KHARON Is an essential cytoskeletal protein involved in the trafficking of flagellar membrane proteins and cell division in African trypanosomes

African trypanosomes and related kinetoplastid parasites selectively traffic specific membrane proteins to the flagellar membrane, but the mechanisms for this trafficking are poorly understood. We show here that KHARON, a protein originally identified in Leishmania parasites, interacts with a putative trypanosome calcium channel and is required for its targeting to the flagellar membrane. KHARON is located at the base of the flagellar axoneme, where it likely mediates targeting of flagellar membrane proteins, but is also on the subpellicular microtubules and the mitotic spindle. Hence, KHARON is probably a multifunctional protein that associates with several components of the trypanosome cytoskeleton. RNA interference-mediated knockdown of KHARON mRNA results in failure of the calcium channel to enter the flagellar membrane, detachment of the flagellum from the cell body, and disruption of mitotic spindles. Furthermore, knockdown of KHARON mRNA induces a lethal failure of cytokinesis in both bloodstream (mammalian host) and procyclic (insect vector) life cycle stages, and KHARON is thus critical for parasite viability

Internally Controlled, Generic Real-Time PCR for Quantification and Multiplex Real-Time PCR with Serotype-Specific Probes for Serotyping of Dengue Virus Infections

Dengue has become a global public health problem and a sensitive diagnostic test for early phase detection can be life saving. An internally controlled, generic real-time PCR was developed and validated by testing serial dilutions of a DENV positive control RNA in the presence of a fixed amount of IC with results showing a good linearity (R2 = 0.9967) and a LOD of at least 1.95 × 104 copies/mL. Application of the generic PCR on 136 patient samples revealed a sensitivity of 95.8% and specificity of 100%. A newly developed multiplex real-time PCR with serotype-specific probes allowed the serotyping of DENV for 80 out of 92 (87%) generic real-time PCR positive patients. Combined these real-time PCRs offer a convenient diagnostic tool for the sensitive and specific quantification of DENV in clinical specimens with the possibility for serotyping

The accuracy of chromosomal microarray testing for identification of embryonic mosaicism in human blastocysts

A dança é uma forma ancestral de magia, invenção dos deuses que a ensinaram aos homens, diz-nos a mitologia hindu. Envolvido no mistério e movimento da dança, o dançarino pode encantar; porém, antes de tudo é preciso que encante a si mesmo. Não seria este também o caminho do professor? Fazer para si para poder fazer ou propor aos educandos, encantar-se para poder encantar; criar para poder seguir com as crianças a aventura da criação; ousar para poder encorajar? Nesta direção, a pergunta que percorre o presente artigo é assim formulada: como contribuir com o processo de encantamento dos professores, como alimentar a sensibilidade, nos percursos da formação universitária? Buscando respostas no processo de pesquisa, identifica-se na experiência com as danças circulares, tradição de diferentes povos, um profícuo caminho pelo qual aquele espaço de encantamento, de inteireza, de educação estética, igualmente, pode ser provocado

Controllable Group Choreography Using Contrastive Diffusion

Music-driven group choreography poses a considerable challenge but holds significant potential for a wide range of industrial applications. The ability to generate synchronized and visually appealing group dance motions that are aligned with music opens up opportunities in many fields such as entertainment, advertising, and virtual performances. However, most of the recent works are not able to generate high-fidelity long-term motions, or fail to enable controllable experience. In this work, we aim to address the demand for high-quality and customizable group dance generation by effectively governing the consistency and diversity of group choreographies. In particular, we utilize a diffusion-based generative approach to enable the synthesis of flexible number of dancers and long-term group dances, while ensuring coherence to the input music. Ultimately, we introduce a Group Contrastive Diffusion (GCD) strategy to enhance the connection between dancers and their group, presenting the ability to control the consistency or diversity level of the synthesized group animation via the classifier-guidance sampling technique. Through intensive experiments and evaluation, we demonstrate the effectiveness of our approach in producing visually captivating and consistent group dance motions. The experimental results show the capability of our method to achieve the desired levels of consistency and diversity, while maintaining the overall quality of the generated group choreography.</jats:p

Memantine, Simvastatin, and Epicatechin Inhibit 7-Ketocholesterol-induced Apoptosis in Retinal Pigment Epithelial Cells But Not Neurosensory Retinal Cells In Vitro

Purpose: 7-ketocholesterol (7kCh), a natural byproduct of oxidation in lipoprotein deposits, is implicated in the pathogenesis of diabetic retinopathy and age-related macular degeneration (AMD). This study was performed to investigate whether several clinical drugs can inhibit 7kCh-induced caspase activation and mitigate its apoptotic effects on retinal cells in vitro. Method: Two populations of retinal cells, human retinal pigment epithelial cells (ARPE-19) and rat neuroretinal cells (R28), were exposed to 7kCh in the presence of the following inhibitors: Z-VAD-FMK (pan-caspase inhibitor), simvastatin, memantine, epicatechin, and Z-IETD-FMK (caspase-8 inhibitor) or Z-ATAD-FMK (caspase-12 inhibitor). Caspase-3/7, -8, and -12 activity levels were measured by fluorochrome caspase assays to quantify cell death. IncuCyte live-cell microscopic images were obtained to quantify cell counts. Results: Exposure to 7kCh for 24 hours significantly increased caspase activities for both ARPE-19 and R28 cells (P &lt; 0.05). In ARPE cells, pretreatment with various drugs had significantly lower caspase-3/7, -8, and -12 activities, reported in % change in mean signal intensity (msi): Z-VAD-FMK (48% decrease, P &lt; 0.01), memantine (decreased 47.8% at 1 μM, P = 0.0039 and 81.9% at 1 mM, P &lt; 0.001), simvastatin (decreased 85.3% at 0.01 μM, P &lt; 0.001 and 84.8% at 0.05 μM , P &lt; 0.001) or epicatechin (83.6% decrease, P &lt; 0.05), Z-IETD-FMK (68.1% decrease, P &lt; 0.01), and Z-ATAD-FMK (47.7% decrease, P = 0.0017). In contrast, R28 cells exposed to 7kCh continued to have elevated caspase- 3/7, -8, and -12 activities (between 25.7% decrease and 17.5% increase in msi, P &gt; 0.05) regardless of the pretreatment. Conclusion: Several current drugs protect ARPE-19 cells but not R28 cells from 7kChinduced apoptosis, suggesting that a multiple-drug approach is needed to protect both cells types in various retinal diseases

Radical-Initiated Brown Carbon Formation in Sunlit Carbonyl–Amine–Ammonium Sulfate Mixtures and Aqueous Aerosol Particles

Brown carbon (BrC) formed from glyoxal+ammonium sulfate (AS) and methylglyoxal+AS reactions photobleaches quickly, leading to the assumption that BrC formed overnight by Maillard reactions will be rapidly destroyed at sunrise. Here, we tested this assumption by reacting glyoxal, methylglyoxal, glycolaldehyde, or hydroxyacetone in aqueous mixtures with reduced nitrogen species at pH 4–5 in the dark and in sunlight (\u3e350 nm) for at least 10 h. The absorption of fresh carbonyl+AS mixtures decreased when exposed to sunlight, and no BrC formed, as expected from previous work. However, the addition of amines (either methylamine or glycine) allowed BrC to form in sunlight at comparable rates as in the dark. Hydroxyacetone+amine+AS aqueous mixtures generally browned faster in sunlight than in the dark, especially in the presence of HOOH, indicating a radical-initiated BrC formation mechanism is involved. In experiments with airborne aqueous aerosol containing AS, methylamine, and glyoxal or methylglyoxal, browning was further enhanced, especially in sunlight (\u3e300 nm), forming aerosol with optical properties similar to “very weak” atmospheric BrC. Liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) analysis of aerosol filter extracts indicates that exposure of methylglyoxal+AS aqueous aerosol to methylamine gas, sunlight, and cloud processing increases incorporation of ammonia, methylamine, and photolytic species (e.g., acetyl radicals) into conjugated oligomer products. These results suggest that when amines are present, photolysis of first-generation, “dark reaction” BrC (imines and imidazoles) initiates faster, radical-initiated browning processes that may successfully compete with photobleaching, are enhanced in aqueous aerosol particles relative to bulk liquid solutions, and can produce BrC consistent with atmospheric observations

Malignant Precursor Cells Pre-Exist in Human Breast DCIS and Require Autophagy for Survival

BACKGROUND: While it is accepted that a majority of invasive breast cancer progresses from a ductal carcinoma in situ (DCIS) precursor stage, very little is known about the factors that promote survival of DCIS neoplastic cells within the hypoxic, nutrient deprived intraductal microenvironment. METHODOLOGY AND PRINCIPAL FINDINGS: We examined the hypothesis that fresh human DCIS lesions contain pre-existing carcinoma precursor cells. We characterized these cells by full genome molecular cytogenetics (Illumina HumanCytoSNP profile), and signal pathway profiling (Reverse Phase Protein Microarray, 59 endpoints), and demonstrated that autophagy is required for survival and anchorage independent growth of the cytogenetically abnormal tumorigenic DCIS cells. Ex vivo organoid culture of fresh human DCIS lesions, without enzymatic treatment or sorting, induced the emergence of neoplastic epithelial cells exhibiting the following characteristics: a) spontaneous generation of hundreds of spheroids and duct-like 3-D structures in culture within 2-4 weeks; b) tumorigenicity in NOD/SCID mice; c) cytogenetically abnormal (copy number loss or gain in chromosomes including 1, 5, 6, 8, 13, 17) compared to the normal karyotype of the non-neoplastic cells in the source patient's breast tissue; d) in vitro migration and invasion of autologous breast stroma; and e) up-regulation of signal pathways linked to, and components of, cellular autophagy. Multiple autophagy markers were present in the patient's original DCIS lesion and the mouse xenograft. We tested whether autophagy was necessary for survival of cytogenetically abnormal DCIS cells. The lysosomotropic inhibitor (chloroquine phosphate) of autophagy completely suppressed the generation of DCIS spheroids/3-D structures, suppressed ex vivo invasion of autologous stroma, induced apoptosis, suppressed autophagy associated proteins including Atg5, AKT/PI3 Kinase and mTOR, eliminated cytogenetically abnormal spheroid forming cells from the organ culture, and abrogated xenograft tumor formation. CONCLUSIONS: Cytogenetically abnormal spheroid forming, tumorigenic, and invasive neoplastic epithelial cells pre-exist in human DCIS and require cellular autophagy for survival

Cross-Sectional Area of the Longus Colli and Cervical Degenerative Spondylolisthesis: A Retrospective Review Evaluating This Anatomic Consideration

Background: Limited literature is available to define the impact of the longus colli muscle, a deep flexor of the spine, on cervical spine stability despite its close proximity to the vertebrae. Aims and Objectives: The purpose of this study was to determine if longus colli cross-sectional area (CSA) is associated with the severity preoperative cervical degenerative spondylolisthesis. Materials and Methods: Patients undergoing elective anterior cervical discectomy and fusion (ACDF) for cervical spondylolisthesis between 2010-2021 were retrospectively identified. Longus colli cross-sectional areas (CSA) were measured from preoperative MRI images at the C5 level. Preoperative spondylolisthesis measurements were recorded with cervical radiographs. Patients were grouped by quartiles respectively according to longus colli CSAs. Statistical tests compared patient demographics, surgical characteristics, and surgical outcomes between groups. Multiple linear regression analysis was utilized to assess if longus colli CSA predicted cervical spondylolisthesis. Results: A total of 157 patients met inclusion criteria. Group 1 (first quartile) was the oldest (60.4 ± 12.0 years, P = 0.024) and was predominantly female (59.0%, P = 0.001). Group 1 also had the highest maximum spondylolisthesis (0.19 mm, P = 0.031) and highest proportion of grade 2 spondylolisthesis (23.1%, P = 0.003). On regression analysis, lowest quartile of longus colli CSA was an independent predictor of larger measured maximum spondylolisthesis (β: 0.04, P = 0.012). Conclusion: Smaller longus colli CSA is independently associated with a higher grade and degree of preoperative cervical spondylolisthesis, but this finding does not result in adverse postsurgical outcomes

Quantifying the Emergence of Dengue in Hanoi, Vietnam: 1998–2009

Dengue is the most common vector-borne viral disease of humans, causing an estimated 50 million cases per year. The number of countries affected by dengue has increased dramatically in the last 50 years and dengue is now a major public health problem in large parts of the tropical and subtropical world. It is of considerable importance to understand the factors that determine how dengue becomes newly established in areas where the risk of dengue was previously small. Hanoi in North Vietnam is a large city where dengue appears to be emerging. We analyzed 12 years of dengue surveillance data in order to characterize the temporal and spatial epidemiology of dengue in Hanoi and to establish if dengue incidence has been increasing. After excluding the two major outbreak years of 1998 and 2009 and correcting for changes in population age structure over time, we found there was a significant annual increase in the incidence of notified dengue cases over the period 1999–2008. Dengue cases were concentrated in young adults in the highly urban central areas of Hanoi. This study indicates that dengue transmission is increasing in Hanoi and provides a platform for further studies of the underlying drivers of this emergence