X-ray TV system for measuring microcirculation in small pulmonary vessels

We developed a new system that consists of 1) a specially designed X-ray apparatus, 2) an X-ray-sensitive 1-in. Vidicon camera, and 3) a digital image-processing device. The picture element is approximately 20 micron in size, and the time required for one frame is 1/30 s. Using this system, we measured the internal diameter (ID), the cross-sectional area, flow velocity, volume flow, and transit time of small pulmonary vessels of approximately 100-500 micron at control and with serotonin in anesthetized cats. Flow velocity and volume flow from large [458 +/- 22 (SE) micron] to small (340 +/- 32 micron) arteries were 5.4 +/- 0.4 cm/s and 0.53 +/- 0.06 ml/min, respectively. Transit times of the contrast medium from large to small arteries (Ta) and to large veins (Tv) were 0.68 +/- 0.04 and 3.71 +/- 0.25 s, respectively. Serotonin injection (20-30 micrograms/kg iv) decreased ID, flow velocity, and volume flow of arteries by 8-48, 32, and 76%, respectively, whereas Ta and Tv increased by 91 and 69%, respectively. The system can provide useful information regarding the local circulation in the lung.     

Classification and characteristics of Japanese patients with antineutrophil cytoplasmic antibody-associated vasculitis in a nationwide,prospective, inception cohort study

Introduction

We investigated the clinical and serological features of patients with antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) in Japan using data from a nationwide, prospective, inception cohort study.

Methods

In total, 156 Japanese patients with newly diagnosed AAV were classified according to the European Medicines Agency (EMEA) algorithm with exploratory surrogate markers for AAV-related non-granulomatous pulmonary lesions, predefined as alveolar haemorrhage and interstitial lung disease (ILD), and their clinical and serological features were evaluated.

Results

Using the EMEA algorithm, we identified 14 patients (9.0%) with eosinophilic granulomatosis with polyangiitis (EGPA), 33 (21.2%) with granulomatosis with polyangiitis (GPA), 78 (50.0%) with microscopic polyangiitis and renal-limited vasculitis (MPA/RLV), and 31 (19.9%) with unclassifiable vasculitis. The average ages of patients with EGPA (male/female, 5/9), GPA (12/21), and MPA/RLV (35/43) and unclassifiable (9/22) were 58.0, 63.6, 71.1, and 70.6 years, respectively. Myeloperoxidase (MPO)-ANCA and proteinase-3 ANCA positivity was 50.0% and 0% for EGPA, 54.6% and 45.5% for GPA, 97.4% and 2.6% for MPA/RLV, and 93.5% and 3.2% for unclassifiable, respectively. According to the Birmingham Vasculitis Activity Score (BVAS), cutaneous (71.4%) and nervous system (92.9%) manifestations were prominent in EGPA and ear, nose, and throat manifestations (84.9%) and chest manifestations (66.7%) in GPA. Renal manifestations developed frequently in MPA/RLV (91.0%) and GPA (63.6%). The average serum creatinine levels were 0.71 mg/dL for EGPA, 1.51 mg/dL for GPA, 2.46 mg/dL for MPA/RLV, and 0.69 mg/dL for unclassifiable. The percentages of patients with ILD were 14.3% for EGPA, 9.0% for GPA, 47.4% for MPA/RLV, and 61.3% for unclassifiable. Patients with ILD (n = 61) had significantly lower BVAS (P = 0.019) with fewer ear, nose, and throat and cardiovascular manifestations than patients without ILD (n = 95).

Conclusions

MPO-ANCA-positive MPA/RLV is the most common form of AAV in Japanese patients, and one-half of patients with GPA were positive for MPO-ANCA. ILD is an important clinical manifestation in Japanese patients with AAV. Unclassifiable vasculitis with MPO-ANCA positivity and ILD may represent a novel variant of MPA.

Trial Registration

The University Hospital Medical Information Network Clinical Trials Registry: UMIN000001648. Registered 28 February 2009.     

Differential expression of Toll-like receptors on human alveolar macrophages and autologous peripheral monocytes

Background

Toll-like receptors (TLRs) are critical components in the regulation of pulmonary immune responses and the recognition of respiratory pathogens such as Mycobacterium Tuberculosis (M.tb). Through examination of human alveolar macrophages this study attempts to better define the expression profiles of TLR2, TLR4 and TLR9 in the human lung compartment which are as yet still poorly defined.

Methods

Sixteen healthy subjects underwent venipuncture, and eleven subjects underwent additional bronchoalveolar lavage to obtain peripheral blood mononuclear and bronchoalveolar cells, respectively. Surface and intracellular expression of TLRs was assessed by fluorescence-activated cell sorting and qRT-PCR. Cells were stimulated with TLR-specific ligands and cytokine production assessed by ELISA and cytokine bead array.

Results

Surface expression of TLR2 was significantly lower on alveolar macrophages than on blood monocytes (1.2 ± 0.4% vs. 57 ± 11.1%, relative mean fluorescence intensity [rMFI]: 0.9 ± 0.1 vs. 3.2 ± 0.1, p < 0.05). The proportion of TLR4 and TLR9-expressing cells and the rMFIs of TLR4 were comparable between alveolar macrophages and monocytes. The surface expression of TLR9 however, was higher on alveolar macrophages than on monocytes (rMFI, 218.4 ± 187.3 vs. 4.4 ± 1.4, p < 0.05) while the intracellular expression of the receptor and the proportion of TLR9 positive cells were similar in both cell types. TLR2, TLR4 and TLR9 mRNA expression was lower in bronchoalveolar cells than in monocytes.Pam3Cys, LPS, and M.tb DNA upregulated TLR2, TLR4 and TLR9 mRNA in both, bronchoalveolar cells and monocytes. Corresponding with the reduced surface and mRNA expression of TLR2, Pam3Cys induced lower production of TNF-α, IL-1β and IL-6 in bronchoalveolar cells than in monocytes. Despite comparable expression of TLR4 on both cell types, LPS induced higher levels of IL-10 in monocytes than in alveolar macrophages. M.tb DNA, the ligand for TLR9, induced similar levels of cytokines in both cell types.

Conclusion

The TLR expression profile of autologous human alveolar macrophages and monocytes is not identical, therefore perhaps contributing to compartmentalized immune responses in the lungs and systemically. These dissimilarities may have important implications for the design and efficacy evaluation of vaccines with TLR-stimulating adjuvants that target the respiratory tract.     

Effects of Environmental Heterogeneity and Disturbance on the Native and Non-native Flora of Desert Springs

Vegetation often is used as a decision variable for conservation and resource management. Because time and money are limited, it is useful to identify predictable relationships between measures of vegetation diversity or status, the physical environment, and disturbance; native and non-native plants may have different functional responses. Working towards development of effective, practical strategies for management and ecological restoration in the Spring Mountains, an isolated mountain range in the eastern Mojave Desert (Nevada, USA) that is a focus of regional conservation planning, we examined whether native and non-native assemblages of spring-associated perennial plants have predictable relationships with elevation, springbrook length, and various land uses. We also tested whether elevation, springbrook length, and overall disturbance were associated with the degree of predictability of local species presence and absence. Consistent with work in other systems, species richness and cover of native plants tended to decrease as intensity of disturbance increased, whereas species richness (but not cover) of non-native plants tended to peak with intermediate disturbance. Our results may suggest that invasions of non-native plants at springs in the Spring Mountains are relatively recent, and that rapid restoration and management actions may help protect ecological processes and viability of native plants. Ability to predict the order in which individual species are likely to be extirpated from or colonize springs was limited, perhaps reflecting considerable environmental heterogeneity among springs.     

Oxaliplatin enhances TRAIL-induced apoptosis in gastric cancer cells by CBL-regulated death receptor redistribution in lipid rafts

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a member of the tumor necrosis factor family that selectively induces apoptosis in cancer cells. However, gastric cancer cells are insensitive to TRAIL. In the present study, we show that oxaliplatin enhanced TRAIL-induced apoptosis of MGC803, BGC823, and SGC7901 cells. Oxaliplatin promoted death receptor 4 (DR4) and death receptor 5 (DR5) clustering into aggregated lipid rafts, while the cholesterol-sequestering agent nystatin partially prevented lipid raft aggregation, DR4 and DR5 clustering, and reduced apoptosis. Furthermore, the expression of the casitas B-lineage lymphoma (Cbl) family was downregulated by oxaliplatin. Transfection of c-Cbl or Cbl-b partially reversed oxaliplatin-induced lipid raft aggregation. These results indicated that oxaliplatin enhanced TRAIL-induced gastric cancer cell apoptosis at least partially through Cbl-regulated death receptor redistribution in lipid rafts.     

Origin and diversification of the soldier meadow springsnails (Hydrobiidae: Pyrgulopsis), a species flock in the northwestern Great Basin, United States

The large, western North American hydrobiid gastropod genusPyrgulopsis (commonly known as springsnails) includes a seriesof locally endemic faunas that are thought to be species flocks.Although these assemblages are of interest from the evolutionary,biogeographic and conservation perspectives, their monophylyand phylogenetic relationships have yet to be rigorously evaluated.Here we present a molecular phylogeny (based on mitochondrialsequence data) of a putative flock of four thermal spring-dwellingspringsnails that is distributed in many sites in Soldier Meadowand a single locality in Bog Hot Valley (northwestern Nevada).Our analyses support monophyly of this assemblage (‘SoldierMeadow clade’) and a close relationship with other regionalspecies and suggest that the invasion of thermal habitats bythese springsnails occurred independently of other such radiationswithin the genus. The divergence of the Soldier Meadow claderelative to its sister group is substantial (6.79–10.36%for COI, 10.35–15.88% for NDI), suggesting a split inthe early Pliocene, based on the application of a COI clockfor Pyrgulopsis. The splits within the Soldier Meadow cladeinto three main subunits also appear to be old events, basedon their 5.78–8.54% COI divergence relative to each other.These findings are consistent with a long history of springsnailevolution in Soldier Meadow, which is intriguing given thatthis basin was flooded by Lake Lahontan during periods of theearly and middle Pleistocene. We suggest that progenitors ofthe contemporary fauna survived in high elevation springs thatmay have been present in the basin during these pluvial periodsand subsequently colonized contemporary habitats following thetermination of the extreme Lake Lahontan highstands. We speculatethat the broadly disjunct population (of P. militaris) in BogHot Valley, which is consistently nested within the SoldierMeadow clade in our phylogenetic analyses, is either a vicariantrelict of a spring zone that may have once extended betweenthese two areas; or was founded by a past ‘jump’dispersal event from Soldier Meadow. Phylogeographic structureof springsnail populations in Soldier Meadow bears the strongstamp of geologically recent, allopatric diversification, perhapsreflecting the short time that basin floor habitats have beenoccupied. We describe a new species (P. varneri) for a seriesof recently discovered populations that are monophyletic, substantiallydivergent and morphologically distinctive. Additional studieswill be necessary to confidently assess the taxonomic statusof morphologically distinctive P. limaria and P. umbilicata,which are shown herein to be little divergent genetically; anda recently discovered minute springsnail that is morphologicallydivergent yet closely similar genetically to P. notidicola. (Received 20 December 2006; accepted 22 February 2007)     

Predicting susceptibility for SARS‐CoV‐2 infection in domestic and wildlife animals using ACE2 protein sequence homology

The article is presenting a bioinformatics based method predicting susceptibility for SARS‐CoV‐2 infection in domestic and wildlife animals. Recently, there were reports of cats and ferrets, dogs, minks, golden hamster, rhesus monkeys, tigers, and lions testing for SARS‐CoV‐2 RNA which indicated for the possible interspecies viral transmission. Our method successfully predicted the susceptibility of these animals for contracting SARS‐CoV‐2 infection. This method can be used as a screening tool for guiding viral RNA testing for domestic and wildlife animals at risk of getting COVID‐19. We provide a list of the animals at risk of developing COVID‐19 based on the susceptibility score.     

Cyclic peptides. XIX. Synthesis, conformation, and biological activity of cyclo(-Pro-Val-Pro-Val-) with L-L-L-L and L-D-L-D sequences

cyclo(-L-Pro-L-Val-L-Pro-L-Val-) (1L) and cyclo(-L-Pro-D-Val-L-Pro-D-Val-) (1D) were synthesized by the conventional method for peptide synthesis. Conformations of 1L and 1D in solution were studied. Compound 1L has a cis-trans-cis-trans backbone conformation with C2 symmetry in CDCl3. This conformation is slightly different from that in crystalline state and in DMSO-d6 solution. Compound 1D has a cis-trans-cis-trans conformation in DMSO-d6 and an all-trans conformation in trifluoroethanol-d3. Compound 1L retarded stem-growth of rice seedlings and, in contrast, compound 1D promoted root-growth of rice seedlings.     

Associations among spring-dependent aquatic assemblages and environmental and land use gradients in a Mojave Desert mountain range

In many arid landscapes, springs provide the only reliable source of water. Accordingly, both native species and human land uses, including diversion of water, livestock grazing, and recreation, tend to concentrate around springs and spring‐fed riparian areas. We examined whether species richness and composition of aquatic macroinvertebrates at 45 springs in the Spring Mountains, an isolated mountain range in the eastern Mojave Desert (Nevada, USA), could be predicted using readily measured environmental gradients and estimates of disturbance intensity. The Spring Mountains is a focus of regional conservation planning, and managers are charged with prioritizing its springs for conservation and rehabilitation. Our results suggested that species richness of aquatic macroinvertebrates in the Spring Mountains system may be greatest at intermediate levels of natural and human disturbance. Discharge and springbrook length appeared to be only weakly correlated with species richness, whereas neither elevation, nor water temperature, nor electrical conductance was significantly associated with species richness. Nestedness analyses demonstrated that species present in relatively depauperate locations tended to be subsets of the species present in locations that were richer in species, but that pattern did not appear to be driven by either disturbance intensity or by the environmental variables we measured. Disturbance intensity was not associated with the extent to which species presences and absences were predictable. Although our results should not be interpreted to mean that major environmental gradients and disturbance intensity have no effect on distributional patterns of aquatic invertebrates in the Spring Mountains, the ability of these variables to serve as predictors of species richness and composition may be relatively low. Springs and other wetlands in arid landscapes are characterized by isolation and unpredictable disturbances, and faunal responses to environmental gradients may tend to be individualistic and taxon‐specific.