Cytoplasmic Dynein and Dynactin Coordinate Microtubule Dynamics at the Growing Plus End

Microtubules are cytoskeletal polymers that serve as long-distance tracks for intracellular transport. However, microtubules are not static tracks; they undergo dynamic instability, a non-equilibrium behavior which allows them to continually remodel. Microtubules must remodel to adapt to the changing needs of the cell, yet they must also be stable enough to allow long-distance transport by kinesins and by cytoplasmic dynein and its partner complex dynactin. Microtubule dynamics are strongly influenced by microtubule-associated proteins, which bind to microtubules and bias them toward growth or shrinkage. A pool of dynein and dynactin localizes to the microtubule plus end and to the cortex, where we hypothesized that they might not be solely poised to initiate retrograde runs, but might also act to modify microtubule dynamics. To test this, we reconstituted microtubule dynamic instability in vitro and visualized it using TIRF microscopy. We show that dynein, when immobilized in an orientation recapitulating its membrane recruitment, can tether growing microtubules, reducing their lateral diffusion and delaying catastrophe. This effect does not represent mere microtubule binding; dynein tethers microtubules more effectively than kinesin or the plus end-tracking protein EB1, and we show that this ability of dynein is dependent on its ATPase activity. Modeling suggests that dynein may delay catastrophe by actively applying tension to straighten, and thus stabilize the microtubule. We also have examined the effects of the p150Glued subunit of dynactin on microtubule dynamics. We find that to modify dynamics, p150Glued must be dimerized, and it must bind tubulin, an interaction that requires its CAP-Gly and basic domains in tandem. p150Glued is alternatively spliced in vivo, with the isoform including both these domains expressed primarily in neurons. Accordingly, depletion of p150Glued in a non-polarized cell line does not alter microtubule dynamics, while p150Glued RNAi in neurons leads to a dramatic increase in microtubule catastrophe. Strikingly, a Parkinson syndrome-associated mutation blocks this microtubule-stabilizing activity both in vitro and in neurons. Together, these data provide novel mechanistic insight into how cytoplasmic dynein and dynactin, the principle minus end-directed motor complex in the cell, also act at the plus end to coordinate microtubule dynamics

A natural hybrid of a Bacillus thuringiensis Cry2A toxin implicates Domain I in specificity determination

A PCR-RFLP method was used to identify cry2A toxin genes in a collection of 300 strains of Bacillus thuringiensis. From 81 genes identified, the vast majority appeared to be cry2Aa or cry2Ab, however three showed a different pattern and were subsequently cloned and sequenced. The gene cloned from strain HD395 was named cry2Ba2. Since the proteins encoded by the genes cloned from LS5115-3 and DS415 shared >95% sequence identity with existing toxins their genes were named cry2Aa17 and cry2Ab29 respectively by the toxin nomenclature committee. Despite this overall similarity these two toxins resembled natural hybrids, with Cry2Ab29 resembling Cry2Ab for the majority of the protein but then showing identity to Cry2Aa for the last 66 amino acids. For Cry2Aa17, Domains II and III most closely resembled Cry2Aa (99% identity) whilst Domain I was identical to that of Cry2Ab. The toxicity of the recombinant toxins was tested against Aedes aegypti and Spodoptera exigua, and it was found that the toxicity profile of Cry2Aa17 more closely matched the profile of Cry2Ab than that of Cry2Aa, thus implicating Domain I in specificity determination. This association of Domain I with toxicity was confirmed when hybrids were made between Cry2Aa and Cry2Ab

Efficacy of cling film dressing on palmoplantar psoriatic lesions among patients with psoriasis: A randomized controlled trial

Introduction: Caring for patients with skin disorders is “more than skin deep”. Psoriasis is a systematic immune-mediated disease that generally does not affect survival, but certainly has major negative effects on patients. The previous research has reported that artificial restoration of a permeable barrier by occlusion results in regression of lesions in psoriasis.The aim of this research is to assess the efficacy of cling film dressing versus conventional treatment on palmoplantar psoriatic lesions among patients with psoriasis. Methods: The research design used in the study was a true experimental time series design. Block randomization was used to assign the study participants into either study or control group in a Psoriatic Out-Patient Department of tertiary care center at Southern part of India. As usual, the intervention group participants received the application of topical corticosteroid Eczivate MF with cling film wrap while the control group participants followed the conventional treatment (topical corticosteroid Eczivate MF without occlusive dressing). The modified psoriasis severity index (MPSI) score was used to measure the outcomes. Results: There was a high statistical significance difference on the severity of psoriatic disease level between the control and study groups (p ˂ 0.05). Conclusion: Steroidal cream application with occlusive dressing yielded better results than non-occlusive open treatment

Predictors of Quality of Life among Omani Family Caregivers for Traumatic Brain Injury Patients

Objective: After acute care, the burden of caring for patients with traumatic brain injury (TBI) is mainly shouldered by the family caregivers (FCs). We aimed to explore the quality of life (QoL) of Omani FCs for patients with TBI. Methods: A total of 36 FCs and patients with TBI were recruited from one hospital. Data was collected at discharge time and eight weeks post-discharge in the period from April 2019 to December 2021. The SF-12 general health survey and preparedness for caregiving scale were used to measure the caregivers’ QoL and preparedness, respectively. The disability rating scale and the TBI symptom scale were used to measure the patient’s disability and symptoms, respectively. Paired t-test and multiple linear regression analysis were performed. Results: Most caregivers were the parent (41.7%) or child (27.8%) of the patient with TBI. Overall the caregivers had good physical QoL (PQoL) and mental health QoL (MHQoL) but low caregiving preparedness at the time of discharge. At eight weeks post-discharge, there were significant improvements in caregiving preparedness (p< 0.01), patient disability (p< 0.05), and depreciation in caregivers’ MHQoL (p< 0.05), but no change in the PQoL. The modifiable predictors of PQoL were the caregiver’s employment status and the severity of the patient’s sleep and mood problems. The predictors of MHQoL were caregiving preparedness, the patient’s inability to live independently, and the severity of mood and behavioral problems. Conclusion: Omani FCs for patients with TBI experience a negative impact on QoL, and this is correlated with the physical, emotional, and mental health symptoms of the patient. Keywords: Caregiving, Family, Quality of Life, Caregiving Burden, Traumatic Brain Injury, Caregivers’ preparedness, Oman

Independent Prognostic Significance of Monosomy 17 and Impact of Karyotype Complexity in Monosomal Karyotype/Complex Karyotype Acute Myeloid Leukemia: Results from Four ECOG-ACRIN Prospective Therapeutic Trials

The presence of a monosomal karyotype (MK+) and/or a complex karyotype (CK+) identifies subcategories of AML with poor prognosis. The prognostic significance of the most common monosomies (monosomy 5, monosomy 7, and monosomy 17) within MK+/CK+ AML is not well defined. We analyzed data from 1,592 AML patients age 17–93 years enrolled on ECOG-ACRIN therapeutic trials. The majority of MK+ patients (182/195; 93%) were MK+/CK+ with 87% (158/182) having ≥5 clonal abnormalities (CK≥ 5). MK+ patients with karyotype complexity ≤4 had a median overall survival (OS) of 0.4y compared to 1.0y for MK- with complexity ≤4 (p < 0.001), whereas no OS difference was seen in MK+ vs. MK- patients with CK≥ 5 (p = 0.82). Monosomy 5 (93%; 50/54) typically occurred within a highly complex karyotype and had no impact on OS (0.4y; p = 0.95). Monosomy 7 demonstrated no impact on OS in patients with CK≥ 5 (p = 0.39) or CK ≤ 4 (p = 0.44). Monosomy 17 appeared in 43% (68/158) of CK≥ 5 patients and demonstrated statistically significant worse OS (0.4y) compared to CK≥ 5 patients without monosomy 17 (0.5y; p = 0.012). Our data suggest that the prognostic impact of MK+ is limited to those with less complex karyotypes and that monosomy 17 may independently predict for worse survival in patients with AML

'I don't want anyone to know': Experiences of obtaining access to HIV testing by Eastern European, non-European Union sex workers in Amsterdam, the Netherlands.

Historically, the Netherlands has hosted a large number of migrant sex workers. Since sex work is considered a legal profession it might serve as an example of better access to health services, including HIV testing, at least for those working within the legal framework. However, migrant sex workers, especially non-European Union (EU) nationals, might not be eligible to register for official employment and thus face obstacles in obtaining access to health services, becoming essentially invisible. This study examined context-specific vulnerabilities of migrant female sex workers (FSWs) from Belarus, Moldova, Russia and Ukraine, whether and how they have access to HIV testing compared to other EE, non-EU migrant FSWs in Amsterdam in the Netherlands. We conducted a multi-stakeholder perspective study from November 2015 to September 2017 in Amsterdam. The study comprised 1) semi-structured interviews with key stakeholders (N = 19); 2) in-depth interviews with Eastern European, non-EU migrant FSWs (N = 5) and field observations of the escort agency working with them; and 3) in-depth interviews with key stakeholders (N = 12). We found six key barriers to HIV testing: 1) migration and sex-work policies; 2) stigma, including self-stigmatization; 3) lack of trust in healthcare providers or social workers; 4) low levels of Dutch or English languages; 5) negative experience in accessing healthcare services in the home country; and 6) low perceived risk and HIV-related knowledge. Having a family and children, social support and working at the licensed sex-work venues might facilitate HIV testing. However, Internet-based sex workers remain invisible in the sex-work industry. Our findings indicate the importance of addressing women's diverse experiences, shaped by intrapersonal, interpersonal, community, network and policy-level factors, with stigma being at the core. We call for the scaling up of outreach interventions focusing on FSWs and, in particular, migrant FSWs working onlin

'Virus Carriers' and HIV testing: navigating Ukraine's HIV policies and programming for female sex workers

Background: There are an estimated 80,100 female sex workers (FSWs) in Ukraine, of whom 7% are living with HIV. Early HIV diagnosis continues to be a public health priority in Ukraine as only approximately 54% of people living with HIV are diagnosed nationwide. This study aims to analyse the content, context and discourse of HIV testing policies among female sex workers in Ukraine and how these policies are understood and implemented in practice. Methods: To analyse past and current national policies, we searched the database of the Ukrainian Parliament and the Ministry of Health for relevant policy documents (e.g. legislation and orders). To analyse the day-to-day practice of those involved in the implementation of these HIV programmes, we conducted face-to-face semi-structured interviews with key stakeholders. All data were coded using deductive thematic analysis initially guided by the Policy Triangle, a framework which addresses policy content, the process of policy-making, the health policy context, actors involved in policy formulation and implementation. Results: HIV testing policies are formed and implemented in the post-Soviet context through a vertical system of AIDS clinics, resulting in the separation of key affected populations from the rest of the health system. Successive testing policies have been strongly influenced by international donors and non-governmental organisations. Furthermore, a lack of government funding for HIV prevention created a gap that international donors and local non-governmental organisations covered to ensure the implementation of testing policies. Their role, however, had limited influence on the Ukrainian government to increase funding for prevention, including testing of FSWs. Since the early 1990s, when stigmatising and discriminatory forced/mandatory HIV testing was applied, these approaches were slowly replaced with voluntary testing, self-testing and assisted HIV testing, yet stigma was found to be a barrier among FSWs to access testing. Conclusion: Poor governance and the fragmentation of the health system, ongoing health sector reforms, shrinking international funding, and persisting stigma towards people living with HIV and sex workers might impede the continuity and sustainability of HIV testing programmes. Local civil society may now have the opportunity to contribute to the development and further implementation of HIV testing policies in Ukraine

Caregiving Preparedness and Caregiver Burden in Omani Family Caregivers for Patients with Acquired Brain Injury

Objective: To explore the caregiving preparedness and burden among Omani family caregivers (FCs) of patients with acquired brain injury (ABI). Methods: A prospective observational design was used to collect data from 119 FCs and their patients at the time of discharge from the hospital and 16 weeks post-discharge during follow up-care in the neurology clinic. The questionnaire comprised the Zarit Burden Index, the Preparedness for Caregiving Scale, the SF-12 General Health Survey, and a patient symptom scale. Results: FCs were predominantly female (55.5%), and their mean age was 38.27 ± 9.11 years. Most patients had moderate to severe ABI (95.8%) due to stroke (56.3%) and trauma (30.3%). The most common patient symptoms were loss of muscle strength, speech problems, mood problems, memory loss, and change in behavior. Most FCs had a low caregiving preparedness (58%) at discharge, and 19.1% had a high level of caregiving burden at 16 weeks post-discharge. The length of time post-injury (p &lt; 0.01), symptom severity (p &lt; 0.01), and the FCs’ physical and mental health status (p &lt; 0.01) were significant predictors of caregiving preparedness.&nbsp; The predictors of caregiver burden were caregiver preparedness (p &lt; 0.01), symptom severity (p &lt; 0.01), and caregivers’ mental health (p = 0.028). Conclusion: Omani FCs of patients with ABI commence the caregiver role with inadequate preparation, and shortly a significant number suffer a high caregiving burden. Interventions focusing on the caregiver’s health and training in symptom management may enhance the outcomes of FCs and patients. Keywords: Acquired brain injury; Caregiving; Caregiving preparedness; Caregiver burden; Family caregivers; Rehabilitation; Traumatic brain injury; Oman

Interactions between Type 1 Interferons and the Th17 Response in Tuberculosis: Lessons Learned from Autoimmune Diseases

textabstractThe classical paradigm of tuberculosis (TB) immunity, with a central protective role for Th1 responses and IFN-γ-stimulated cellular responses, has been challenged by unsatisfactory results of vaccine strategies aimed at enhancing Th1 immunity. Moreover, preclinical TB models have shown that increasing IFN-γ responses in the lungs is more damaging to the host than to the pathogen. Type 1 interferon signaling and altered Th17 responses have also been associated with active TB, but their functional roles in TB pathogenesis remain to be established. These two host responses have been studied in more detail in autoimmune diseases (AID) and show functional interactions that are of potential interest in TB immunity. In this review, we first identify the role of type 1 interferons and Th17 immunity in TB, followed by an overview of interactions between these responses observed in systemic AID. We discuss (i) the effects of GM-CSF-secreting Th17.1 cells and type 1 interferons on CCR2+ monocytes; (ii) convergence of IL-17 and type 1 interferon signaling on stimulating B-cell activating factor production and the central role of neutrophils in this process; and (iii) synergy between IL-17 and type 1 interferons in the generation and function of tertiary lymphoid structures and the associated follicular helper T-cell responses. Evaluation of these autoimmune-related pathways in TB pathogenesis provides a new perspective on recent developments in TB research