Self-Assembled Oligopeptoplex-Loaded Dissolving Microneedles for Adipocyte-Targeted Anti-Obesity Gene Therapy.

Advancements in gene delivery systems are pivotal for gene-based therapeutics in oncological, inflammatory, and infectious diseases. This study delineates the design of a self-assembled oligopeptoplex (SA-OP) optimized for shRNA delivery to adipocytes, targeting obesity and associated metabolic syndromes. Conventional systems face challenges, including instability due to electrostatic interactions between genetic materials and cationic oligopeptides. Additionally, repeated injections induce discomfort and compromise patient well-being. To circumvent these issues, a dissolvable hyaluronic acid-based, self-locking microneedle (LMN) patch is developed, with improved micro-dose efficiency, for precise SA-OP delivery. This platform offers pain-free administration and improved SA-OP storage stability. In vitro studies in 3T3-L1 cells demonstrated improvements in SA-OP preservation and gene silencing efficacy. In vivo evaluation in a mice model of diet-induced type 2 diabetes yielded significant gene silencing in adipose tissue and a 21.92 ± 2.51% reduction in body weight with minimum relapse risk at 6-weeks post-treatment, representing a superior therapeutic efficacy in a truncated timeframe relative to the GLP-1 analogues currently available on the market. Additionally, SA-OP (LMN) mitigated insulin resistance, inflammation, and hepatic steatosis. These findings establish SA-OP (LMN) as a robust, minimally invasive transdermal gene delivery platform with prolonged storage stability for treating obesity and its metabolic comorbidities.

Developmental Study on "Smart Silver Care": A Mobile Application to Alleviate Loneliness in Older Adults within the Community.

BACKGROUND: Loneliness poses a significant threat to the quality of life of older adults. Therefore, it is essential to implement non-face-to-face services to solve the loneliness of older adults in the community. OBJECTIVES: This study used the ADDIE (Analysis, Design, Development, Implementation, and Evaluation) model to develop mobile applications as a loneliness intervention for older adults living in the community. METHODS: A mobile application was developed using the ADDIE model to alleviate loneliness in older adults living in the community. The development process included a systematic review, a face-to-face preference survey, and an experts' evaluation. From 11 to 15 June 2021, the following six databases were used to search for related articles: Ovid-Medline, Ovid-EMBASE, Cochrane Library, KISS, Korea Med, RISS. A preference analysis was conducted on 100 adults aged 65 or older living in the community from 15 July to 31 August 2021. RESULTS: A mobile application for community-dwelling older adults was developed. Its contents included emotional support, cognition, physical activity, health data, nutrition, and motivation. They were organized through a systematic review and preference survey in the analysis stage. They were also designed as main menus and sub-content at the design stage. They also designed the structure, functionality, and interface layout. The application was developed by integrating the designed content and determining the operating system, language, access method, privacy, and server history. Then, experts evaluated the validity of the application. CONCLUSIONS: The prototype mobile application incorporates emotional support, cognition, physical activity, health data, nutrition, and motivation. It is expected to help older adults achieve their goals by promoting participation. By incorporating expert validity into the content development process of early prototypes, we have improved the usability and acceptability of our products. Future pilot trials are needed to evaluate the effectiveness of this mobile application among older adults.

Incidence and risk factors of infection in a single cohort of 110 adults with systemic lupus erythematosus.

Infection is a major cause of mortality in patients with systemic lupus erythematosus (SLE). This study describes infectious complications in SLE patients and analyzes the risk factors for infection at the time of SLE diagnosis and during the course of SLE in a case-control study. Of 110 patients enrolled, 42 (38%) had at least 1 episode of infectious disease. The incidence of infectious disease was 4.4/100 patient-years (py) with a total follow-up duration of 954 y. In multivariate analysis, independent predictors of infection at the time of SLE diagnosis were an SLE disease activity index (SLEDAI) > 12 (p = 0.01), C3 levels < 90 mg/dl (p = 0.01) and positive anti-ds DNA antibodies (p < 0.01). Frequent flare-ups (p = 0.04) and follow-up duration > or =8 y (p = 0.023) were also significant risk factors for infectious diseases. It is mandatory to closely observe SLE patients with risk factors for developing infectious diseases.

DUB-1A, a novel deubiquitinating enzyme subfamily member, is polyubiquitinated and cytokine-inducible in B-lymphocytes.

Recently, we isolated the Dub-2A gene, which encodes a novel murine deubiquitinating enzyme subfamily member, from a bacterial artificial chromosome library clone by PCR amplification with degenerate PCR primers for the Dub-2 cDNA (Baek, K.-H., Mondoux, M. A., Jaster, R., Fire-Levin E., and D'Andrea, A. D. (2001) Blood 98, 636-642). In this study, we analyzed two more clones from the library to isolate genes encoding other deubiquitinating enzymes. Dub-1A, which encodes the shortest member of the DUB subfamily of deubiquitinating enzymes so far, has been identified in both clones and characterized. Sequence analysis showed that Dub-1A encodes a 468-amino acid protein that has a molecular mass of approximately 51 kDa and that contains a putative catalytic domain (Cys, His, and Asp) conserved among DUB proteins. The amino acid sequence of DUB-1A is 84.5, 84.7, and 85.3% identical to those of DUB-1, DUB-2, and DUB-2A, respectively. Reverse transcription-PCR revealed that Dub-1A is expressed not only in B-lymphocytes in response to interleukin-3 stimulation, but also in T-lymphocytes, brain, heart, liver, lung, kidney, ovary, and spleen. This suggests that Dub-1A may play essential roles in each of these organs. In vivo and in vitro deubiquitinating enzyme assays showed that DUB-1A has functional deubiquitinating activity and that the 5'-flanking sequence of Dub-1A has a functional enhancer domain as shown in Dub-1 and Dub-2A. Interestingly, immunoblot analysis revealed that DUB-1A is polyubiquitinated, indicating that it is degraded through proteasome-mediated degradation. In the absence of JAK2, Dub-1A was expressed at a lower level. This suggests that DUB-1A functions downstream of JAK2 kinase in the interleukin-3 signaling pathway.

Synthesis and evaluation of derivatives of leucine enkephalin as potential affinity labels for delta opioid receptors.

As part of an effort to develop peptide-based affinity labels for opioid receptors, [Leu(5)]enkephalin (LeuEnk) and DTLET (Tyr-D-Thr-Gly-Phe-Leu-Thr), potent agonists for delta receptors, were selected as the parent peptides for further modification. The affinity label derivatives were prepared using standard Fmoc solid-phase peptide synthesis in conjunction with Fmoc-Phe(p-NHAlloc) (Fmoc: 9-flourenylmethoxycarbonyl;) and selective modification of the p-amino group on this residue. The electrophilic isothiocyanate and bromoacetamide groups were introduced into the para position of Phe(4); the corresponding free amine-containing peptides were also prepared for comparison. The pure peptides were evaluated in radioligand binding assays using Chinese hamster ovary (CHO) cells expressing delta and micro opioid receptors. Modification of Phe(4) in LeuEnk and DTLET significantly decreased delta-receptor binding affinity (40 to >2,000-fold). Among the synthesized analogues, [Phe(p-NH(2))(4)]DTLET showed the highest delta-receptor binding affinity (IC(50) = 39 nM) and enhanced selectivity for delta receptors compared to DTLET while other derivatives exhibited much lower delta receptor affinity. The differences in affinities between the two series of analogues and between the derivatives of LeuEnk and N,N-dibenzyl[Leu(5)]Enk reported previously suggest subtle differences in interactions of Phe(4) with delta receptors depending on other modifications in the sequences.

Expression of angiogenesis- and apoptosis-related genes in chorionic villi derived from recurrent pregnancy loss patients.

Angiogenesis takes place during embryogenesis, characterized by the formation of new blood vessels from pre-existing ones. This biological process is also found in the female reproductive system, wound healing, and cancer development. Apoptosis, programmed cell death, is a physiological process in development, tissue homeostasis, and disease. Apoptosis is a normal event in several reproductive tissues including human placenta. In these studies, we investigated whether aberrant angiogenesis and apoptosis are associated with recurrent pregnancy loss (RPL). We compared the gene expression level for angiogenesis- and apoptosis-related genes in chorionic villi from RPL patients and those from normal controls. Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed that 7 angiogenesis- and 12 apoptosis-related genes were abnormally expressed in chorionic villi from RPL patients. Angiogenesis-related genes that showed aberrant expression level are matrix metalloproteinase-2 (MMP-2), plasminogen activator inhibitor (PAI), integrin, transforming growth factor-beta (TGF-beta), vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), and leptin receptor. Expression levels for these genes, except for leptin receptor, showed less in chorionic villi from RPL patients than those from normal controls. In contrast, higher expression levels of 12 apoptosis-related genes (caspase 3, 6, 7, 8, 9, 10, 12, BAD, BAX, BID, Fas, and FasL) were shown in chorionic villi from RPL patients than those from normal controls. Taken all together, it is likely that the lower expression of angiogenesis-related genes and the excessive expression of apoptosis-related genes are associated with RPL.

Molecular cloning and complete cDNA sequence of UBH1 in mouse testis.

We have identified a full-length mouse UBH1 cDNA, encoding a putative deubiquitinating enzyme, from the testis by RT-PCR using primers prepared from sequences conserved amongst deubiquitinating enzymes. Sequence analysis predicts that the UBH1 cDNA encodes a 355 amino acid polypeptide with the molecular weight of approximately 39 kDa containing the highly conserved Cys, Asp, and His domains characteristic of the ubiquitin-specific processing proteases. Biochemical assay revealed that the mouse UBH1 has deubiquitinating enzyme activity and sequence analysis showed 98.3% amino acid identity with human UBH1.

Comparison of gene expression at the feto-maternal interface between normal and recurrent pregnancy loss patients.

Normal pregnancy requires a series of immunological, metabolic, vascular and endocrine regulating processes. However, the specific genes and proteins involved in these processes are not well defined. Aberration of these processes may lead to problems in pregnancy. One of these problems may be recurrent pregnancy loss (RPL). Little information is available on the level of expression of genes that may play a role in normal pregnancy. Therefore, this study determined whether different levels of gene expression at the feto-maternal interface could be associated with factors for RPL. The expression patterns of genes isolated from subtractive hybridization analysis performed with chorionic villi from normal and abnormal pregnancies were investigated. Eight genes classified into groups, including immunosuppression-related, embryo attachment-related and angiogenesis-related, were isolated.

Molecular cloning and characterization of bovine bgl-1, a novel family member of WD-40 repeat-containing lethal giant larvae tumor suppressor genes.

In this study, we have isolated a bovine homologue bgl-1 of lethal giant larvae (lgl) tumor suppressor oncogene from bovine brain by RT-PCR using primers designed based on the conserved sequences for lgl family members. The sequence analysis showed that the bgl-1 encodes a 1,036 amino acid polypeptide with the molecular weight of approximately 112 kDa containing a domain characteristic of WD-40 proteins. The amino acid sequence of bgl-1 showed a homology of 98.3 and 87.3% identity to that of mouse and human, respectively. Northern blot analysis showed that bgl-1 was highly expressed in brain, ovary and testis, with moderate expression in liver, uterus, lung and kidney. This suggests that the bgl-1 may play essential roles in each of these organs. The complementation analysis revealed that the bovine bgl-1 partially restored the Na+ tolerance in the absence of yeast lgl homologue, suggesting that bgl-1 is a bovine homologue of the lgl family.