Retinitis pigmentosa (RP) is the most common type of inherited retinopathy. At least 69 genes for RP have been identified. A significant proportion of RP, however, remains genetically unsolved. In this study, the genetic basis of a Chinese consanguineous family with presumed autosomal recessive retinitis pigmentosa (arRP) was investigated.
Overall ophthalmic examinations, including funduscopy, decimal best-corrected visual acuity, axial length and electroretinography (ERG) were performed for the family. Genomic DNA from peripheral blood of the proband was subjected to whole exome sequencing. In silico predictions, structural modelling, and minigene assays were conducted to evaluate the pathogenicity of the variant.
A novel homozygous variant (NM_003320.4: c.1379A > G) in the TUB gene was identified as a candidate pathogenic variant in this parental consanguineous pedigree. This variant co-segregated with the disease in this pedigree and was absent in 118 ethnically matched healthy controls. It\'s an extremely rare variant that is neither deposited in population databases (1000 Genomes, ExAC, GnomAD, or Exome Variant Server) nor reported in the literature. Phylogenetic analysis indicated that the Asn residue at codon 460 of TUB is highly conserved across diverse species from tropicalis to humans. It was also completely conserved among the TUB, TULP1, TULP2, and TULP3 family proteins. Multiple bioinformatic algorithms predicted that this variant was deleterious.
A novel missense variant in TUB was identified, which was probably the pathogenic basis for arRP in this consanguineous family. This is the first report of a homozygous missense variant in TUB for RP.

Although tumor-infiltrating lymphocytes (TILs) maintain their ability to proliferate, persist, and eradicate tumors, they are frequently dysfunctional in situ. By performing both whole-genome CRISPR and metabolic inhibitor screens, we identify that nicotinamide phosphoribosyltransferase (NAMPT) is required for T cell activation. NAMPT is low in TILs, and its expression is controlled by the transcriptional factor Tubby (TUB), whose activity depends on the T cell receptor-phospholipase C gamma (TCR-PLCγ) signaling axis. The intracellular level of NAD+, whose synthesis is dependent on the NAMPT-mediated salvage pathway, is also decreased in TILs. Liquid chromatography-mass spectrometry (LC-MS) and isotopic labeling studies confirm that NAD+ depletion led to suppressed glycolysis, disrupted mitochondrial function, and dampened ATP synthesis. Excitingly, both adoptive CAR-T and anti-PD1 immune checkpoint blockade mouse models demonstrate that NAD+ supplementation enhanced the tumor-killing efficacy of T cells. Collectively, this study reveals that an impaired TCR-TUB-NAMPT-NAD+ axis leads to T cell dysfunction in the tumor microenvironment, and an over-the-counter nutrient supplement of NAD+ could boost T-cell-based immunotherapy.

OBJECTIVE: Dysfunction of major cells constituting the aortic wall is the pathological basis for AD development. Determining whether non-coding RNAs can influence AD progression by regulating these cellular functions and identifying some specific non-coding RNAs is of great significance in uncovering molecular mechanisms of the development of AD.
METHODS: Microarray analyses and hierarchical clustering analysis were used to select candidate lncRNAs and miRNAs associated with AD. Dual-luciferase reporter assay, RNA immunoprecipitation, and RNA pull-down assay were performed to verify the direct bonding relationship between genes. The regulatory effects of genes on cell function were examined in a series of experiments.
RESULTS: We found that lnc-OIP5-AS1 was upregulated, whereas miR-143-3p was downregulated in cells treated with angiotensin II (AngII) and AD tissues. Lnc-OIP5-AS1 functioned as a competing endogenous RNA (ceRNA) of miR-143-3p to suppress the proliferation and mobility, but promote apoptosis of HAECs and HASMCs, and simultaneously result in the imbalances between MMP-2/9 and TIMP-2/1 in HASMCs and the excessive secretion of IL-6, IL-1β, and IL-17A of HAAFs. Moreover, overexpression or silence of TUB, a target gene of miR-143-3p, counteracted the influence of miR-143-3p or lnc-OIP5-AS1 on cells, respectively.
CONCLUSIONS: Our findings revealed that lncRNA OIP5-AS1 exacerbates aorta intima, media, and adventitia injury in the development of AD through upregulating TUB via sponging miR-143-3p and also support more detailed future studies by providing a novel molecular basis underlying AD formation.

Photoreceptor disc component (PRCD) is a small protein which is exclusively localized to photoreceptor outer segments, and is involved in the formation of photoreceptor outer segment discs. Mutations in PRCD are associated with retinal degeneration in humans, mice, and dogs. The purpose of this work was to identify PRCD-binding proteins in the retina. PRCD protein-protein interactions were identified when implementing the Ras recruitment system (RRS), a cytoplasmic-based yeast two-hybrid system, on a bovine retina cDNA library. An interaction between PRCD and tubby-like protein 1 (TULP1) was identified. Co-immunoprecipitation in transfected mammalian cells confirmed that PRCD interacts with TULP1, as well as with its homolog, TUB. These interactions were mediated by TULP1 and TUB highly conserved C-terminal tubby domain. PRCD localization was altered in the retinas of TULP1- and TUB-deficient mice. These results show that TULP1 and TUB, which are involved in the vesicular trafficking of several photoreceptor proteins from the inner segment to the outer segment, are also required for PRCD exclusive localization to photoreceptor outer segment discs.

Pseudomonas aeruginosa folliculitis is an infection of the skin commonly associated with swimming pool and hot tub use. It often presents as outbreaks affecting multiple individuals using the same contaminated public water facility. We present a case report of a 50-year-old woman who developed pseudomonal folliculitis after using a hot tub with multiple family members. No other family member developed folliculitis. Factors contributing to susceptibility to P. aeruginosa infection are reviewed.

The aim of this study was to investigate the molecular mechanism of pancreatic islet-derived mesenchymal stem cell (PID-MSC) differentiation into beta-cells in the presence of insulin and leptin resistance stimulators. We determined that beta-cell differentiation was stimulated by glucose, insulin, and leptin. Co-administration of insulin and leptin resulted in greater, at a further stage of differentiation but non-functional beta-cell formation. The levels of p-AKT(Ser473) did not change; SOCS3, PTP1B, p-IRS1(Ser307), PTEN levels increased and p-IRS1(Try) levels decreased due to insulin and leptin co-administration. These findings suggest that co-administration of insulin and leptin to PID-MSCs results in the development of both insulin and leptin resistance together. We showed that this differentiation signaling is mainly mediated by AKT/GSK-3β/β-catenin and Tub. Moreover, β-catenin and Tub were linked to each other in the nucleus under this condition. Furthermore, we found that Tub and β-catenin contributes to insulin production by increasing the expression of transcription factors by binding to the promoter regions of ins1, ins2, and pdx1 genes. In addition, Tub is also bound to the promoter region of the MafA gene. These findings demonstrate that when insulin and leptin resistance develop together in rat PID-MSCs beta-cell differentiation increases markedly via β-catenin and Tub. New therapeutic agents that inhibit AKT/GSK-3β/β-catenin and in particular Tub may help prevent the development or retard the progression of type 2 diabetes.

The members of Acanthamoeba genus are ubiquitous amoeba which could be a pathogenic parasite. The amoeba is resistant to the common chlorine concentration that used for disinfecting the swimming pool water. Therefore, the pools can be suitable environments for the survival and multiplication of the amoeba. In this cross sectional study, 10 indoor recreational water centers from different regions of Tabriz city were selected and sampling was done from fixed and floating biofilms of the swimming pools and hot tubs. The samples were cultured and monitored for the presence of amoeba cyst or trophozoite. For molecular identification of Acanthamoeba, PCR (polymerase chain reaction) and sequencing were conducted based on genus specific fragment of 18S ribosomal DNA (Rns). Acanthamoeba contamination was observed in 6 centers of 10 recreational centers. Based on the amoeba isolation from fixed and floating biofilms, 2 (20%) swimming pools, and 5 (50%) hot tubs were contaminated. Based on the type of the sample, the highest contamination was found in the hot tub water (40%) and the least was found in the swimming pools water (10%) and fixed biofilms of the swimming pools (10%). Out of 8 isolates, 5 (62.5%) were shown expected product in PCR amplification. Sequence analysis showed that Acanthamoeba isolates belonged to the T3 and T4 genotypes. The study revealed a high degree of contamination in the indoor recreational water centers in Tabriz city. So, it is essential to pay closer attention to the hygiene of swimming pools and hot tubs.

Based on type studies and freshly collected material we here re-instate the genus Thyronectria (Nectriaceae, Hypocreales). Species of this genus were recently for the most part classified in the genera Pleonectria (Nectriaceae) or Mattirolia (Thyridiaceae), because Thyronectria and other genera had been identified as members of the Thyridiaceae due to the presence of paraphyses. Molecular phylogenies based on several markers (act, ITS, LSU rDNA, rpb1, rpb2, tef1, tub) revealed that the Nectriaceae contain members whose ascomata are characterised by long, more or less persistent, apical paraphyses. All of these belong to a single genus, Thyronectria, which thus has representatives with hyaline, rosy, green or even dark brown and sometimes distoseptate ascospores. The type species of Thyronectria, T. rhodochlora, syn. T. patavina, syn. T. pyrrhochlora is re-described and illustrated. Within the Nectriaceae persistent, apical paraphyses are common in Thyronectria and rarely also occur in Nectria. The genus Mattirolia is revised and merged with Thyronectria and also Thyronectroidea is regarded as a synonym of Thyronectria. The three new species T. asturiensis, T. caudata and T. obscura are added to the genus. Species recently described in Pleonectria as well as some species of Mattirolia are combined in the genus, and a key to Thyronectria is provided. Five species are epitypified. The type species of the genus Thyridium (Thyridiaceae), T. vestitum, is included in phylogenetic analyses to illustrate the phylogenetic distance of Thyronectria from the Thyridiaceae.

Inherited retinal dystrophies are a major cause of childhood blindness. Here, we describe the identification of a homozygous frameshift mutation (c.1194_1195delAG, p.Arg398Serfs*9) in TUB in a child from a consanguineous UK Caucasian family investigated using autozygosity mapping and whole-exome sequencing. The proband presented with obesity, night blindness, decreased visual acuity, and electrophysiological features of a rod cone dystrophy. The mutation was also found in two of the proband\'s siblings with retinal dystrophy and resulted in mislocalization of the truncated protein. In contrast to known forms of retinal dystrophy, including those caused by mutations in the tubby-like protein TULP-1, loss of function of TUB in the proband and two affected family members was associated with early-onset obesity, consistent with an additional role for TUB in energy homeostasis.

The brown planthopper, Nilaparvata lugens (Stål), is a globally devastating insect pest of rice, particularly in eastern Asia. Distal-less or Dll is a highly conserved and well studied transcription factor required for limb formation in invertebrates and vertebrates. We have identified a homologue of this gene, NlDll, and demonstrated that it is expressed in all life stages of N. lugens, particularly in adult brachypterous females. When we compared between specific adult tissues it was expressed most strongly in wings. Using RNAi techniques we demonstrated that downregulation of NlDll in the 3rd instar larvae led to the disrupted development of the leg, while downregulation of NlDll in the 5th instar larvae led to abnormal wing formation. Ectopic over-expression of NlDll in Drosophila melanogaster using the GAL4-UAS system led to fatal or visible phenotypic changes such as the loss of normal wing structure and disrupted haltere structure. Our work suggests that NlDll is a conserved homologue of Distal-less and is required for both leg development and wing structure. Since researches have shown that Dll is required for wing morphogenesis, understanding the role of NlDll during the wing development will further provide a basis for revealing the molecular mechanism of the wing dimorphism in brown planthopper. In the future, NlDll could be used as a target gene for brown planthopper pest management in the field.