The Anopheles gambiae glutathione transferase supergene family: annotation, phylogeny and expression profiles

BACKGROUND: Twenty-eight genes putatively encoding cytosolic glutathione transferases have been identified in the Anopheles gambiae genome. We manually annotated these genes and then confirmed the annotation by sequencing of A. gambiae cDNAs. Phylogenetic analysis with the 37 putative GST genes from Drosophila and representative GSTs from other taxa was undertaken to develop a nomenclature for insect GSTs. The epsilon class of insect GSTs has previously been implicated in conferring insecticide resistance in several insect species. We compared the expression level of all members of this GST class in two strains of A. gambiae to determine whether epsilon GST expression is correlated with insecticide resistance status. RESULTS: Two A. gambiae GSTs are alternatively spliced resulting in a maximum number of 32 transcripts encoding cytosolic GSTs. We detected cDNAs for 31 of these in adult mosquitoes. There are at least six different classes of GSTs in insects but 20 of the A. gambiae GSTs belong to the two insect specific classes, delta and epsilon. Members of these two GST classes are clustered on chromosome arms 2L and 3R respectively. Two members of the GST supergene family are intronless. Amongst the remainder, there are 13 unique introns positions but within the epsilon and delta class, there is considerable conservation of intron positions. Five of the eight epsilon GSTs are overexpressed in a DDT resistant strain of A. gambiae. CONCLUSIONS: The GST supergene family in A. gambiae is extensive and regulation of transcription of these genes is complex. Expression profiling of the epsilon class supports earlier predictions that this class is important in conferring insecticide resistance

Occupational Therapy for Parkinsonian Patients: A Retrospective Study

Background: Hand functionality and finger dexterity are impaired in patients with Parkinson’s disease (PD). These disturbances lead to a dependency in activities of daily living (ADL) and poor quality of life (QoL). Objective: We aimed to evaluate whether a specific occupational therapy (OT) program is effective in improving finger and hand dexterity and its impact on ADL in PD patients. Methods: We retrospectively studied PD patients, hospitalized for a 4-week multidisciplinary intensive rehabilitation treatment (MIRT) between January 2015 and June 2018. All patients underwent 1 h/day OT treatment, 5 days a week. The primary outcome measure was the O’Connor finger dexterity test; secondary outcome measures were the Minnesota dexterity test, UPDRS II, and Self-Assessment Parkinson’s Disease Disability Scale (SPDDS). These measures were assessed at admission (T0) and discharge (T1). Results: Based on the Hoehn and Yahr scale (H&Y), patients were divided into two groups: 262 subjects in H&Y stage <3 (early-stage PD patients) and 220 in H&Y stage ≥3 (medium-advanced stage PD patients). As expected, at baseline, all measures were worse in higher H&Y stages. After treatment, both groups experienced significant improvements in all outcomes. Significant differences between early-stage and medium-advanced stage PD patients were observed only for the changes in UPDRS II, with a better improvement in patients in H&Y stage ≥3. Conclusions: We showed that PD patients who underwent a rehabilitation protocol including OT experienced improvements in finger dexterity and hand functionality. Our results underline the relevance of OT in improving autonomy and QoL in PD patients

Antibody Complementarity-Determining Regions (CDRs): A Bridge between Adaptive and Innate Immunity

Background: It has been documented that, independently from the specificity of the native antibody (Ab) for a given antigen (Ag), complementarity determining regions (CDR)-related peptides may display differential antimicrobial, antiviral and antitumor activities. Methodology/Principal Findings: In this study we demonstrate that a synthetic peptide with sequence identical to VHCDR3 of a mouse monoclonal Ab (mAb) specific for difucosyl human blood group A is easily taken up by macrophages with subsequent stimulation of: i) proinflammatory cytokine production; ii) PI3K-Akt pathway and iii) TLR-4 expression. Significantly, V HCDR3 exerts therapeutic effect against systemic candidiasis without possessing direct candidacidal properties. Conclusions/Significance: These results open a new scenario about the possibility that, beyond the half life of immunoglobulins, Ab fragments may effectively influence the antiinfective cellular immune response in a way reminiscen

Beating Cancer by 2030: Mission Impossible?

Cancer has been for many years the second leading cause of mortality right after cardiovascular diseases, representing 25% of all the deaths reported yearly and this tendency is expected to increase. Although the recent public health emergency caused by COVID-19 pandemic diverted much of the attention of policy makers, the public opinion and even researchers from other important, economical relevant and deadly diseases, cancer still remains as one of the major healthcare issues. Moreover, recent studies revealed the negative effects of COVID-19 pandemic on the increase of avoidable cancer-related deaths. It is then the perfect time to bring back the spotlight onto the topic of cancer.The aim of this paper is to share the outcomes of the workshop organized by the COST (European Cooperation in Science and Technology) Association, bringing together sixty participants representing a broad variety of stakeholders, to discuss a holistic approach on how to beat cancer by 2030.The conclusions of this workshop are highly relevant for the community and are supporting the work being undertaken by the EU Mission Board on Cancer. This report lays down the main conclusions and recommendations agreed by the workshop participants, focusing on different aspects such as better stakeholder collaboration, citizen education, innovative therapies, and patient-centric care

Heterologous expression of four glutathione transferase genes genetically linked to a major insecticide-resistance locus from the malaria vector Anopheles gambiae.

A cluster of eight genes encoding glutathione transferases (GSTs) are located on division 33B of polytene chromosome arm 3R of the African malaria mosquito, Anopheles gambiae. This region of the genome contains a major 1,1,1-trichloro-2,2-bis-( p -chlorophenyl)ethane (DDT)-resistance locus, rtd1. These GSTs belong to the insect-specific Epsilon class and share between 22.6 and 65.2% identity at the amino acid level. Two distinct allelic variants of the Epsilon GST, GSTe1, differing at 12 out of 224 amino acid residues, are present in laboratory and field populations of A. gambiae. To investigate the possible role of these GSTs in conferring resistance to the insecticide DDT, both GSTe1 alleles, plus three additional members of this gene cluster, were expressed in Escherichia coli and the recombinant proteins biochemically characterized. The five putative glutathione transferases encoded catalytically active subunits with variable biochemical properties. For example, the two allelic variants of GSTE1-1 encoded proteins with over 100-fold variation in peroxidase activity, while the three remaining GSTs had no detectable peroxidase activity. Only GSTE2-2 was able to metabolize DDT. Western blots using antibodies raised against these GSTs indicated that the expression of GSTE2-2 is elevated in a DDT-resistant strain of A. gambiae

Occupational Therapy for Parkinsonian Patients: A Retrospective Study

Background: Hand functionality and finger dexterity are impaired in patients with Parkinson’s disease (PD). These disturbances lead to a dependency in activities of daily living (ADL) and poor quality of life (QoL). Objective: We aimed to evaluate whether a specific occupational therapy (OT) program is effective in improving finger and hand dexterity and its impact on ADL in PD patients. Methods: We retrospectively studied PD patients, hospitalized for a 4-week multidisciplinary intensive rehabilitation treatment (MIRT) between January 2015 and June 2018. All patients underwent 1 h/day OT treatment, 5 days a week. The primary outcome measure was the O’Connor finger dexterity test; secondary outcome measures were the Minnesota dexterity test, UPDRS II, and Self-Assessment Parkinson’s Disease Disability Scale (SPDDS). These measures were assessed at admission (T0) and discharge (T1). Results: Based on the Hoehn and Yahr scale (H&Y), patients were divided into two groups: 262 subjects in H&Y stage <3 (early-stage PD patients) and 220 in H&Y stage ≥3 (medium-advanced stage PD patients). As expected, at baseline, all measures were worse in higher H&Y stages. After treatment, both groups experienced significant improvements in all outcomes. Significant differences between early-stage and medium-advanced stage PD patients were observed only for the changes in UPDRS II, with a better improvement in patients in H&Y stage ≥3. Conclusions: We showed that PD patients who underwent a rehabilitation protocol including OT experienced improvements in finger dexterity and hand functionality. Our results underline the relevance of OT in improving autonomy and QoL in PD patients

Phospho-IkBα activation and TNF-α gene expression in PM stimulated with V_HCDR3.

<p>PM (3×10⁶/ml) were stimulated for 1 h in the presence or absence (NS) of wortmannin (4 nM), V_HCDR3, LPS or NC (all at 10 µg/ml). After incubation, cell lysates were subjected to Western blotting. Membranes were incubated with Abs to pIkBα and IkBα; pIkBα was normalized against IkBα. (<b>A</b>) *, <i>P</i><0.05 (treated <i>vs</i> untreated cells, n = 5); †, <i>P</i><0.05 (wortmannin-treated <i>vs</i> wortmannin-untreated cells, n = 5). For testing the expression level of TNF-α gene, PM (1×10⁶/ml) were cultured for 1, 6 and 18 h as above described. After incubation, total RNA was isolated and analyzed for mRNA expression with RT-PCR. Transcript copy numbers were determined by qPCR using cDNA as a template. Copy numbers were normalized against the copy number of the GADPH gene (<b>B</b>). *, <i>P</i><0.05 (treated <i>vs</i> untreated cells, n = 5). Error bars, s.e.m.</p

Primer sequences used for quantitative RT-PCR.

<p>Primers were designed with the help of Beacon Designer software (Bio-Rad, Hercules, CA, USA) and provided by Invitrogen.</p