Comparative Metabolite Extraction Protocols from Breast Cancer Mouse Lung Tissue for LC-MS/MS Analysis

Triple-negative breast cancer (TNBC) stands out for its heightened invasiveness, leading to distant metastasis in nearly 46% of cases, with common targets being the brain, lungs, and bones. This subtype is associated with significantly shorter median survival compared to other breast cancer types. Analyzing metabolic compounds in lung tissues affected by breast cancer metastasis provides valuable insights into biological information and regulatory processes. Despite the recognized severity of TNBC spreading to other sites, there are limited reported studies investigating metabolome information in distant organ tissues, particularly the lungs. Therefore, accurately quantifying the abundance of metabolites requires careful extraction procedures. This study aims to investigate and compare extraction protocols for lung tissue metabolites in TNBC mice using liquid chromatography with tandem mass spectrometry (LC-MS/MS). Left lung tissues were collected from mice xenografted with breast cancer. Three different extraction methods were evaluated to assess their metabolite coverage and biochemical compound classes. Our findings revealed distinct differences in metabolite compositions among the three methods. The extraction solvent comprising isopropanol, acetonitrile, and water in a 3:2:2 ratio proved most suitable for studying breast cancer metastasis to lung tissues. This extraction solvent could serve as a protocol for future studies analyzing the lung cancer metabolome in mice

Complete coding sequence characterization and comparative analysis of the putative novel human rhinovirus (HRV) species C and B

<p>Abstract</p> <p>Background</p> <p>Human Rhinoviruses (HRVs) are well recognized viral pathogens associated with acute respiratory tract illnesses (RTIs) abundant worldwide. Although recent studies have phylogenetically identified the new HRV species (HRV-C), data on molecular epidemiology, genetic diversity, and clinical manifestation have been limited.</p> <p>Result</p> <p>To gain new insight into HRV genetic diversity, we determined the complete coding sequences of putative new members of HRV species C (HRV-CU072 with 1% prevalence) and HRV-B (HRV-CU211) identified from clinical specimens collected from pediatric patients diagnosed with a symptom of acute lower RTI. Complete coding sequence and phylogenetic analysis revealed that the HRV-CU072 strain shared a recent common ancestor with most closely related Chinese strain (N4). Comparative analysis at the protein level showed that HRV-CU072 might accumulate substitutional mutations in structural proteins, as well as nonstructural proteins 3C and 3 D. Comparative analysis of all available HRVs and HEVs indicated that HRV-C contains a relatively high G+C content and is more closely related to HEV-D. This might be correlated to their replication and capability to adapt to the high temperature environment of the human lower respiratory tract. We herein report an infrequently occurring intra-species recombination event in HRV-B species (HRV-CU211) with a crossing over having taken place at the boundary of VP2 and VP3 genes. Moreover, we observed phylogenetic compatibility in all HRV species and suggest that dynamic mechanisms for HRV evolution seem to be related to recombination events. These findings indicated that the elementary units shaping the genetic diversity of HRV-C could be found in the nonstructural 2A and 3D genes.</p> <p>Conclusion</p> <p>This study provides information for understanding HRV genetic diversity and insight into the role of selection pressure and recombination mechanisms influencing HRV evolution.</p

First report of blaOXA-181-carrying IncX3 plasmids in multidrug-resistant Enterobacter hormaechei and Serratia nevei recovered from canine and feline opportunistic infections.

Whole-genome sequence analysis of six Enterobacter hormaechei and two Serratia nevei strains, using a hybrid assembly of Illumina and Oxford Nanopore Technologies sequencing, revealed the presence of the epidemic blaOXA-181-carrying IncX3 plasmids co-harboring qnrS1 and ∆ere(A) genes, as well as multiple multidrug resistance (MDR) plasmids disseminating in all strains, originated from dogs and cats in Thailand. The subspecies and sequence types (ST) of the E. hormaechei strains recovered from canine and feline opportunistic infections included E. hormaechei subsp. xiangfangensis ST171 (n = 3), ST121 (n = 1), and ST182 (n = 1), as well as E. hormaechei subsp. steigerwaltii ST65 (n = 1). Five of the six E. hormaechei strains harbored an identical 51,479-bp blaOXA-181-carrying IncX3 plasmid. However, the blaOXA-181 plasmid (pCUVET22-969.1) of the E. hormaechei strain CUVET22-969 presented a variation due to the insertion of ISKpn74 and ISSbo1 into the virB region. Additionally, the blaOXA-181 plasmids of S. nevei strains were nearly identical to the others at the nucleotide level, with ISEcl1 inserted upstream of the qnrS1 gene. The E. hormaechei and S. nevei lineages from canine and feline origins might acquire the epidemic blaOXA-181-carrying IncX3 and MDR plasmids, which are shared among Enterobacterales, contributing to the development of resistance. These findings suggest the spillover of significant OXA-181-encoding plasmids to these bacteria, causing severe opportunistic infections in dogs and cats in Thailand. Surveillance and effective hygienic practice, especially in hospitalized animals and veterinary hospitals, should be urgently implemented to prevent the spread of these plasmids in healthcare settings and communities.IMPORTANCEblaOXA-181 is a significant carbapenemase-encoding gene, usually associated with an epidemic IncX3 plasmid found in Enterobacterales worldwide. In this article, we revealed six carbapenemase-producing (CP) Enterobacter hormaechei and two CP Serratia nevei strains harboring blaOXA-181-carrying IncX3 and multidrug resistance plasmids recovered from dogs and cats in Thailand. The carriage of these plasmids can promote extensively drug-resistant properties, limiting antimicrobial treatment options in veterinary medicine. Since E. hormaechei and S. nevei harboring blaOXA-181-carrying IncX3 plasmids have not been previously reported in dogs and cats, our findings provide the first evidence of dissemination of the epidemic plasmids in these bacterial species isolated from animal origins. Pets in communities can serve as reservoirs of significant antimicrobial resistance determinants. This situation places a burden on antimicrobial treatment in small animal practice and poses a public health threat

Serological Analysis of Human Pandemic Influenza (H1N1) in Thailand

The study was aimed at determining the prevalence of pandemic influenza (H1N1) 2009 among patients with respiratory tract diseases during July-December 2009 using real-time reverse transcription polymerase chain reaction. Haemagglutination inhibition (HI) assay was performed to detect antibody titres against pandemic influenza in 255 medical personnel, 307 members of the general population during the second week of December 2009 in Khon Kaen province, Thailand, and in 100 stored sera collected from people of different age-groups during 2008. The results showed that the pandemic (H1N1) 2009 had occurred during July-December 2009. The results of the HI test after the wave of this outbreak showed that 123 (48%) of the 255 sera collected from the medical personnel, 109 (36%) of the 307 sera obtained from the general population, and only two of the 100 stored sera from 2008 contained antibodies (HI titres ≥40) against pandemic influenza. Antibody against the pandemic (H1N1) 2009 was found in at least one-third of the population. In conclusion, the prevalence of virus and serological data obtained from the study can be used as the serological background level of the Thai population after the July-December pandemic. Finally, the serological data might be useful for outbreak-prevention and control strategies and for the management of vaccination for the pandemic (H1N1) 2009 in Thailand

Different Exercise Intensity Associates with Varied Disease Biomarkers of Guts-Microbiome Genera Change in Rats: Preliminary Study

The microbiome, a community of microorganisms in the body, is currently used as a biomarker in many disease prognoses. Prevotella, Turicibacte, Bacteroides, Firmicutes/Bacteroidetes are frequently used as a biomarker for rheumatoid arthritis, colorectal cancer, and obesity in ordered. The amount of gut microbiota can be changed depending on various factors such as diet, lifestyle, and exercise. However, there is unclear on how the exercise is really effective to be a disease prevention. The present study aims to investigate the different exercise intensities on gut microbiome abundance changes that could be used as a disease biomarker. Eighteen Sprague-Dawley rats were arranged (n=6 per group) into 3 exercise intensity levels on treadmills including non-exercise group, high -exercise group (20 – 25 m/min for 60 min), and light-exercise group (10 – 15 m/min for 60 min). Rats were weighted every 2 days and stools were collected and preserved in DNA/RNA shield each week. The bacterial 16S rDNA of microbiome in feces samples was sequenced and analyzed. After week eighth of the interventions, from operational taxonomic unit (OTUs) abundance, we found that the relative abundance in bacterial genera in Prevotella and Firmicutes/Bacteroidetes were significantly correlated with the experiment timepoints in different exercise intensities (Pearson’s correlation, P<0.05) compare to other genera. The exercise intensities and exercise durations can affect the relative abundance in the bacteria genus which the abundance genus Prevotella and Firmicutes/Bacteroidetes could be used as a new standard biomarker in exercise as a disease prevention and exercise prescriptions. From the funding limitations, we could conclude the research results based on our data and statistic. Future research should utilize a longer investigation period

Avian Influenza H5N1 in Naturally Infected Domestic Cat

We report H5N1 virus infection in a domestic cat infected by eating a pigeon carcass. The virus isolated from the pigeon and the cat showed the same cluster as the viruses obtained during the outbreak in Thailand. Since cats are common house pets, concern regarding disease transmission to humans exists