Purification and Activity Evaluation of a Novel Thrombopoietin Mimetic Peptide.

The emergence of thrombopoietin mimetic peptides presents a promising therapeutic strategy for addressing thrombocytopenia. This particular study aimed to establish a direct, expeditious, and efficient method for modifying and purifying a novel thrombopoietin mimetic peptide. Precursor proteins were subjected to modification utilizing three distinct fatty acids: C25H42O7N2, C39H66O15N4, and C41H70O15N4. Liquid chromatography analyses demonstrated that C41H70O15N4 yielded the most effective modification results. Mass spectrometry findings validated the correspondence between the theoretical and actual molecular weights of each sample. In vivo experiments conducted on normal mice showcased that the C41H70O15N4 modification group exhibited the highest platelet count, peaking at an impressive 5047 × 109/L. This count was approximately twice that of the peak platelet count observed in the dTMP group and four times higher than the control group. Pharmacokinetic investigations revealed that the C41H70O15N4 modification group displayed the lengthiest half-life among beagles, persisting for 128.5 h. This duration was approximately 28.5 times longer than that of the unmodified dTMP group. These findings underscore the effectiveness of the established C41H70O15N4 modification and purification method in preserving the biological activity of the thrombopoietin mimetic peptide. The novel thrombopoietin mimetic peptide showcased notable attributes of simplicity and cost-effectiveness, while also exhibiting a significant platelet-promoting effect and an extended half-life. Consequently, this novel peptide holds substantial significance for advancing the treatment of thrombocytopenia.

Application of a Faecalibacterium 16S rDNA genetic marker for species identification of dog fecal waste.

A dog-associated 16S rDNA genetic marker (ED-1) was designed to detect dog fecal contamination in water through a comparative bioinformatics analysis of Faecalibacterium sequences. For the dog fecal samples, ED-1 had 100% specificity, a high positive rate (89% in dog feces and 92.3% in dog fecal-contaminated water samples), and a low detection limit (107 copies/100 mL) in dog-contaminated water samples. Detection of water samples from seven provinces or cities of China showed that ED-1 was stable enough to be applied in practice. Furthermore, the abundance and diversity of dog gut microbiota from two private house pets (PHP) and Third Military Medical University (TMMU) dogs were estimated by using operational taxonomic units, and the significant differences of dog feces were found, as the PHP dogs have a more diverse diet and closer contact with human than dogs in TMMU. However, ED-1 could detect the feces from the two regions, indicating that ED-1 has good reliability.

Potential application of an Aspergillus strain in a pilot biofilter for benzene biodegradation.

A biofilter with fungus was developed for efficient degradation of benzene, which can overcome the potential risk of leakage commonly found in such services. Results indicated that the optimum parameter values were temperature 40 °C, pH 6, and 500 mg L-1 of the initial benzene concentration. Besides, the empty bed residence time and inlet load range of biofilter were set to 20 s and 21.23-169.84 g m-3 h-1 respectively. Under these conditions, this biofilter can obtain the maximum removal efficiency of more than 90%, the eliminating capacity could be up to 151.67 g m-3 h-1. Furthermore, scanning electron microscopy was used to investigate three filler materials for packing fungus biofilm. This is the first study introducing an Aspergillus strain for benzene removal and these results highlight that the development of this biofilter has the potential scaling-up application as gas-processing of industrial wastes.

Evaluation of Faecalibacterium 16S rDNA genetic markers for accurate identification of swine faecal waste by quantitative PCR.

A genetic marker within the 16S rRNA gene of Faecalibacterium was identified for use in a quantitative PCR (qPCR) assay to detect swine faecal contamination in water. A total of 146,038 bacterial sequences were obtained using 454 pyrosequencing. By comparative bioinformatics analysis of Faecalibacterium sequences with those of numerous swine and other animal species, swine-specific Faecalibacterium 16S rRNA gene sequences were identified and Polymerase Chain Okabe (PCR) primer sets designed and tested against faecal DNA samples from swine and non-swine sources. Two PCR primer sets, PFB-1 and PFB-2, showed the highest specificity to swine faecal waste and had no cross-reaction with other animal samples. PFB-1 and PFB-2 amplified 16S rRNA gene sequences from 50 samples of swine with positive ratios of 86 and 90%, respectively. We compared swine-specific Faecalibacterium qPCR assays for the purpose of quantifying the newly identified markers. The quantification limits (LOQs) of PFB-1 and PFB-2 markers in environmental water were 6.5 and 2.9 copies per 100 ml, respectively. Of the swine-associated assays tested, PFB-2 was more sensitive in detecting the swine faecal waste and quantifying the microbial load. Furthermore, the microbial abundance and diversity of the microbiomes of swine and other animal faeces were estimated using operational taxonomic units (OTUs). The species specificity was demonstrated for the microbial populations present in various animal faeces.

Application of Faecalibacterium 16S rDNA genetic marker for accurate identification of duck faeces.

The aim of this study was to judge the legal duty of pollution liabilities by assessing a duck faeces-specific marker, which can exclude distractions of residual bacteria from earlier contamination accidents. With the gene sequencing technology and bioinformatics method, we completed the comparative analysis of Faecalibacterium sequences, which were associated with ducks and other animal species, and found the sequences unique to duck faeces. Polymerase chain reaction (PCR) and agarose gel electrophoresis techniques were used to verify the reliability of both human and duck faeces-specific primers. The duck faeces-specific primers generated an amplicon of 141 bp from 43.3 % of duck faecal samples, 0 % of control samples and 100 % of sewage wastewater samples that contained duck faeces. We present here the initial evidence of Faecalibacterium-based applicability as human faeces-specificity in China. Meanwhile, this study represents the initial report of a Faecalibacterium marker for duck faeces and suggests an independent or supplementary environmental biotechnology of microbial source tracking (MST).

Using an intervening sequence of Faecalibacterium 16S rDNA to identify poultry feces.

This study was designed to identify poultry feces-specific marker(s) within sequences of Faecalibacterium 16S rDNA for detecting poultry fecal pollution in water. Bioinformatics tools were used in the comparative analysis of 7,458 sequences of Faecalibacterium 16S rDNA, reportedly associated with various poultry (chicken and turkey) and animal species. One intervening sequence (IVS) within between the hypervariable region 1 and the conserved region 2, designated as IVS-p, was found to be unique to poultry feces. Based on this sequence, a PCR assay (PCR-p) was developed. The PCR-p produced an amplicon of 132 bp only in the test when fecal or wastewater samples from poultry were used, but not when using fecal or wastewater samples from other sources. The non-poultry sources included feces of beef or dairy cattle, dog, horse, human, domestic or wild geese, seagull, sheep, swine, and wild turkey. These data indicate that IVS-p may prove to be a useful genetic marker for the specific identification of poultry fecal pollution in environmental waterways. Furthermore, results of data mining and PCR assay indicate that the IVS-p may have a broad geographic distribution. This report represents initial evidence of the potential utility of ribosomal intervening sequences as genetic markers for tracking host sources of fecal pollution in waterways.