Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Effect of fear of hypoglycaemia on sleep quality of patients with type 2 mellitus diabetes: The mediating role of alexithymia

Background: Patients with type 2 diabetes mellitus (T2DM) commonly experience poor sleep quality. This study aimed to investigate whether alexithymia mediates the association between fear of hypoglycaemia (FoH) and sleep quality in patients with T2DM. Methods: From September 2021 to November 2021, a cross-sectional survey was conducted on 407 patients with T2DM in China. Data collection was made possible through the administration of the Chinese Version of the Worry Scale, Toronto Alexithymia Scale and Chinese version of the Pittsburgh Sleep Quality Index (CPSQI). Multiple linear regression analyses were also performed. Results: A total of 65.6% of the participants were male, and 75.7% were aged 18–40 years. FoH showed a moderate and positive correlation with CPSQI scores (r = 0.308, p < 0.001). Alexithymia was weakly and positively correlated with CPSQI scores (r = 0.185, p < 0.001). Meanwhile, FoH exhibited a moderate and positive correlation with alexithymia (r = 0.422, p < 0.001), and difficulty in identifying (r = 0.414, p < 0.001) and describing feelings (r = 0.416, p < 0.001) and a weak and positive correlation with externally oriented thinking (r = 0.221, p < 0.001). The total effect (β = 0.408, p < 0.001) of FoH on CPSQI comprised not only the direct (β = 0.293, 95% confidence interval: 0.174–0.411, p < 0.001) but also the indirect effect (β = 0.115, p < 0.001) of alexithymia. Conclusions: Alexithymia can mediate the association between FoH and sleep quality. Clinicians should recognize the potential effect of alexithymia and incorporate it in intervention planning and care. Addressing the affective disturbances arising from FoH can enhance emotional expression and sleep quality among T2DM patients

Sleep duration trajectories and all-cause mortality among Chinese elderly: A community-based cohort study

Abstract Background China is among the largest and fastest aging countries. The elderly population is more vulnerable, with higher proportion of inappropriate sleep duration and risk of mortality, compared with young and middle-aged adults. Single-measured sleep duration has been associated with mortality, but the health effects of long-term sleep duration trajectories remain unknown. This study aimed to explore the prospective associations between sleep duration trajectories and all-cause mortality among Chinese elderly. Methods Participants (n = 3,895; median age: 82 years; females: 53.3%) who reported sleep duration in all three surveys (2005, 2008, and 2011) from the community-based Chinese Longitudinal Healthy Longevity Survey (CLHLS) were followed up until 2019 (about 8 years). We identified sleep duration trajectories by latent class mixed model and explored their association with all-cause mortality using Cox hazard proportional regression and Laplace regression models. Further, stratified analysis by demographic characteristics and lifestyles and sensitivity analysis by lag effect, health-related factors, and inverse probability weighting were used to verify the robustness of the association. In addition, we explored the threshold effect of baseline sleep duration on the risk of all-cause mortality. Results We documented 1,881 all-cause deaths during 16,689 person-years of follow-up. Five sleep duration trajectories were identified: moderately increased trajectory (28.1%), rapidly increased trajectory (7.2%), persistent sleep trajectory of 7 h (33.7%), moderately decreased trajectory (21.3%), and rapidly decreased trajectory (9.7%). Compared with the persistent sleep trajectory of 7 h, the multivariable-adjusted HRs (95%CI) for moderately increased trajectory, rapidly increased trajectory, moderately decreased trajectory, and rapidly decreased trajectory were 1.21 (1.08, 1.36), 1.21 (1.01, 1.44), 0.95 (0.82, 1.10), and 0.93 (0.78, 1.11), respectively; and the corresponding difference in median survival time (95%CI) were -0.53 (-1.01, -0.05), -0.43 (0.16, -1.02), 0.26 (-0.34, 0.86), and 0.25 (-0.51, 1.02), respectively. Stratified and sensitivity analyses showed consistent results. Threshold analysis indicated a sharply increased risk of mortality in participants whose sleep exceeds 9 h (HR = 1.20, 95%CI: 1.11, 1.30). Conclusion Compared with the persistent sleep trajectory of 7 h, moderately and rapidly increased sleep duration trajectories were associated with higher subsequent mortality in Chinese elderly. Those who report sleep exceeding 9 h may be at high risk for all-cause mortality

Andrographolide Ameliorates Liver Fibrosis in Mice: Involvement of TLR4/NF-κB and TGF-β1/Smad2 Signaling Pathways

Liver fibrosis is characterized by activated hepatic stellate cells (HSC) and extracellular matrix accumulation. Blocking the activation of HSC and the inflammation response are two major effective therapeutic strategies for liver fibrosis. In addition to the long history of using andrographolide (Andro) for inflammatory disorders, we aimed at elucidating the pharmacological effects and potential mechanism of Andro on liver fibrosis. In this study, liver fibrosis was induced by carbon tetrachloride (CCl4) and the mice were intraperitoneally injected with Andro for 6 weeks. HSC cell line (LX-2) and primary HSC were also treated with Andro in vitro. Treatment of CCl4-induced mice with Andro decreased the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), Sirius red staining as well as the expression of α smooth muscle actin (α-SMA) and transforming growth factor- (TGF-) β1. Furthermore, the expression of Toll-like receptor (TLR)4 and NF-κB p50 was also inhibited by Andro. Additionally, in vitro data confirmed that Andro treatment not only attenuated the expression of profibrotic and proinflammatory factors but also blocked the TGF-β1/Smad2 and TLR4/NF-κB p50 pathways. These results demonstrate that Andro prevents liver inflammation and fibrosis, which is in correlation with the inhibition of the TGF-β1/Smad2 and TLR4/NF-κB p50 pathways, highlighting Andro as a potential therapeutic strategy for liver fibrosis

Multilocus genotyping of Giardia duodenalis in captive non-human primates in Sichuan and Guizhou provinces, Southwestern China.

Giardia duodenalis is a common human and animal pathogen. It has been increasingly reported in wild and captive non-human primates (NHPs) in recent years. However, multilocus genotyping information for G. duodenalis infecting NHPs in southwestern China is limited. In the present study, the prevalence and multilocus genotypes (MLGs) of G. duodenalis in captive NHPs in southwestern China were determined. We examined 207 fecal samples from NHPs in Sichuan and Guizhou provinces, and 16 specimens were positive for G. duodenalis. The overall infection rate was 7.7%, and only assemblage B was identified. G. duodenalis was detect positive in northern white-cheeked gibbon (14/36, 38.9%), crab-eating macaque (1/60, 1.7%) and rhesus macaques (1/101, 0.9%). Multilocus sequence typing based on beta-giardin (bg), triose phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) revealed nine different assemblage B MLGs (five known genotypes and four novel genotypes). Based on a phylogenetic analysis, one potentially zoonotic genotype of MLG SW7 was identified in a northern white-cheeked gibbon. A high degree of genetic diversity within assemblage B was observed in captive northern white-cheeked gibbons in Southwestern China, including a potentially zoonotic genotype, MLG SW7. To the best of our knowledge, this is the first report using a MLGs approach to identify G. duodenalis in captive NHPs in Southwestern China

Correction: Multilocus genotyping of Giardia duodenalis in captive non-human primates in Sichuan and Guizhou provinces, Southwestern China.

[This corrects the article DOI: 10.1371/journal.pone.0184913.]

Occurrence and genotyping of Giardia duodenalis and Cryptosporidium in pre-weaned dairy calves in central Sichuan province, China

Giardia duodenalis and Cryptosporidium spp. are common human and animal pathogens. They have increasingly been reported in dairy calves in recent years; however, multilocus genotyping information for G. duodenalis and Cryptosporidium infecting pre-weaned dairy calves in southwestern China is limited. In the present study, the prevalence of G. duodenalis and Cryptosporidium spp. in pre-weaned dairy calves in central Sichuan province was determined and the pathogens were analyzed molecularly. Of 278 fecal samples from pre-weaned dairy calves, 26 (9.4%) were positive for G. duodenalis and 40 (14.4%) were positive for Cryptosporidium spp. Cryptosporidium bovis (n = 28), Cryptosporidium ryanae (n = 5) and Cryptosporidium parvum (n = 7) were detected. All seven C. parvum isolates were successfully subtyped based on the gp60 gene sequence, and only IIdA15G1 was detected. Multilocus sequence typing of G. duodenalis based on beta-giardin (bg), triose phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) genes revealed 19 different assemblage E multilocus genotypes (two known and 17 unpublished genotypes). Based on eBURST analysis, a high degree of genetic diversity within assemblage E was observed in pre-weaned dairy calves in Sichuan province. To the best of our knowledge, this is the first study using multilocus sequence typing and eBURST analysis to characterize G. duodenalis in pre-weaned dairy calves in southwestern China

Occurrence and genotyping of

Giardia duodenalis and Cryptosporidium spp. are common human and animal pathogens. They have increasingly been reported in dairy calves in recent years; however, multilocus genotyping information for G. duodenalis and Cryptosporidium infecting pre-weaned dairy calves in southwestern China is limited. In the present study, the prevalence of G. duodenalis and Cryptosporidium spp. in pre-weaned dairy calves in central Sichuan province was determined and the pathogens were analyzed molecularly. Of 278 fecal samples from pre-weaned dairy calves, 26 (9.4%) were positive for G. duodenalis and 40 (14.4%) were positive for Cryptosporidium spp. Cryptosporidium bovis (n = 28), Cryptosporidium ryanae (n = 5) and Cryptosporidium parvum (n = 7) were detected. All seven C. parvum isolates were successfully subtyped based on the gp60 gene sequence, and only IIdA15G1 was detected. Multilocus sequence typing of G. duodenalis based on beta-giardin (bg), triose phosphate isomerase (tpi) and glutamate dehydrogenase (gdh) genes revealed 19 different assemblage E multilocus genotypes (two known and 17 unpublished genotypes). Based on eBURST analysis, a high degree of genetic diversity within assemblage E was observed in pre-weaned dairy calves in Sichuan province. To the best of our knowledge, this is the first study using multilocus sequence typing and eBURST analysis to characterize G. duodenalis in pre-weaned dairy calves in southwestern China

Characterization of 16 specimens based on multi-locus sequences of <i>bg</i>, <i>tpi</i> and <i>gdh</i> genes.

<p>Characterization of 16 specimens based on multi-locus sequences of <i>bg</i>, <i>tpi</i> and <i>gdh</i> genes.</p