used for cancer research or drug screening and evaluation. To establish more cost and time-efficient 3D culture methods for cancer cell lines, we supplemented sphere culture medium with polyvinyl alcohol (PVA) and found that 3D sphere cultures of breast and pancreatic cancer cell lines were significantly increased. Mechanistically, we found that PVA prevented cell death and promoted cellular proliferation while maintaining levels of stemness-related gene expression. Furthermore, we showed that polyvinyl formal resin (PVF) 3D scaffolds made by cross-linked PVA can function in serum-free, long-term 3D cultures to support maintenance of sphere- or tumor-like cell masses for diverse cancer cell types. Taken together, we demonstrate the effectiveness of PVA and PVF in human cancer cell line culture protocols.     

Optimized scan delay for late hepatic arterial or pancreatic parenchymal phase in dynamic contrast-enhanced computed tomography with bolus-tracking method

Objectives:To determine the optimal scan delay corresponding to individual hemodynamic status for pancreatic parenchymal phase in dynamic contrast-enhanced CT of the abdomen.Methods:One hundred and fourteen patients were included in this retrospective study (69 males and 45 females; mean age, 67.9 ± 12.1 years; range, 39–87 years). These patients underwent abdominal dynamic contrast-enhanced CT between November 2019 and May 2020. We calculated and recorded the time from contrast material injection to the bolus-tracking trigger of 100 Hounsfield unit (HU) at the abdominal aorta (s) (Time_TRIG) and scan delay from the bolus-tracking trigger to the initiation of pancreatic parenchymal phase scanning (s) (Time_SD). The scan delay ratio (SDR) was defined by dividing the Time_SD by Time_TRIG. Non-linear regression analysis was conducted to assess the association between CT number of the pancreas and SDR and to reveal the optimal SDR, which was ≥120 HU in pancreatic parenchyma.Results:The non-linear regression analysis showed a significant association between CT number of the pancreas and the SDR (p < 0.001). The mean Time_TRIG and Time_SD were 16.1 s and 16.8 s, respectively. The SDR to peak enhancement of the pancreas (123.5 HU) was 1.00. An SDR between 0.89 and 1.18 shows an appropriate enhancement of the pancreas (≥120 HU).Conclusion:The CT number of the pancreas peaked at an SDR of 1.00, which means Time_SD should be approximately the same as Time_TRIG to obtain appropriate pancreatic parenchymal phase images in dynamic contrast-enhanced CT with bolus-tracking method.Advances in knowledge:The hemodynamic state is different in each patient; therefore, scan delay from the bolus-tracking trigger should also vary based on the time from contrast material injection to the bolus-tracking trigger. This is necessary to obtain appropriate late hepatic arterial or pancreatic parenchymal phase images in dynamic contrast-enhanced CT of the abdomen.     

B-mode ultrasonographic diagnosis in gallbladder wall thickening

Diseases associated with gallbladder wall thickening include benign entities such as adenomyomatosis of the gallbladder, acute and chronic cholecystitis, and hyperplasia associated with pancreaticobiliary maljunction, and also cancer. Unique conditions such as sclerosing cholecystitis and cholecystitis associated with immune checkpoint inhibitor treatment can also manifest as wall thickening, as in some systemic inflammatory conditions. Gallbladder cancer, the most serious disease that can show wall thickening, can be difficult to diagnose early and to distinguish from benign causes of wall thickening, contributing to a poor prognosis. Differentiating between xanthogranulomatous cholecystitis and gallbladder cancer with wall thickening can be particularly problematic. Cancers that thicken the wall while coexisting with benign lesions that cause wall thickening represent another potential pitfall. In contrast, some benign gallbladder lesions that can cause wall thickening, such as adenomyomatosis and acute cholecystitis, typically show characteristic ultrasonographic features that, together with clinical findings, permit easier diagnosis. In this review of the literature, we describe B-mode abdominal ultrasonographic diagnosis of gallbladder lesions showing wall thickening.

     

Mother-to-offspring transmission of human T cell leukemia virus type I in rabbits

We studied the mode of natural transmission of human T-cell leukemia virus type I (HTLV-I) in rabbits. Four virus-infected rabbits (2 males and 2 females) were individually mated with 4 noninfected rabbits. Two virus-infected females mated with noninfected males gave birth to 7 offspring, and 2 noninfected females mated with infected males delivered 5 offspring. Four of the seven offspring born to the virus- infected mothers seroconverted for HTLV-I when aged 6 to 13 weeks with antibody titers of 1:40 to 1:160. None of the five offspring born to the noninfected mothers became seropositive during the observation period of 6 months, however. Peripheral lymphocytes were cultured with T cell growth factor, and HTLV-I-carrying lymphoid cell lines were established from the four seroconverted rabbits. All four cell lines were of T cells positive for Ia antigens. In addition, none of five newborn rabbits killed immediately after birth to a virus-infected rabbit was infected with HTLV-I. These findings provide an experimental support for the milkborne transmission of HTLV-I from mother to child in humans and indicate that the virus is tropic for T cells in rabbits as well.     

Endothelin blocks ATP-sensitive K+ channels and depolarizes smooth muscle cells of porcine coronary artery.

ATP-sensitive K+ channels with a conductance of 30 pS in smooth muscle cells of porcine coronary artery were found to be highly active in the intact cell-attached patch configuration when the pipette contained a physiological concentration of Ca2+ (greater than 10(-4) M). In the inside-out configuration, these channels were activated by extracellular Ca2+ and blocked by cytosolic ATP and glibenclamide. Endothelin applied to the pipette specifically blocked these channels in a concentration-dependent manner in the cell-attached configuration (half-maximal inhibition, 1.3 x 10(-9) M). A K+ channel opener, nicorandil, activated these channels even in the presence of 10(-8) M endothelin. In the whole-cell current-clamp method, the cell membrane was depolarized by endothelin and then repolarized by nicorandil. The membrane depolarization is closely related to contraction of smooth muscle cells. These results suggest that the ATP-sensitive K+ channels are important in controlling the vascular tone of the coronary artery and that endothelin can increase vascular tone by blocking these channels.     

S-100 beta positive T cell leukemia

We reported a peculiar case with T cell leukemia. The patient was a 34- year-old woman showing extensive splenomegaly and marked leukemic cell proliferation and running a rapid fatal clinical course. The leukemic cells were morphologically ordinary lymphocytes showing suppressor/cytotoxic(s/c) T cell phenotypes and containing S-100b protein. Southern blot analysis revealed rearrangement of the beta chain genes of the T cell receptor (TcR) of the leukemic cells. Because these phenotypic and morphologic features were identical with those of S-100 beta+T lymphocytes (S-100 beta +TL) in normal human peripheral blood, we regarded this case as S-100 beta +T cell leukemia. We discussed clinicopathological features of S-100 beta +T cell leukemia/lymphoma by assessing similar cases reported so far. S-100 beta +T cell leukemia/lymphoma is a new type of s/c T lymphocytic leukemia/lymphoma with aggressive features.