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1.
Vet Res Forum ; 15(5): 219-222, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39022581

RESUMEN

Foot-and-mouth disease (FMD) is considered as one of the most important contagious viral diseases affecting cloven-footed animals. For effective control of FMD, immunization along with herd immunity is essential in the field conditions. To assure and track the coverage and effectiveness of the vaccination program, the serological studies are very much required after the vaccination program. The present study was aimed to investigate the prevalence of antibodies against structural proteins of FMD virus (FMDV) serotypes of O, A and Asia-1 in seven districts of western Uttar Pradesh, India, and assure the efficacy of vaccination under National Animal Disease Control Program. A total of 308 sera samples were collected from apparent healthy vaccinated cattle and buffaloes from seven districts including Amroha, Baghpat, Bareilly, Bulandsahar, Gautam Budh Nagar, Meerut and Muzaffarnagar of western Uttar Pradesh, India. Determination of antibodies against structural proteins of FMDV was carried out using solid-phase blocking enzyme-linked immunosorbent assay. The protective level of the FMDV serotypes O, A and Asia-1 included in the inactivated trivalent vaccine was 66.55, 48.05 and 47.08% in bovines, respectively. To provide the higher level of protection against the circulating FMDV, the present study recommended the thorough investigation of the immunogenic interaction between the vaccine strains and the field strains. Further investigations should also be conducted with larger sample size and across diverse geographical regions to gain a more comprehensive understanding of herd immunity.

2.
Fitoterapia ; 177: 106123, 2024 Jul 14.
Artículo en Inglés | MEDLINE | ID: mdl-39004288

RESUMEN

The Potentilla genus has long been used traditionally as food and a folklore medicine. In the present study, aerial parts of two Potentilla species, Potentilla fulgens and Potentilla atrosanguinea, of western Himalayan origin, were studied for their anti-breast cancer activity. Ethyl acetate (PAA-EA, PFA-EA), methanolic (PAA-ME, PFA-ME) and hydro-methanolic extract (PAA-HM, PFA-HM) of the plants were tested for their antiproliferative activities against MCF-7 and T-47D breast cancer cell lines. The extracts showed good antiproliferative activity against ER-α dominant breast cancer cell line T-47D, having IC50 values 6.19 ± 0.01 to 33.23 ± 0.04 µg/ml. Eight compounds were isolated, characterized, and quantified from ethyl acetate and methanolic extracts by column chromatography, 1D, 2D-NMR, HRMS and TLC densitometric analysis. Two compounds (4 and 6) have shown better antiproliferative activity than standard bazedoxifene and were further evaluated for their ER-α binding affinity via-fluorescence polarization-based competitive binding assay. The antiestrogenic properties of both compounds were assessed using western blotting. Compounds 4 and 6 were found to have significant affinity for the ER-α and managed to decrease its expression by 38 and 54% respectively. Compounds 4 and 6 also had good stability and reactivity as measured by minimal fluctuations in molecular dynamic simulation analysis, a good dock score in molecular docking, and a respectable HOMO-LUMO energy gap in DFT calculations. Compounds 4 and 6 have shown reliable results and can be used in the development of natural product-based anti-breast cancer agents.

3.
Nat Commun ; 15(1): 5773, 2024 Jul 10.
Artículo en Inglés | MEDLINE | ID: mdl-38982057

RESUMEN

Harnessing the power of cell biocatalysis for sustainable chemical synthesis requires rational integration of living cells with the modern synthetic catalysts. Here, we develop silica-tiling strategy that constructs a hierarchical, inorganic, protocellular confined nanospace around the individual living cell to accommodate molecularly accessible abiotic catalytic sites. This empowers the living microorganisms for new-to-nature chemical synthesis without compromising the cellular regenerative process. Yeast cell, a widely used biocatalyst, is upgraded via highly controlled self-assembly of 2D-bilayer silica-based catalytic modules on cell surfaces, opening the avenues for diverse chemobiotic reactions. For example, combining [AuPt]-catalyzed NADH regeneration, light-induced [Pd]-catalyzed C-C cross-coupling or lipase-catalyzed esterification reactions-with the natural ketoreductase activity inside yeast cell. The conformal silica bilayer provides protection while allowing proximity to catalytic sites and preserving natural cell viability and proliferation. These living nanobiohybrids offer to bridge cell's natural biocatalytic capabilities with customizable heterogeneous metal catalysis, enabling programmable reaction sequences for sustainable chemical synthesis.


Asunto(s)
Biocatálisis , Saccharomyces cerevisiae , Dióxido de Silicio , Dióxido de Silicio/química , Saccharomyces cerevisiae/metabolismo , Catálisis , NAD/metabolismo , Lipasa/metabolismo , Paladio/química , Esterificación
4.
Artículo en Inglés | MEDLINE | ID: mdl-38985420

RESUMEN

Skin homeostasis is predominantly compromised by exposure to UV-B irradiation, leading to several physiopathological processes at cellular and tissue levels that deteriorate skin function and integrity. The current study investigated the photo-protective role of seabuckthorn fruit pulp (SBT) extract against UV-B-induced damage in primary human skin fibroblasts (HDFs) and Balb/C mice skin. We subjected HDFs and Balb/C mice to UV-B irradiation and measured multiple cellular damage indicators. We found that UV-B-irradiated HDFs treated with SBT had a considerably greater survival rate than cells exposed to UV-B radiation alone. The UV-B irradiation-induced ROS generation led to the degradation of the extracellular matrix, inflammation, DNA damage, endoplasmic reticulum (ER) stress, and apoptosis. SBT treatment significantly reduced these manifestations. Topical application of SBT alleviated UV-B-induced epidermal thickening, leukocyte infiltration, and degradation of extracellular matrix in Balb/c mice skin. Based on our results, we conclude that SBT has the potential to be developed as a therapeutic/cosmetic remedy for the prevention of skin photo-damage.

5.
Artículo en Inglés | MEDLINE | ID: mdl-39041320

RESUMEN

Helicobacter pylori, a leading human pathogen associated with duodenal ulcer and gastric cancer, presents a significant threat to human health due to increasing antibiotic resistance rates. This study investigates G-quadruplexes (G4s), which are non-canonical secondary structures form in G-rich regions within the H. pylori genome. Extensive research on G4s in eukaryotes has revealed their role in epigenetically regulating cellular processes like gene transcription, DNA replication, and oncogene expression. However, understanding of G4-mediated gene regulation in other organisms, especially bacterial pathogens, remains limited. Although G4 motifs have been extensively studied in a few bacterial species such as Mycobacterium, Streptococci, and Helicobacter, research on G4 motifs in other bacterial species is still sparse. Like in other organisms such as archaea, mammals, and viruses, G4s in H. pylori display a non-random distribution primarily situated within open reading frames of various protein-coding genes. The occurrence of G4s in functional regions of the genome and their conservation across different species indicates that their placement is not random, suggesting an evolutionary pressure to maintain these sequences at specific genomic sites. Moreover, G-quadruplexes show enrichment in specific gene classes, suggesting their potential involvement in regulating the expression of genes related to cell wall/membrane/envelope biogenesis, amino acid transport, and metabolism. This indicates a probable regulatory role for G4s in controlling the expression of genes essential for H. pylori survival and virulence. Biophysical techniques such as Circular Dichroism spectroscopy and Nuclear Magnetic Resonance were used to characterize G4 motifs within selected H. pylori genes. The study revealed that G-quadruplex ligand inhibited the growth of H. pylori, with minimal inhibitory concentrations in the low micromolar range. This suggests that targeting G4 structures could offer a promising approach for developing novel anti-H. pylori drugs.

6.
J Hazard Mater ; 477: 135234, 2024 Jul 16.
Artículo en Inglés | MEDLINE | ID: mdl-39042990

RESUMEN

Thorium-232 (Th-232) is a promising fuel for advanced nuclear reactors. However, in case of internal human exposure to Th, there is currently no effective modality for its removal from liver and skeleton or for mitigating its effect. The FDA-approved agent, diethylenetriaminepentaacetate (DTPA), can remove Th and other actinides from blood circulation only. For the first time, a rationally-selected polyherbal hepatoprotective i.e. Liv52® (L52S), was evaluated in-combination with DTPA for its Th decorporation ability in Swiss mice. Inductively-coupled plasma mass spectroscopic analysis showed that oral administration of L52S in conjunction with DTPA significantly decreased Th burden from liver (20 %) and skeleton (33 %) as well as enhanced Th excretion (∼2.5 folds) through urine in comparison to DTPA or L52S alone. The combinatorial therapy was found to be complementary in-action, ameliorating Th-induced tissue damage in liver, spleen, and bone more effectively than monotherapy. Furthermore, markers of liver function (alanine transaminase) and liver inflammation and fibrosis (NF-κB & keratin) further validated the beneficial effect of L52S. The human consumption of L52S for various liver disorders further supports its clinical application for Th decorporation and mitigation of its health effects.

8.
Arch Pharm (Weinheim) ; : e2400366, 2024 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-38991221

RESUMEN

The present research focused on the tail-approach synthesis of novel extended thiazolotriazoles (8a-8j) and triazolothiadiazines (11a-11j) including aminotriazole intermediate 10. After successful synthesis, all the compounds were evaluated for their inhibition potential against cytosolic isoforms of human carbonic anhydrase (hCA I, II), tumor-linked transmembrane isoforms (hCA IX, XII), and cathepsin B. As per the inhibition data, the newly synthesized compounds showed poor inhibition against hCA I. Many of the compounds showed effective inhibition toward hCA IX and/or XII in low nanomolar concentration. Despite the strong to moderate inhibition of hCA II by these compounds, more than half of them demonstrated better inhibition against hCA IX and/or XII, comparatively. Further, insights of CA inhibition data of these extended analogs and their comparison with earlier reported thiazolotriazole and triazolothiadiazine derivatives might help in the rational design of novel potent and selective hCA IX and XII inhibitors. The novel compounds were also found to possess anti-cathepsin B potential at a low concentration of 10-7 M. Broadly, compounds of series 11a-11j presented more effective inhibition against cathepsin B than their counterparts in series 8a-8j. Moreover, these in vitro results with respect to cathepsin B inhibition were also supported by the in silico insights obtained via molecular modeling studies.

9.
Polymers (Basel) ; 16(13)2024 Jul 05.
Artículo en Inglés | MEDLINE | ID: mdl-39000773

RESUMEN

This review utilizes an optimized Rouse-Zimm discrete hydrodynamic model and the preaveraged Oseen tensor, which accurately consider hydrodynamic interactions to study model dendrimers. We report the analytical theories that have been previously developed for the creation of generalized analytical models for dendrimers. These generalized theories were used to assess the conformational and dynamical behavior of the dendrimers. By including stiffness in the bonds, the neglect of excluded volume interactions may be somewhat offset. This is true at least in the case of short spacers. While the topological limitations on the directions and orientations of the individual bond vectors in dendrimers implement semiflexibility, the intensity of these contacts was determined by the potential geometric orientations of the bonds, and later on the excluded volume interactions in dendrimers, which were described in terms of the effective co-volume between nearest non-bonded monomers and modeled using the delta function pseudopotential. With the aid of the models developed, the authors condensed various conformational and dynamic properties of dendrimers that depend on their degree of semiflexibility and the strength of the excluded volume. These analyses came to the conclusion that the flexible dendrimer in one limit and the earlier described freely rotating model of dendrimers in the other constitute a highly generalized way of capturing a wide range of conformations in the developed mathematical model in dendrimers.

10.
Cells ; 13(13)2024 Jun 26.
Artículo en Inglés | MEDLINE | ID: mdl-38994961

RESUMEN

Cytokine-induced ß-cell apoptosis is a major pathogenic mechanism in type 1 diabetes (T1D). Despite significant advances in understanding its underlying mechanisms, few drugs have been translated to protect ß-cells in T1D. Epigenetic modulators such as bromodomain-containing BET (bromo- and extra-terminal) proteins are important regulators of immune responses. Pre-clinical studies have demonstrated a protective effect of BET inhibitors in an NOD (non-obese diabetes) mouse model of T1D. However, the effect of BET protein inhibition on ß-cell function in response to cytokines is unknown. Here, we demonstrate that I-BET, a BET protein inhibitor, protected ß-cells from cytokine-induced dysfunction and death. In vivo administration of I-BET to mice exposed to low-dose STZ (streptozotocin), a model of T1D, significantly reduced ß-cell apoptosis, suggesting a cytoprotective function. Mechanistically, I-BET treatment inhibited cytokine-induced NF-kB signaling and enhanced FOXO1-mediated anti-oxidant response in ß-cells. RNA-Seq analysis revealed that I-BET treatment also suppressed pathways involved in apoptosis while maintaining the expression of genes critical for ß-cell function, such as Pdx1 and Ins1. Taken together, this study demonstrates that I-BET is effective in protecting ß-cells from cytokine-induced dysfunction and apoptosis, and targeting BET proteins could have potential therapeutic value in preserving ß-cell functional mass in T1D.


Asunto(s)
Apoptosis , Citocinas , Células Secretoras de Insulina , FN-kappa B , Transducción de Señal , Animales , Células Secretoras de Insulina/efectos de los fármacos , Células Secretoras de Insulina/metabolismo , Células Secretoras de Insulina/patología , FN-kappa B/metabolismo , Ratones , Citocinas/metabolismo , Transducción de Señal/efectos de los fármacos , Apoptosis/efectos de los fármacos , Diabetes Mellitus Experimental/patología , Diabetes Mellitus Experimental/metabolismo , Diabetes Mellitus Tipo 1/metabolismo , Diabetes Mellitus Tipo 1/patología , Proteína Forkhead Box O1/metabolismo , Ratones Endogámicos NOD , Masculino , Ratones Endogámicos C57BL
11.
Chem Commun (Camb) ; 2024 Jul 16.
Artículo en Inglés | MEDLINE | ID: mdl-39011868

RESUMEN

Herein we describe the rational design, synthesis and photophysical study of a novel class of phenanthridine-based, one- and two-photon sensitive, photoremovable protecting groups with absorption wavelengths extending beyond 400 nm. This design facilitated the development of scaffolds with enhanced uncaging quantum yield, paving the way for broader applications in controlled drug delivery and molecular manipulation.

12.
Ann Afr Med ; 23(3): 482-487, 2024 Jul 01.
Artículo en Francés, Inglés | MEDLINE | ID: mdl-39034576

RESUMEN

BACKGROUND: When the range of motion between two finger segments, both active and passive, is restricted, finger contracture occurs. The aim of this study is to investigate the best procedure to eliminate finger contracture and the functional and esthetic results of the different surgical procedures. MATERIALS AND METHODS: A total of 31 patients with soft-tissue abnormalities of the hand were included in this prospective study. They underwent either contracture removal with K-wire and skin grafts or various flap procedures in the department of plastic surgery. Complaints of stiffness and discomfort were classified into five categories: none, mild, moderate, marked, and severe. The difficulty a person had in picking up objects, grasping, writing, etc., was used to determine the degree of disability. Absenteeism from work and surgical site infections were also recorded. RESULTS: The mean age was 20.25 years, with a mean age of 23.05 for men and 15.83 for women. Overall, most cases occurred in the age range of 3-10 years. For K-wire surgery with skin grafting, the typical time off work was 24 days. The average recovery time ranged from 15.2 days for skin grafts to 16.9 days for tenolysis, 28.33 days for groyne flaps, and 41 days for abdominal flaps. Of all cases, 12 (38.00%) had a fair result, 10 (31.04%) had a moderate result, and 9 (30.96%) had an excellent result. CONCLUSION: The most feasible method for treating these situations, which offers the greatest potential for a functional and cosmetic result, is contracture reduction with skin grafting.


Résumé Contexte:Lorsque l'amplitude de mouvement entre deux segments de doigts, actifs et passifs, est restreinte, une contracture des doigts se produit. Le Le but de cette étude est d'étudier la meilleure procédure pour éliminer la contracture des doigts et les résultats fonctionnels et esthétiques des différents interventions chirurgicales.Matériels et méthodes:Au total, 31 patients présentant des anomalies des tissus mous de la main ont été inclus dans cette étude prospective. étude. Ils ont subi soit une ablation des contractures avec du fil K et des greffes de peau, soit diverses procédures de lambeau dans le service de chirurgie plastique. Les plaintes de raideur et d'inconfort ont été classées en cinq catégories: aucune, légère, modérée, marquée et grave. La difficulté d'une personne qu'ils avaient à ramasser des objets, à les saisir, à écrire, etc., a été utilisé pour déterminer le degré d'incapacité. Absentéisme au travail et sur le site chirurgical des infections ont également été enregistrées.Résultats:L'âge moyen était de 20,25 ans, avec un âge moyen de 23,05 ans pour les hommes et de 15,83 ans pour les femmes. Dans l'ensemble, la plupart des cas sont survenus dans la tranche d'âge de 3 à 10 ans. Pour la chirurgie au fil K avec greffe de peau, le temps d'arrêt typique était de 24 jours. La moyenne le temps de récupération variait de 15,2 jours pour les greffes de peau à 16,9 jours pour la ténolyse, 28,33 jours pour les lambeaux d'épi et 41 jours pour les lambeaux abdominaux. Parmi tous les cas, 12 (38,00 %) ont eu un résultat passable, 10 (31,04 %) ont eu un résultat modéré et 9 (30,96 %) ont eu un excellent résultat.Conclusion:le plus La méthode réalisable pour traiter ces situations, qui offre le plus grand potentiel de résultat fonctionnel et esthétique, est la réduction des contractures. avec greffe de peau.


Asunto(s)
Contractura , Trasplante de Piel , Colgajos Quirúrgicos , Centros de Atención Terciaria , Humanos , Femenino , Masculino , Estudios Prospectivos , Contractura/cirugía , Adulto , Niño , Adolescente , Trasplante de Piel/métodos , Preescolar , Adulto Joven , Resultado del Tratamiento , Rango del Movimiento Articular , Dedos/cirugía , Procedimientos de Cirugía Plástica/métodos , Persona de Mediana Edad , Hilos Ortopédicos
13.
Adv Cancer Res ; 161: 321-365, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-39032953

RESUMEN

Prostate cancer (PC) has a high propensity to develop bone metastases, causing severe pain and pathological fractures that profoundly impact a patients' normal functions. Current clinical intervention is mainly palliative focused on pain management, and tumor progression is refractory to standard therapeutic regimens. This limited treatment efficacy is at least partially due to a lack of comprehensive understanding of the molecular landscape of the disease pathology, along with the intensive overlapping of physiological and pathological molecular signaling. The niche is overwhelmed with diverse cell types with inter- and intra-heterogeneity, along with growth factor-enriched cells that are supportive of invading cell proliferation, providing an additional layer of complexity. This review seeks to provide molecular insights into mechanisms underlying PC bone metastasis development and progression.


Asunto(s)
Neoplasias Óseas , Neoplasias de la Próstata , Humanos , Neoplasias Óseas/secundario , Neoplasias Óseas/patología , Neoplasias Óseas/metabolismo , Neoplasias de la Próstata/patología , Neoplasias de la Próstata/genética , Masculino , Animales , Microambiente Tumoral , Transducción de Señal
14.
ACS Catal ; 14(14): 10624-10634, 2024 Jul 19.
Artículo en Inglés | MEDLINE | ID: mdl-39050896

RESUMEN

We report here a method of making polyketones from the coupling of diketones and diols using a manganese pincer complex. The methodology allows us to access various polyketones (polyarylalkylketone) containing aryl, alkyl, and ether functionalities, bridging the gap between the two classes of commercially available polyketones: aliphatic polyketones and polyaryletherketones. Using this methodology, 12 polyketones have been synthesized and characterized using various analytical techniques to understand their chemical, physical, morphological, and mechanical properties. Based on previous reports and our studies, we suggest that the polymerization occurs via a hydrogen-borrowing mechanism that involves the dehydrogenation of diols to dialdehyde followed by aldol condensation of dialdehyde with diketones to form chalcone derivatives and their subsequent hydrogenation to form polyarylalkylketones.

15.
Nat Commun ; 15(1): 6253, 2024 Jul 24.
Artículo en Inglés | MEDLINE | ID: mdl-39048574

RESUMEN

We report here a method to make a branched and partially ethoxylated polyethyleneimine derivative directly from ethanolamine. The polymerization reaction is catalysed by a pincer complex of Earth-abundant metal, manganese, and produces water as the only byproduct. Industrial processes to produce polyethyleneimines involve the transformation of ethanolamine to a highly toxic chemical, aziridine, by an energy-intensive/waste-generating process followed by the ring-opening polymerization of aziridine. The reported method bypasses the need to produce a highly toxic intermediate and presents advantages over the current state-of-the-art. We propose that the polymerization process follows a hydrogen borrowing pathway that involves (a) dehydrogenation of ethanolamine to form 2-aminoacetaldehyde, (b) dehydrative coupling of 2-aminoacetaldehyde with ethanolamine to form an imine derivative, and (c) subsequent hydrogenation of imine derivative to form alkylated amines.

16.
J Hazard Mater ; 476: 135073, 2024 Jul 03.
Artículo en Inglés | MEDLINE | ID: mdl-38968826

RESUMEN

This study conducted a comprehensive analysis of trace element concentrations in the Upper Indus River Basin (UIRB), a glacier-fed region in the Western Himalayas (WH), aiming to discern their environmental and anthropogenic sources and implications. Despite limited prior data, 69 samples were collected in 2019 from diverse sources within the UIRB, including mainstream, tributaries, and groundwater, to assess trace element concentrations. Enrichment factor (EF) results and comparisons with regional and global averages suggest that rising levels of Zn, Cd, and As may pose safety concerns for drinking water quality. Advanced multivariate statistical techniques such as principal component analysis (PCA), absolute principal component scores (APCS-MLR), Monte Carlo simulation (MCS), etc were applied to estimate the associated human health hazards and also identified key sources of trace elements. The 95th percentile of the MCS results indicates that the estimated total cancer risk for children is significantly greater than (>1000 times) the USEPA's acceptable risk threshold of 1.0 × 10-6. The results classified most of the trace elements into two distinct groups: Group A (Li, Rb, Sr, U, Cs, V, Ni, TI, Sb, Mo, Ge), linked to geogenic sources, showed lower concentrations in the lower-middle river reaches, including tributaries and downstream regions. Group B (Pb, Nb, Cr, Zn, Be, Al, Th, Ga, Cu, Co), influenced by both geogenic and anthropogenic activities, exhibited higher concentrations near urban centers and midstream areas, aligning with increased municipal waste and agricultural activities. Furthermore, APCS-MLR source apportionment indicated that trace elements originated from natural geogenic processes, including rock-water interactions and mineral dissolution, as well as anthropogenic activities. These findings underscore the need for targeted measures to mitigate anthropogenic impacts and safeguard water resources for communities along the IRB and WH.

17.
World J Orthop ; 15(6): 570-577, 2024 Jun 18.
Artículo en Inglés | MEDLINE | ID: mdl-38947270

RESUMEN

BACKGROUND: The preferred treatment for distal humeral intercondylar fractures is open reduction and internal fixation. While there is consensus about the posterior approach, several posterior approaches have been developed. It is debatable as to which approach is best. AIM: To compare triceps reflecting anconeus pedicle (TRAP) and olecranon osteotomy approaches for internal fixation of distal humeral intercondylar fracture. METHODS: In total, 40 cases of Arbeitsgemeinschaft für Osteosynthesefragen/Association of the Study of Internal Fixation type C, closed, and Gustilo type I intercondylar humeral fractures were included. Patients ranged in age from 18 years to 70 years. The patients were randomized into two groups: TRAP group and olecranon osteotomy group, with 20 cases in each. All were followed up at 6 wk, 3 months, 6 months, and 12 months. Functional outcomes were measured in terms of flexion-extension arc, Disabilities of Arm, Shoulder and Hand score, and Mayo Elbow Performance Score. RESULTS: The mean age was 43.2 years in the TRAP group and 37.5 years in the olecranon osteotomy group. The mean operative time and mean duration of hospital stay in the TRAP group were significantly higher than in the olecranon osteotomy group (119.5 vs 111.5 min and 9.85 vs 5.45 d, respectively). The mean arc of flexion-extension, Disabilities of Arm, Shoulder and Hand score, and Mayo Elbow Performance Score were comparable without any significant difference in the groups at the 12-month follow-up (107.0 vs 106.2, 18.3 vs 15.7, and 84.2 vs 86.2, respectively). Ulnar paresthesia and superficial infections were comparable in both groups (2 cases vs 3 cases and 3 cases vs 2 cases, respectively). Hardware prominence was significantly higher in the olecranon osteotomy group, mostly due to tension band wiring. CONCLUSION: Both approaches were equivalent, but there is a need for further study including higher numbers of subjects and longer study duration to prove the benefits of one approach over the other.

18.
Nat Struct Mol Biol ; 2024 Jul 01.
Artículo en Inglés | MEDLINE | ID: mdl-38951623

RESUMEN

The development of precise RNA-editing tools is essential for the advancement of RNA therapeutics. CRISPR (clustered regularly interspaced short palindromic repeats) PspCas13b is a programmable RNA nuclease predicted to offer superior specificity because of its 30-nucleotide spacer sequence. However, its design principles and its on-target, off-target and collateral activities remain poorly characterized. Here, we present single-base tiled screening and computational analyses that identify key design principles for potent and highly selective RNA recognition and cleavage in human cells. We show that the de novo design of spacers containing guanosine bases at precise positions can greatly enhance the catalytic activity of inefficient CRISPR RNAs (crRNAs). These validated design principles (integrated into an online tool, https://cas13target.azurewebsites.net/ ) can predict highly effective crRNAs with ~90% accuracy. Furthermore, the comprehensive spacer-target mutagenesis revealed that PspCas13b can tolerate only up to four mismatches and requires ~26-nucleotide base pairing with the target to activate its nuclease domains, highlighting its superior specificity compared to other RNA or DNA interference tools. On the basis of this targeting resolution, we predict an extremely low probability of PspCas13b having off-target effects on other cellular transcripts. Proteomic analysis validated this prediction and showed that, unlike other Cas13 orthologs, PspCas13b exhibits potent on-target activity and lacks collateral effects.

19.
J Toxicol Pathol ; 37(3): 109-126, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38962260

RESUMEN

The historical control database of a multinational laboratory services provider was queried for all histopathologic findings in New Zealand White rabbits which were used as control animals during a ten-year period (2011-2020). The query included all evaluated tissues, with or without microscopic findings, in studies conducted for safety testing for regulatory approval by the U.S. Food and Drug Agency (FDA) or the U.S. Environmental Protection Agency. A second query included studies conducted in the United Kingdom for control rabbits used in studies compliant with the Healthcare Products Regulatory Agency (MHRA) and/or the European Medicines Agency (EMA), which provide regulatory oversight in the United Kingdom and European Union, respectively. Infiltrates of inflammatory (mixed or mononuclear) cells were commonly noted in various organs including heart, digestive tract, muscle, thyroid, kidney, urinary bladder, eyelid, ocular structures, harderian gland, lacrimal gland, and lung. Mineralization was noted in aorta, kidney, urinary bladder, and ovary. Also noted were degeneration/necrosis in the myocardium, and intramuscular injection sites of the skin, degeneration/regeneration of muscle and diaphragm, ectopic tissue in the pancreas and thyroid, basophilic foci in salivary gland, increased/decreased vacuolation in adrenal gland, increased/decreased lymphocytic cellularity of lymph nodes, intrasinusoidal erythrocytes in lymph nodes, thymic atrophy, increased adipocytes in bone marrow, inflammatory cell foci in the liver and gall bladder, lacrimal gland atrophy, renal tubule basophilia, degeneration/regeneration, and dilatation; oviduct cyst; in the testis, degeneration/atrophy, cellular debris, dilatation, decreased sperm and segmental hypoplasia of seminiferous tubules; and squamous metaplasia of the testis and seminal vesicle.

20.
Cureus ; 16(6): e61510, 2024 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-38962593

RESUMEN

Plateletpheresis has become a pivotal part of transfusion medicine. With the increasing demand for plateletpheresis, donor safety is an area of concern because plateletpheresis alters donor hematological parameters. For a better understanding of plateletpheresis, a systemic review is needed to study more evidence-based aspects of plateletpheresis. Electronic databases PubMed, Google Scholar, and Cochrane Library were used to find articles from January 1, 1980, to May 23, 2024. The random effect model was used to meta-analyze the effect of plateletpheresis on hematocrit, hemoglobin, and red blood cell (RBC) count. The Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guideline was followed. A total of 24 studies were found; the effect of plateletpheresis on hemoglobin, hematocrit, and RBC count was studied in the following respective numbers of donors: 3,374, 3,374, and 690. A decrease of hemoglobin, hematocrit, and RBC count was observed after plateletpheresis having a weighted mean difference (WMD) of 0.50 (95%CI = -0.72 to -0.27), WMD of -1.36 (95%CI = -2.05 to -0.66), and WMD of -0.18 (95%CI = -0.23 to -0.12), respectively. Plateletpheresis shows a decrease in the value of hematological parameters such as hemoglobin, hematocrit, and erythrocyte count due to blood loss in the kits employed in the procedure; cell lysis was also seen because of exposure of erythrocytes to stress or change in osmotic pressure. Thus, strict criteria for donation must be developed for better safety of the donors. Improved automated cell separators for plateletpheresis should be made available in blood banks to ensure good quality hematologic products. Our findings suggest that the duration of the procedure should be decreased.

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