Small Subunit Research Articles

Specific long-term changes in anti-SARS-CoV-2 IgG modifications and antibody functions in mRNA, adenovector, and protein subunit vaccines.

Various vaccine platforms were developed and deployed against the COVID-19 disease. The Fc-mediated functions of IgG antibodies are essential in the adaptive immune response elicited by vaccines. However, the long-term changes of protein subunit vaccines and their combinations with messenger RNA (mRNA) vaccines are unknown. A total of 272 serum and plasma samples were collected from individuals who received first to third doses of the protein subunit Medigen, the mRNA (BNT, Moderna), or the adenovector AstraZeneca vaccines. The IgG subclass level was measured using enzyme-linked immunosorbent assay, and Fc-N glycosylation was measured using liquid chromatography coupled to tandem mass spectrometry. Antibody-dependent-cellular-phagocytosis (ADCP) and complement deposition (ADCD) of anti-spike (S) IgG antibodies were measured by flow cytometry. IgG1 and 3 reached the highest anti-S IgG subclass level. IgG1, 2, and 4 subclass levels significantly increased in mRNA- and Medigen-vaccinated individuals. Fc-glycosylation was stable, except in female BNT vaccinees, who showed increased bisection and decreased galactosylation. Female BNT vaccinees had a higher anti-S IgG titer than that of males. ADCP declined in all groups. ADCD was significantly lower in AstraZeneca-vaccinated individuals. Each vaccine produced specific long-term changes in Fc structure and function. This finding is critical when selecting a vaccine platform or combination to achieve the desired immune response.

A NOVEL SPECIES OF MYXOBOLUS (CNIDARIA: MYXOSPOREA: MYXOBOLIDAE) FROM THE GILL OF GREEN SUNFISH, LEPOMIS CYANELLUS (PERCIFORMES: CENTRARCHIDAE), FROM THE BLACK RIVER OF NORTHEASTERN ARKANSAS.

During March 2023, 7 green sunfish (Lepomis cyanellus) and 2 bluegill (Lepomis macrochirus) were collected from the Black River (White River drainage) in Lawrence County, Arkansas. In addition, during March 2023 and again in May-June 2023, 13 L. cyanellus and 6 L. macrochirus were taken from Butcherknife and Big Fork creeks (Ouachita River drainage), Polk County, Arkansas, 9 L. cyanellus were collected from the Caddo River, Montgomery County, Arkansas, and 5 green sunfish were taken from Clear Creek at Savoy, Washington County, Arkansas. All fish had their gill, gallbladder, fins, integument, musculature, and other major organs examined for myxozoans. The gill of 1 of 34 (3%) L. cyanellus was infected with a new myxozoan, Myxobolus fergusoni n. sp. Qualitative and quantitative morphological data were obtained from fresh myxospores, and molecular data consisted of a 1,933-base-pair sequence of the partial small subunit (SSU) ribosomal RNA (rRNA) gene. Phylogenetic analysis grouped M. fergusoni n. sp. with other centrarchid-infecting myxobolids from North America and placed this cluster in a larger clade comprising myxozoans that infect North American and European esocids, a North American aphredoderid, European percids, and a gasterosteid from Japan. Myxobolus fergusoni n. sp. infects the gill arches of L. cyanellus, similar to Myxobolus cartilaginis (Hoffman, Putz, and Dunbar, 1965), which was described from head cartilage, gill arches, and large fin rays of L. cyanellus. Another is Myxobolus mesentericusKudo, 1920, which was described from the viscera of green sunfish. A large polysporic plasmodium filled with myxospores was present in a basifilamental location associated with multiple gill filaments at their junction with the gill arch. The intact plasmodium replaced connective tissue within the arch but elicited only mild proliferation of overlying epithelium and a minimal host inflammatory response. This is the third time a myxozoan has been described from L. cyanellus, as well as being the first time it has been described from an Arkansas specimen.

First Report of Powdery Mildew on Sophora flavescens Caused by Erysiphe diffusa in China.

Sophora flavescens (Fabaceae) is a deciduous subshrub which has been used in Chinese popular medicine for a long history (He et al. 2015). In June 2023, severe powdery mildew symptoms were observed on wild S. flavescens plants on Longwen hill of Guizhou Normal University, Guiyang, China. The incidence was approximately 80% among 100 S. flavescens plants observed. Almost all leaves were infected. Mycelia occurred on both adaxial and abaxial leaf surfaces, petioles, and stems, forming small-to-large patches. Hyphae were hyaline, 5 to 7 μm wide. Hyphal appressoria were solitary. Conidiophores were erect, straight to somewhat flexuous, and 45 to 120 µm long (n = 50). Foot cells were subcylindrical to slightly curved, followed by 2 to 3 shorter cells. Conidia formed singly, were ovoid to cylindrical, 26 to 42 × 12 to18 µm (n = 50). Based on these morphological characteristics, the powdery mildew fungus was tentatively identified as Erysiphe diffusa (Braun and Cook 2012). To confirm the identification, the ribosomal DNA internal transcribed spacer (ITS) and the ribosomal large subunit (LSU) region were amplified and sequenced using primer pairs ITS1/ITS4 (White et al. 1990) and NL1/NL4 (Ziemiecki et al. 1990), respectively. The obtained 647-bp ITS sequence (GenBank accession no. PP130131) displayed 100% identity with the ITS sequences of E. diffusa. The obtained 618-bp LSU sequence (GenBank accession no. PP693303) displayed 100% identity with the ITS sequences of E. diffusa (MT325922 and MT628019) and E. manihoticola (MT106658 and MT106660). Using a phylogenetic tree based on the combined ITS-LSU data, the isolate was grouped in a clade with the E. diffusa strain (GenBank accession no. LC777871). To fulfill Koch's postulates, leaves of three healthy potted S. flavescens plants were inoculated by gently pressing with diseased leaves. Non-inoculated plants were used as controls. All plants were incubated in a greenhouse at 25 ± 2°C, 80% relative humidity. After 15 days, typical powdery mildew symptoms were observed on the inoculated plants, whereas no symptoms were found on the control plants. The reisolated fungus from the inoculated S. flavescens was morphologically identical to that on naturally diseased plants, and the ITS sequence of the reisolated fungus showed 100% identity with PP130131. As the causal fungus of soybean powdery mildew, E. diffusa is known to infect papaya and other legumes, including Lens culinaris and Mimosa caesalpiniifolia (Attanayake et al. 2009; Luz et al. 2019). Particularly, E. diffusa has been previously reported to infect S. flavescens in the United Kingdom (Jones and Baker 2007; Bradshaw et al. 2023), but this is the first report of S. flavescens powdery mildew caused by E. diffusa in China. This work further expands the geographical range of E. diffusa-infected S. flavescens plants.

Genetic characterization of Cryptosporidium spp. in the North African hedgehog (Atelerix algirus) in the Canary Islands, Spain.

The North African hedgehog (Atelerix algirus) is an introduced species from Northwest Africa and is currently distributed in the Canary Islands. This species of hedgehog has been studied as a reservoir of enteropathogens, including Cryptosporidium spp. However, there are no data at species level. Therefore, the aim of the present study was to identify the Cryptosporidium species present in a population of hedgehogs (n = 36) in the Canary Islands. Molecular screening was performed using conventional polymerase chain reaction (PCR) targeting the small subunit ribosomal RNA (18S rRNA) gene of Cryptosporidium spp. Seven of the 36 fecal samples (19.45%) were positive and confirmed by nested PCR targeting the 18S rRNA gene and Sanger sequencing. Cryptosporidium parvum and Cryptosporidium muris were identified in 11.1% (4/36) and 5.6% (2/36) of the samples, respectively, while one sample could only be identified at the genus level. The zoonotic subtypes IIdA15G1 (n = 1), IIdA16G1b (n = 1), and IIdA22G1 (n = 1) of C. parvum were identified by nested PCR followed by analysis of the 60kDa glycoprotein (gp60) gene sequence. This study is the first genetic characterization of Cryptosporidium spp. in A. algirus, identifying zoonotic species and subtypes of the parasite.

Unveiling Blastocystis epidemiology in Morocco: subtype diversity among clinical patients with and without gastrointestinal manifestations in the Meknes region.

Blastocystis is an intestinal protist frequently identified in humans and other animals, though its clinical significance remains controversial. This study aimed to determine the prevalence and genetic diversity of Blastocystis in faecal samples from symptomatic (n = 55) and asymptomatic (n = 50) individuals seeking medical care in Meknes, Morocco. Detection of the protist was accomplished through coproparasitological examination and culture in Jones medium. Culture-positive samples were subjected to molecular analyses (PCR and Sanger sequencing) based on sequences of the small subunit ribosomal RNA gene. Epidemiological questionnaires on demographics and potential risk factors were collected from participating patients. The overall Blastocystis infection rate was 51.4% (54/105), with no differences between symptomatic (52.7%, 29/55) and asymptomatic (50.0%, 25/50) individuals. Sequence analyses identified three Blastocystis subtypes, with ST3 being the most prevalent (42.0%), followed by ST1 (34.0%), and ST2 (12.0%). Regarding intra-subtype diversity, allele 4 was found within ST1; alleles 11/12 and alleles 34/36 (alone or in combination) were identified within ST2 and ST3 respectively. Allele 34 in ST3 (40.8%) and allele 4 in ST1 (34.7%) were the most common genetic variants circulating in the surveyed clinical population. A statistically significant association between ST2 and the presence of flatulence was observed. This is the first study assessing the epidemiology and genetic diversity of Blastocystis sp. in the Meknes region, Morocco.

Integrative identification of Aphelenchoides fragariae (Nematoda: Aphelenchoididae) parasitizing Fuchsia hybrid in China

The strawberry crimp nematode (Aphelenchoides fragariae) is a serious pathogen of ornamental crops and an important quarantine object in approximately 50 countries and regions including China. One nematode population within the genus Aphelenchoides was discovered from diseased leaves of fuchsia plants (Fuchsia × hybrid Voss.) in Chengdu city, Sichuan province of China. Morphological and morphometric data were obtained using light microscopy and scanning electron microscopy. After detailed examination, the species was identified as A. fragariae. Three rDNA sequences of this species, including partial rRNA small subunit, D2-D3 expansion domains of the rRNA large subunit and internal transcribed spacer, were amplified and sequenced. Bayesian trees inferred from these three rDNA sequences were constructed, revealing that this species is placed in a high support monophyletic clade with A. fragariae but clearly separated from all other Aphelenchoides species. Moreover, host-suitability tests showed that the Aphelenchoides population not only can harm and reproduce in F. hybrid, but also in Fragaria ananassa and Pteris vittata (two common hosts of A. fragariae). In conclusion, the study confirmed A. fragariae identity of the nematode from F. hybrid in Chengdu city based on morphology, molecular analysis and host-suitability tests. To our knowledge, this is the first molecular and morphological confirmation of A. fragariae in China, and F. hybrid was first discovered to be attacked by A. fragariae.

Combined Effect of Basic Antiherpetic Drugs with a New Inhibitor of the Terminase Complex of Herpes Simplex Virus Type 1 in Vero Cell Cultures.

Carriers of herpes simplex virus type 1 (HSV-1) account for more than 90% of the global population. Infection manifests itself in the formation of blisters and ulcers on the face or genitals and can cause blindness, encephalitis, and generalized infection. All first- and second-line modern antiherpetic drugs selectively inhibit viral DNA polymerase. The purine-benzoxazine conjugate LAS-131 ((S)-4-[6-(purin-6-yl)aminohexanoyl]-7,8-difluoro-3,4-dihydro-3-methyl-2H-[1,4]benzoxazine), which we have described earlier, uses the large subunit of the HSV-1 terminase complex as a biotarget and selectively inhibits HSV-1 reproduction in vitro. Basically new results were for the first time obtained to characterize the combined effect on human herpesvirus infection for LAS-131 used in combination with practically significant antiviral compounds, including the nucleoside analogs acyclovir (ACV), penciclovir (PCV), ganciclovir (GCV), brivudine (BVdU), iododeoxyuridine (IdU), and adenine arabinoside (Ara-A); the nucleoside phosphonate analog cidofovir (CDV); and the pyrophosphate analog foscarnet (FOS). A cytopathic effect (CPE) inhibition assay showed that the drug concentration that inhibited the virus-induced CPE by 50% decreased by a factor of 2 (an additive effect, FOS) or more (a synergistic effect; ACV, PCV, GCV, IdU, BVdU, Ara-A, and CDV) when the drugs were used in combination with LAS-131. Nonpermissive conditions for HSV-1 reproduction were thus created at lower drug concentrations, opening up new real possibilities to control human herpesvirus infection.

Isolation, whole genome sequencing and application of a broad-spectrum Salmonella phage.

Salmonella is considered as one of the most common zoonotic /foodborne pathogens in the world. The application of bacteriophages as novel antibacterial agents in food substrates has become an emerging strategy. Bacteriophages have the potential to control Salmonella contamination.We have isolated and characterized a broad-spectrum Salmonella phage, SP154, which can lyse 9 serotypes, including S. Enteritidis, S. Typhimurium, S. Pullorum, S. Arizonae, S. Dublin, S. Cholerasuis, S. Chester, S. 1, 4, [5], 12: i: -, and S. Derby, accounting for 81.9% of 144 isolates. SP154 showed a short latent period (40min) and a high burst size (with the first rapid burst size at 107 PFUs/cell and the second rapid burst size at approximately 40 PFUs/cell). Furthermore, SP154 activity has higher survival rates across various environmental conditions, including pH 4.0-12.0 and temperatures ranging from 4 to 50°C for 60min, making it suitable for diverse food processing and storage applications. Significant reductions in live Salmonella were observed in different foods matrices such as milk and chicken meat, with a decrease of up to 1.9 log10 CFU/mL in milk contamination and a 1 log10 CFU/mL reduction in chicken meat. Whole genome sequencing analysis revealed that SP154 belongs to the genus Ithacavirus, subfamily Humphriesvirinae, within the family Schitoviridae. Phylogenetic analysis based on the terminase large subunit supported this classification, although an alternate tree using the tail spike protein gene suggested affiliation with the genus Kuttervirus, underscoring the limitations of relying on a single gene for phylogenetic inference. Importantly, no virulence or antibiotic resistance genes were detected in SP154. Our research highlights the potential of using SP154 for biocontrol of Salmonella in the food industry.

Population structure and haplotype network analyses of Hyalomma anatolicum based on the large subunit ribosomal RNA (16S rRNA) gene.

Hyalomma anatolicum, an Anatolian hard tick is a well-recognized vector involved in the transmission of various pathogens to animals and humans. The present study elucidated the population structure and haplotype network of H. anatolicum based on the mitochondrial large subunit ribosomal RNA (16S rRNA) gene sequence. The population structure and haplotype network analysis of 75 sequences archived in the GenBank, including the 15 sequences generated herein, yielded 24 haplotypes. Haplotype 1 (Hap_1) was the predominant haplotype consisting of 45 sequences from India, China, Pakistan, Turkey, Egypt, Iraq, and Tajikistan. The complete haplotype network exhibited a stellate conformation, highlighting a recent population expansion. The overall dataset, together with the sequences corresponding to India, China, and Pakistan, showed a high haplotype (0.638 ± 0.065, 0.671 ± 0.103, 0.753 ± 0.099, and 0.854 ± 0.061, respectively) and low nucleotide (0.00407 ± 0.00090, 0.00525 ± 0.00196, 0.00680 ± 0.00233, and 0.00453 ± 0.00056, respectively) diversity, further emphasized a recent population expansion. The neutrality indices including Tajima's D, Fu and Li's D, and Fu and Li's F for the complete dataset (- 2.661, - 6.008, and - 5.649, respectively) as well as for the sequences from India (- 2.223, - 3.414, and - 3.567, respectively) were negative, suggesting deviation from neutrality and a recent population expansion. The present study provided novel insights into the population structure and haplotype networks of H. anatolicum based on the mitochondrial 16S rRNA gene, and the different tests inferred a low genetic differentiation and suggested a recent population expansion of this economically important tick species.

Anti-SARS-CoV-2 Omicron BA.5 RBD antibody titers generated by the protein subunit vaccine Abdala in breast milk

ObjectiveIn this work, we investigate the presence of antibodies against OmicronBA.5 RBD in breast milk from puerperal women, vaccinated during pregnancy with the protein subunit vaccine Abdala, based on the Wuhan-Hu-1 RBD. Materials and methodsBreast milk samples were collected between September 30 and November 25, 2021, from 2 groups of participants: 5- and 9-weeks after the third dose of the vaccine. Antibody titers against OmicronBA.5 RBD and Wuhan-Hu-1 RBD were evaluated simultaneously by a homemade ELISA. The relationship of antibody titers with the parity at the beginning of vaccination and with the participants health status was assessed. ResultsBinding of OmicronBA.5 RBD, such as Wuhan-Hu-1, to IgA and IgG antibodies was detected in all breast milk samples, although the anti-RBD-Wuhan-Hu-1 titers were higher. The immune response to OmicronBA.5 could be related to the glycosylation of the Abdala RBD, expressed in Pichia pastoris, which contributes to the immunogenicity and to the exposure of epitopes that are conserved in pandemic variants, such as Omicron. Antibody titers against both RBDs were lower in participants with chronic diseases and in multiparous. ConclusionsThis is the first report of simultaneous detection of antibody titers, against the Wuhan-Hu-1 RBD and OmicronBA.5 RBD, in breast milk from puerperal women vaccinated with 3 doses a COVID-19 protein subunit vaccine. There is a tendency to decrease the antibody response after vaccination, due to chronic diseases and multiparity.

SARS-CoV-2 multi-epitope subunit vaccine proof-of-concept derived from the insilico study with protein expression in Escherichia coli BL21

The protein subunit vaccine is the most considerably developed SARS-CoV-2 vaccine, according to the WHO vaccine tracker in 2023. The acceleration of vaccine development in 2 years of eradicating the COVID-19 pandemic is attainable due to the role of bioinformatics. This paper aims to evaluate strategies for developing multi-epitope SARS-CoV-2 recombinant vaccines in silico with high protein expression in Escherichia coli vector plasmid. The study was conducted by analysing SARS-CoV-2 epitopes using immunoinformatic tools provided by IEDB, codon optimisation, rare codon analysis, plasmid design, and ribosomal binding site (RBS) analysis were analysed using RNA structure 6.4, gene cloning by E. coli DH5α and protein expression by E. coli BL21. Each epitope peptide candidate was linked to a flexible linker sequence (GGGGS). GelAnalyzer 19.1 was utilised to determine the protein band of SDS-PAGE. The immunoinformatic study obtained a multi-epitope of the recombinant SARS-COV-2 vaccine with 7 epitopes for HLA-I allele candidates and 4 for HLA-II. It is demonstrated that the candidate vaccine protein was successfully cloned in E. coli DH5α and expressed in E. coli BL21. The result of this study will benefit the development of a SARS-CoV-2 protein recombinant vaccine that is safe, affordable, and efficient to induce T-cell response.

An updated LSU database and pipeline for environmental DNA identification of arbuscular mycorrhizal fungi

Recent work established a backbone reference tree and phylogenetic placement pipeline for identification of arbuscular mycorrhizal fungal (AMF) large subunit (LSU) rDNA environmental sequences. Our previously published pipeline allowed any environmental sequence to be identified as putative AMF or within one of the major families. Despite this contribution, difficulties in implementation of the pipeline remain. Here, we present an updated database and pipeline with (1) an expanded backbone tree to include four newly described genera and (2) several changes to improve ease and consistency of implementation. In particular, packages required for the pipeline are now installed as a single folder (conda environment) and the pipeline has been tested across three university computing clusters. This updated backbone tree and pipeline will enable broadened adoption by the community, advancing our understanding of these ubiquitous and ecologically important fungi.

A Novel Inhibitor of Translation Initiation Factor eIF5B in Saccharomyces cerevisiae.

The eukaryotic translation initiation factor eIF5B is a bacterial IF2 ortholog that plays an important role in ribosome joining and stabilization of the initiator tRNA on the AUG start codon during the initiation of translation. We identified the fluorophenyl oxazole derivative 2,2-dibromo-1-(2-(4-fluorophenyl)benzo[d]oxazol-5-yl)ethanone quinolinol as an inhibitor of fungal protein synthesis using an in vitro translation assay in a fungal system. Mutants resistant to this compound were isolated in Saccharomyces cerevisiae and were demonstrated to contain amino acid substitutions in eIF5B that conferred the resistance. These results suggest that eIF5B is a target of potential antifungal compound and that mutation of eIF5B can confer resistance. Subsequent identification of 16 other mutants revealed that primary mutations clustered mainly on domain 2 of eIF5B and secondarily mainly on domain 4. Domain 2 has been implicated in the interaction with the small ribosomal subunit during initiation of translation. The tested translation inhibitor could act by weakening the functional contact between eIF5B and the ribosome complex. This data provides the basis for the development of a new family of antifungals.

Phylogenetic analysis of Ellipsomyxa species (Myxosporea) and description of Ellipsomyxa gordeyi n. sp. from the gall bladder of mullets (Mugiliformes: Mugilidae) in Nha Trang Bay of the East Sea, Vietnam

The coastal waters of Vietnam are home to a wide diversity of fishes, but the parasite diversity of these potential hosts is much less well characterized. To begin addressing this knowledge gap, we carried out surveys of myxozoan parasites in fishes collected from Nha Trang Bay in Vietnam's East Sea in 2018–2019. Mugilid fishes were collected in March–April 2018, January–February 2019, and November–December 2019, and examined for myxozoans. Myxospores consistent with those of the genus Ellipsomyxa were found in the gall bladder of four mullet species, and we thoroughly characterized those from Planiliza melinoptera. Myxospores were elliptoid and devoid of striation, with a distinct sinuous suture line. Polar capsules were pyriform and oriented toward the poles of the spore. Morphological features were compared to nominal species and this species from Vietnam was distinct. Phylogenetic analysis based on partial small subunit rDNA sequence revealed that broadly, Ellipsomyxa species split into three phylogenetic lineages, and although in some branches there are groupings by host family, habitat or locality, there are no clear phylogenetic patterns. The new species we encountered in P. melinoptera had a close sister relationship with Ellipsomyxa adlardi, with both species part of a larger subclade within the Ellipsomyxa lineage. Despite this phylogenetic similarity, these species were morphologically distinct, and partial large subunit DNA sequences were only 93% similar to each other. A combination of the morphological characteristics and molecular data suggest that this is an undescribed species and we propose the name Ellipsomyxa gordeyi n. sp.

Diversity and New Species of Ascomycota from Bamboo in China.

Bamboo is an economically important crop that has gained prominence as an alternative to wood to reduce deforestation and ecosystem destruction. Diseases of bamboo that typically occur on leaves and stems can cause significant loss, reducing the quality and yield of the bamboo. However, there are few reports identifying the fungal species diversity and potential pathogens of bamboo. Here, we describe four new species of plant fungi from the leaves of bamboo within Fujian provinces, China. Fungi were isolated from diseased leaves collected within Fujian province and identified based on their morphological characteristics and multilocus phylogenies using nucleotide sequences derived from combined datasets of the intervening 5.8S nrRNA gene (ITS), the 28S large subunit of nuclear ribosomal RNA gene (LSU), the large subunit of RNA polymerase I (rpb1), the translation elongation factor 1-α gene (tef1-α), and the partial beta-tubulin gene (tub2). These analyses helped reveal and clarify taxonomic relationships in the family Magnaporthaceae. The new species of bambusicolous fungi identified include two species of Bifusisporella, described as B. fujianensis sp. nov. and B. bambooensis sp. nov., and two species of Apiospora, described as A. fujianensis sp. nov. and A. fuzhouensis sp. nov. This study further expands the characterization and distribution of fungi associated with bamboo.

IGF2BP3/NCBP1 complex inhibits renal tubular senescence through regulation of CDK6 mRNA stability

Renal aging and the subsequent rise in kidney-related diseases are attributed to senescence in renal tubular epithelial cells (RTECs). Our study revealed that the abnormal expression of insulin-like growth factor 2 mRNA-binding protein 3 (IGF2BP3), a reader of RNA N6-methyladenosine, is critically involved in cisplatin-induced renal tubular senescence. In cisplatin-induced senescence of RTECs, the promoter activity and transcription of IGF2BP3 is markedly suppressed. It was due to the down regulation of MYC proto-oncogene (MYC), which regulates IGF2BP3 transcription by binding to the putative site at 1852–1863 of the IGF2BP3 promoter. Overexpression of IGF2BP3 ameliorated cisplatin-induced renal tubular senescence in vitro. Mechanistic studies revealed that IGF2BP3 inhibits cellular senescence in RTECs by enhancing cyclin-dependent kinase 6 (CDK6) mRNA stability and increasing its expression. The inhibition effect of IGF2BP3 on tubular senescence is partially reversed by the knockdown of CDK6. Further, IGF2BP3 recruits nuclear cap binding protein subunit 1 (NCBP1) and inhibits CDK6 mRNA decay, by recognizing m6A modification. Specifically, IGF2BP3 recognizes m6A motif "GGACU" at nucleotides 110–114 in the 5′ untranslated region (UTR) field of CDK6 mRNA. The involvement of IGF2BP3/CDK6 in alleviating tubular senescence was confirmed in a cisplatin-induced acute kidney injury (AKI)-to-chronic kidney disease (CKD) model. Clinical data also suggests an age-related decrease in IGF2BP3 and CDK6 levels in renal tissue or serum samples from patients. These findings suggest that IGF2BP3/CDK6 may be a promising target in cisplatin-induced tubular senescence and renal failure.

Investigating Enterocytozoon bieneusi in pigs farmed in Zhejiang Province, China: Occurrence, genotype identification, evolutionary analysis, and zoonotic risk assessment

Enterocytozoon bieneusi is a widespread intracellular fungus that can infect both humans and animals, making it a significant zoonotic threat. In the current study, a total of 208 fecal samples were assayed to investigate the prevalence of E. bieneusi in pigs reared in Zhejiang Province, China. Employing polymerase chain reaction (PCR) amplification techniques specifically designed to target the internal transcribed spacer (ITS) region of the small subunit ribosomal RNA (rRNA) gene, the results revealed that 78 samples (37.5 %) tested positive for the presence of E. bieneusi. A total of 19 different genotypes of E. bieneusi were detected. Nine of these genotypes were already known: EbpC (n = 36), KIN-1 (n = 10), PigEbITS7 (n = 8), EbpA (n = 6), Henan III (n = 3), PigEbITS5 (n = 2), Henan-IV (n = 1), EbpD (n = 1), and TypeIV (n = 1), and 10 were novel: ZJP-I to ZJP-X (one each). The present investigation revealed that all the nine known genotypes identified in pigs here, have also been previously discovered in humans. Additionally, the novel genotypes of E. bieneusi discovered here were all classified as belonging to Group 1. These findings suggest the potential for cross-species transmission between humans and pigs.

The Proteome Profile of Halimeda macroloba under Elevated Temperature: A Case Study from Thailand

An elevated sea temperature is considered a key abiotic stressor causing thermal stress to intertidal macroalgae and influencing their populations. Halimeda macroloba is an important CaCO3 producer that contributes to the carbonate budget in marine ecosystems. The population decline of this intertidal algal species could lead to considerable declines in both regional and global carbonate production. However, the impact of increasing temperature on the molecular mechanisms and protein profile of calcified H. macroloba is unclear and remains to be explored. In this study, H. macroloba was exposed to 30 °C and 35 °C for 7 days. The whole protein was then extracted using 0.5% SDS and digested using trypsin before an analysis using LC-MS. The protein profile of H. macroloba was characterized using the MaxQuant program aligned with the UniProt database. A total of 407 proteins were identified, and 12 proteins were found to be significantly upregulated or downregulated in response to the elevated temperature. Cell division protein, protein kinase domain-containing protein, phospholipid transport protein, and small ribosomal subunit protein were the significant proteins identified in our dataset. The proteins associated with cell division, cellular metabolic processes, localization, oxidoreductase activity, and biosynthetic process pathways were overexpressed with a more than 2-fold change at a high temperature. An interaction map generated using STITCH revealed that the significant protein change altered the other proteins related to abiotic stress, producing energy and inducing calcification. This information could be useful in understanding how H. macroloba responds to an elevated sea temperature.

An injectable subunit vaccine containing Elongation Factor Tu and Heat Shock Protein 70 partially protects American bison from Mycoplasma bovis infection.

Mycoplasma bovis (M. bovis) is the etiologic agent of high mortality epizootics of chronic respiratory disease in American bison (Bison bison). Despite the severity of the disease, no efficacious commercial vaccines have been licensed for the prevention of M. bovis infection in bison. Elongation factor thermal unstable (EFTu) and Heat Shock Protein 70 (Hsp70, DnaK) are highly conserved, constitutively expressed proteins that have previously been shown to provide protection against M. bovis infection in cattle. To assess the suitability of EFTu and Hsp70 as vaccine antigens in bison, the immune response to and protection conferred by an injectable, adjuvanted subunit vaccine comprised of recombinantly expressed EFTu and Hsp70 was evaluated. Vaccinates developed robust antibody and cellular immune responses against both EFTu and Hsp70 antigens. To assess vaccine efficacy, unvaccinated control and vaccinated bison were experimentally challenged with bovine herpes virus-1 (BHV-1) 4 days prior to intranasal infection with M. bovis. Vaccinated bison displayed reductions in joint infection, lung bacterial loads, and lung lesions compared to unvaccinated controls. Together, these results showed that this subunit vaccine reduced clinical disease and bacterial dissemination from the lungs in M. bovis challenged bison and support the further development of protein subunit vaccines against M. bovis for use in bison.

Loss-of-Function Thr347Ala Variant in the G Protein Subunit-α11 Causes Familial Hypocalciuric Hypercalcemia 2 (FHH2).

To date, only four loss-of-function variants in the GNA11 gene encoding the G protein subunit α11 (Gα11) leading to familial hypocalciuric hypercalcemia 2 (FHH2) have been characterized. Gα11 is involved in calcium-sensing receptor (CaSR) signaling, and loss-of-function variants in GNA11 lead to reduced agonist potency at CaSR and an FHH phenotype. We have identified a family with a heterozygous GNA11 Thr347Ala variant and characterized its impact on calcium homeostasis in FHH2 patients and the signaling properties of CaSR through the Gα11-Thr347Ala variant in vitro. The index patient and her family had clinical, biochemical, and genetic analyses performed. The expression levels of Gα11 and the cell-surface expression levels of CaSR in human embryonic kidney 293A Gq/11 knock-out cells (ΔGq/11-HEK293A) co-transfected with CaSR and Gα11 (wild type (WT) or Thr347Ala) were determined, and the functional properties exhibited by calcium at CaSR were characterized in an inositol monophosphate (IP1) accumulation assay. Heterozygous carriers of the GNA11 Thr347Ala variant had mild asymptomatic hypercalcemia, hypocalciuria, and inappropriately high normal parathyroid hormone (PTH) levels considering their elevated serum calcium levels. Whereas the variant did not impact Gα11 expression or CaSR cell surface expression levels, calcium displayed a moderately but significantly lower agonist potency at CaSR/Gα11-Thr347Ala-transfected cells compared with CaSR/Gα11-WT-transfected cells in the IP1 accumulation assay (EC50 values of 5.67 mM and 4.38 mM, respectively). This identification of a novel GNA11 variant causing FHH2 substantiates the important role of Gα11 for CaSR signaling and Ca2+ homeostasis.