The Establishment of a Primary Culture System of Proximal Tubule Segments Using Specific Markers from Normal Mouse Kidneys

The proximal tubule contains the highest expression of angiotensinogen mRNA and protein within the kidney and plays a vital role in the renal renin-angiotensin system. To study the regulation of angiotensinogen expression in the kidney in more detail, the proximal tubule needs to be accurately isolated from the rest of the nephron and separated into its three segments. The purpose of this study was to design a novel protocol using specific markers for the separation of proximal tubule cells into the three proximal tubule segments and to determine angiotensinogen expression in each segment. Kidneys were removed from C57BL/6J mice. The proximal tubules were aspirated from region of a Percoll gradient solution of the appropriate density. The proximal tubule was then separated into its three segments using segment-specific membrane proteins, after which each segment was characterized by a different specific marker (sodium-glucose transporter 2 for Segment 1; carbonic anhydrase IV for Segment 2; ecto-adenosine triphosphatase for Segment 3). The isolation of proximal tubules into three segments was successful, and angiotensinogen mRNA in Segment 2 and 3 and angiotensinogen protein in all three segments were confirmed. This protocol will be helpful for future studies of the detailed mechanisms of the intrarenal renin-angiotensin system

In-plane anisotropy of the single-qq and multiple-qq ordered phases in the antiferromagnetic metal CeRh2_2Si2_2 unveiled by the bulk measurements under uniaxial stress and neutron scattering

We performed magnetization, resistivity, and neutron diffraction measurements under uniaxial stress applied along [1-10] direction on the tetragonal magnet CeRh$_2$Si$_2$ with commensurate magnetic orders. CeRh$_2$Si$_2$ has two successive antiferromagnetic (AF) orders in zero magnetic field. The high temperature phase (AF1 phase) has the magnetic modulation wave vector of $q = (\frac{1}{2}, \frac{1}{2}, 0)$, and the low temperature phase (AF2 phase) is characterized by the four $q$-vectors of $q = (\frac{1}{2}, \frac{1}{2}, 0), (\frac{1}{2}, -\frac{1}{2}, 0), (\frac{1}{2}, \frac{1}{2}, \frac{1}{2})$, and $(\frac{1}{2}, -\frac{1}{2}, \frac{1}{2})$. By measuring the uniaxial stress dependence of the magnetization, resistivity and the intensities of magnetic Bragg reflections, we confirmed that the AF1 phase has the single-$q$ magnetic order with two-fold rotational symmetry and the AF2 phase has the multi-$q$ magnetic order with four-fold rotational symmetry. In order to understand the origin of multi-$q$ order of CeRh$_2$Si$_2$, we also performed inelastic neutron scattering measurement on the single crystal samples. We found a magnetic excitation at the transfer energy $\hbar \omega \sim$ 8 meV. By applying the linear spin-wave theory, we found that the nearest and the next-nearest neighbor exchange interactions on the $ab$-plane, $J_1$ and $J_2$, are dominant in the AF2 phase. However, the $J_1$-$J_2$ model cannot lift the degeneracy between the single-$q$ (AF1) and multi-$q$ (AF2) phases. We suggest that it can be lifted by taking into account the biquadratic interaction derived from the perturbative expansion for the Kondo lattice Hamiltonian. [S. Hayami et al., Phys. Rev. B 95, 224424 (2017).Comment: 7 pages, 7 figure

Risk stratification for the prognosis of patients with chemoresistant urothelial cancer treated with pembrolizumab

The use of immune checkpoint inhibitors to treat urothelial carcinoma (UC) is increasing rapidly without clear guidance for validated risk stratification. This multicenter retrospective study collected clinicopathological information on 463 patients, and 11 predefined variables were analyzed to develop a multivariate model predicting overall survival (OS). The model was validated using an independent dataset of 292 patients. Patient characteristics and outcomes were well balanced between the discovery and validation cohorts, which had median OS times of 10.2 and 12.5 mo, respectively. The final validated multivariate model was defined by risk scores based on the hazard ratios (HRs) of independent prognostic factors including performance status, site of metastasis, hemoglobin levels, and the neutrophil-to-lymphocyte ratio. The median OS times (95% confidence intervals [CIs]) for the low-, intermediate-, and high-risk groups (discovery cohort) were not yet reached (NYR) (NYR–19.1), 6.8 mo (5.8-8.9), and 2.3 mo (1.2-2.6), respectively. The HRs (95% CI) for OS in the low- and intermediate-risk groups vs the high-risk group were 0.07 (0.04-0.11) and 0.23 (0.15-0.37), respectively. The objective response rates for in the low-, intermediate-, and high-risk groups were 48.3%, 28.8%, and 10.5%, respectively. These differential outcomes were well reproduced in the validation cohort and in patients who received pembrolizumab after perioperative or first-line chemotherapy (N = 584). In conclusion, the present study developed and validated a simple prognostic model predicting the oncological outcomes of pembrolizumab-treated patients with chemoresistant UC. The model provides useful information for external validation, patient counseling, and clinical trial design

Weak glow at Tarumae volcano, Japan, witnessed by the high-sensitive camera soon after the Tokachi-oki Earthquake in 2003 (MJMA 8.0)

In this paper, the authors describe remarkable thermo-activities especially at the fumaroles B on the southwestern cliff of the summit dome on Tarumae volcano, which unusually occurred soon after the Tokachi-oki erathquake that took place on Sep. 26 2003 (MJMA 8.0). The unusual thermoactivities include (1) increase in gas flux, (2) weak glow witnessed by the high-sensitive camera in the nighttime with positions moving night by night, and (3) ash ejection of about 24m^3. Since the high-sensitive cameras can detect thermal radiation, the observed glow would be evidence for high-temperature of rock surface. It is considered that the Tokachi-oki earthquake would affect the volcano to eject a large amount of high-temperature gas, which resulted in the weak but unusual glow and ash deposits of the order of 10m^3 in volume

ATF6β is a host cellular target of the Toxoplasma gondii virulence factor ROP18

Toxoplasma virulence factor ROP18 targets endoplasmic reticulum–bound transcription factor ATF6β in the host cell, leading to the detrimental loss of ATF6β through proteasome-dependent degradation

A Single Nucleotide Polymorphism within the Acetyl-Coenzyme A Carboxylase Beta Gene Is Associated with Proteinuria in Patients with Type 2 Diabetes

It has been suggested that genetic susceptibility plays an important role in the pathogenesis of diabetic nephropathy. A large-scale genotyping analysis of gene-based single nucleotide polymorphisms (SNPs) in Japanese patients with type 2 diabetes identified the gene encoding acetyl-coenzyme A carboxylase beta (ACACB) as a candidate for a susceptibility to diabetic nephropathy; the landmark SNP was found in the intron 18 of ACACB (rs2268388: intron 18 +4139 C > T, p = 1.4×10−6, odds ratio = 1.61, 95% confidence interval [CI]: 1.33–1.96). The association of this SNP with diabetic nephropathy was examined in 9 independent studies (4 from Japan including the original study, one Singaporean, one Korean, and two European) with type 2 diabetes. One case-control study involving European patients with type 1 diabetes was included. The frequency of the T allele for SNP rs2268388 was consistently higher among patients with type 2 diabetes and proteinuria. A meta-analysis revealed that rs2268388 was significantly associated with proteinuria in Japanese patients with type 2 diabetes (p = 5.35×10−8, odds ratio = 1.61, 95% Cl: 1.35–1.91). Rs2268388 was also associated with type 2 diabetes–associated end-stage renal disease (ESRD) in European Americans (p = 6×10−4, odds ratio = 1.61, 95% Cl: 1.22–2.13). Significant association was not detected between this SNP and nephropathy in those with type 1 diabetes. A subsequent in vitro functional analysis revealed that a 29-bp DNA fragment, including rs2268388, had significant enhancer activity in cultured human renal proximal tubular epithelial cells. Fragments corresponding to the disease susceptibility allele (T) had higher enhancer activity than those of the major allele. These results suggest that ACACB is a strong candidate for conferring susceptibility for proteinuria in patients with type 2 diabetes