Duplex droplet digital PCR to study the composition of HIV-DNA in blood cells

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Minor mutations in HIV protease at baseline and appearance of primary mutation 90M in patients for whom their first protease-inhibitor ntiretroviral regimens failed

The association between minor mutations in human immunodeficiency virus (HIV) protease at baseline and development of common primary mutation 90M at virological failure (conferring some resistance to all protease inhibitors [PIs]) was evaluated in 93 previously drug-naive patients experiencing failure of their first PI-based antiretroviral regimens. In logistic regression analysis, the probability of accumulating a new 90M mutation at virological failure was associated with the presence at baseline of minor mutation 36I (naturally occurring in ∼25% of HIV clade B and in >80% of HIV non-clade-B viruses) (adjusted odds ratio, 13.5 [95% confidence interval, 1.89–95.6]; P=.009) and, possibly, of 10I/V. This suggests a potential role for the presence of 36I at baseline in predicting the appearance of 90M at virological failure

Contribution of transgender sex workers to the complexity of the HIV-1 epidemic in the metropolitan area of Milan

Objectives: Transgender people are disproportionately affected by the HIV-1 epidemic. We evaluated the origin of HIV-1 variants carried by South American transgenders living in Milan by combining accurate phylogenetic methods and epidemiological data. Methods: We collected 156 HIV-1 pol sequences obtained from transgender patients engaged in sex work (TSWs) followed between 1999 and 2015 at L. Sacco Hospital, Milan, Italy. Phylogenetic analyses were conducted by HIV-TRACE, MrBayes, MacClade and Beast programs. Reference sequences were retrieved from Los Alamos and local databases. Last negative testing or proxy data from clinical records of infected individuals were used to investigate the country of infection. Results: Among South American TSWs, the most represented HIV-1 subtypes were B (70.5%), F1 (12.8%) and C (4.4%). Gene flow migrations of B subtype indicated significant fluxes from TSWs to Italians (21.3%) belonging to all risk groups (26.4% to heterosexuals (HEs), 18.9% to men who have sex with men (MSM), 15.1% to injecting drug users). The largest proportion of bidirectional fluxes were observed between Italians and TSWs (24.6%). For F1 subtype, bidirectional viral fluxes involved TSWs and Italians (7.1% and 14.3%), and a similar proportion of fluxes linked TSWs and Italian HEs or MSM (both 15.8%). Significant fluxes were detected from Italians to TSWs for subtype C involving both MSM (30%) and HEs (40%). Country of HIV-1 acquisition was identified for 72 subjects; overall, the largest proportion of patients with B subtype (73.5%) acquired HIV-1 infection in South America. Conclusions: Our results indicated that South American transgenders largely contribute to the heterogeneity of HIV-1 variants in our country. The high number of clusters based on all subtypes indicated numerous transmission chains in which TSWs were constantly intermixed with HEs and MSM. Our results strongly advocate interventions to facilitate prevention, diagnosis and HIV-1 care continuum among transgender people

Immunovirological Response to Triple Nucleotide Reverse-Transcriptase Inhibitors and Ritonavir-Boosted Protease Inhibitors in Treatment-Naive HIV-2-Infected Patients: The ACHIEV2E Collaboration Study Group

This European inter-cohort collaboration shows poorer immunological and virological responses with triple NRTI compared to PI/r-containing cART in treatment-naïve HIV-2-infected patients, regardless of baseline CD4 cell count. PI/r-containing cART should be recommended as first-line antiretroviral regimens in HIV-2 infectio

Wound Myiasis Caused by Sarcophaga (Liopygia) Argyrostoma

In Mediterranean countries, Sarcophaga (Liopygia) crassipalpis, Sarcophaga (L.) argyrostoma, and Sarcophaga (L.) cultellata share the same ecological niche and can be responsible of myiasis. In this study, the main morphological characters of a larva found in a hospitalized woman were described and illustrated by light and SEM microscopy and the features discussed. Then, a fragment within the mitochondrial encoded cytochrome c oxidase subunit I (coxI) gene of ~735 bp was amplified and sequenced. The molecular investigation was necessary to confirm the species Sarcophaga (Liopygia) argyrostoma (99% of identity). Our findings showed that morphological descriptions of larvae of three Mediterranean species of Liopygia available in several papers might not be clear enough to allow for comparison and correct identification. Until results of reliable comparative studies of larvae of all three species will be available, the use of molecular tools is crucial, to avoid misleading or incomplete identification, and in particular when a myiasis becomes a legal issue

Local epidemics gone viral: Evolution and diffusion of the Italian HIV-1 recombinant form CRF60_BC

The molecular epidemiology of HIV-1 in Italy is becoming increasingly complex, mainly due to the spread of non-B subtypes and the emergence of new recombinant forms. We previously characterized the outbreak of the first Italian circulating recombinant form (CRF60_BC), occurring among young MSM living in Apulia between the years 2009 and 2011. Here we show a 5-year follow-up surveillance to trace the evolution of CRF60_BC and to investigate its further spread in Italy. We collected additional sequences and clinical data from patients harboring CRF60_BC, enrolled at the Infectious Diseases Clinic of the University of Bari. In addition to the 24 previously identified sequences, we retrieved 27 CRF60_BC sequences from patients residing in Apulia, whose epidemiological and clinical features did not differ from those of the initial outbreak, i.e., the Italian origin, young age at HIV diagnosis (median: 24 years; range: 18-37), MSM risk factor (23/25, 92%) and recent infection (from 2008 to 2017). Sequence analysis revealed a growing overall nucleotide diversity, with few nucleotide changes that were fixed over time. Twenty-seven additional sequences were detected across Italy, spanning multiple distant regions. Using a BLAST search, we also identified a CRF60_BC sequence isolated in United Kingdom in 2013. Three patients harbored a unique second generation recombinant form in which CRF60_BC was one of the parental strains. Our data show that CRF60_BC gained epidemic importance, spreading among young MSM in multiple Italian regions and increasing its population size in few years, as the number of sequences identified so far has triplicated since our first report. The observed further divergence of CRF60_BC is likely due to evolutionary bottlenecks and host adaptation during transmission chains. Of note, we detected three second-generation recombinants, further supporting a widespread circulation of CRF60_BC and the increasing complexity of the HIV-1 epidemic in Italy

Changing patterns in HIV-1 non-B clade prevalence and diversity in Italy over three decades

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WEED CONTROL AND SELECTIVITY OF SULFENTRAZONE + DIURON ON SUGAR CANE (CULTIVAR RB 966928)

A seletividade e eficiência são características fundamentais para a recomendação de herbicidas. Objetivou-se com este trabalho avaliar o controle de plantas daninhas e a seletividade em cana-de-açúcar da mistura comercial sulfentrazone + diuron (Stone, 700 g i.a. ha-1 de sulfentrazone + 1400 g i.a. ha-1 de diuron, SC, FMC) em cinco doses: D1: zero g i.a. ha-1; D2: 350 g i.a. ha-1 + 700 g i.a. ha-1; D3: 700 g i.a. ha-1 + 1400 g i.a. ha-1; D4: 1050 g i.a. ha-1 + 2100 g i.a. ha-1; D5: 1400 g i.a. ha-1 + 2800 g i.a. ha-1; e em duas modalidades de aplicação: pré e pósemergência inicial. Foram conduzidos dois experimentos, um para cada modalidade de aplicação, em delineamento inteiramente casualisado, com quatro repetições. Foi avaliado o controle de plantas daninhas e a eficiência fotoquímica máxima do fotossistema II (Fv/Fm), a estatura, massa fresca e massa seca da parte aérea e o volume e massa seca de raízes da cana-de-açúcar. A aplicação em pósemergência da mistura comercial sulfentrazone + diuron afetou o Fv/Fm em todas as doses avaliadas. Porém, as menores doses do herbicida (D2 e D3) não influenciaram negativamente nas variáveis fitométricas. Mesmo na menor dose (D2), o controle das plantas daninhas foi total durante o período de avaliações. As doses de sulfentrazone + diuron, entre 350 g i.a. ha-1 + 700 g i.a. ha-1 e 700 g i.a. ha-1 + 1400 g i.a. ha-1, em pré e pós-emergência inicial, são eficientes no controle de plantas daninhas e seletivas para a cana-de-açúcar cultivar RB 966928.Selectivity and efficiency are key characteristics for herbicide recommendation. The objective of this work was to evaluate weed control and sugarcane selectivity of the commercial sulfentrazone + diuron mixture (Stone, 700 g a.i. ha-1 of sulfentrazone + 1400 g a.i. ha-1 of diuron, SC, FMC) in five doses: D1: zero g a.i. ha-1; D2: 350 g a.i. ha-1 + 700 g a.i. ha-1; D3: 700 g a.i. ha-1 + 1400 g a.i. ha-1; D4: 1050 g a.i. ha-1 + 2100 g a.i. ha-1; D5: 1400 g a.i. ha-1 + 2800 g a.i. ha-1; in two application modalities: pre- and initial post-emergence. Two experiments were conducted, one for each application mode, with four replications in completely randomized design. Weed control, maximum photochemical efficiency of photosystem II (Fv/Fm), height, fresh mass and dry mass of shoot and root volume and dry mass of sugarcane were evaluated. Post-emergence application of the commercial sulfentrazone + diuron mixture affected Fv/Fm at all doses evaluated. However, the lower herbicide doses (D2 and D3) did not negatively influence the phytometric variables. Even at the lowest dose (D2) weed control was total during the evaluation period. Sulfentrazone + diuron doses, between 350 g a.i. ha-1 + 700 g a.i. ha-1 and 700 g a.i. ha-1 + 1400 g a.i. ha-1, in pre- and initial post-emergence, are efficient in weed control and selective for sugarcane cultivar RB 966928

Molecular and phylogenetic analysis of HIV-1 variants circulating in Italy

<p>Abstract</p> <p>Objective</p> <p>The continuous identification of HIV-1 non-B subtypes and recombinant forms in Italy indicates the need of constant molecular epidemiology survey of genetic forms circulating and transmitted in the resident population.</p> <p>Methods</p> <p>The distribution of HIV-1 subtypes has been evaluated in 25 seropositive individuals residing in Italy, most of whom were infected through a sexual route during the 1995–2005 period. Each sample has been characterized by detailed molecular and phylogenetic analyses.</p> <p>Results</p> <p>18 of the 25 samples were positive at HIV-1 PCR amplification. Three samples showed a nucleotide divergence compatible with a non-B subtype classification. The phylogenetic analysis, performed on both HIV-1 <it>env </it>and <it>gag </it>regions, confirms the molecular sub-typing prediction, given that 1 sample falls into the C subtype and 2 into the G subtype. The B subtype isolates show high levels of <it>intra</it>-subtype nucleotide divergence, compatible with a long-lasting epidemic and a progressive HIV-1 molecular diversification.</p> <p>Conclusion</p> <p>The Italian HIV-1 epidemic is still mostly attributable to the B subtype, regardless the transmission route, which shows an increasing nucleotide heterogeneity. Heterosexual transmission and the interracial blending, however, are slowly introducing novel HIV-1 subtypes. Therefore, a molecular monitoring is needed to follow the constant evolution of the HIV-1 epidemic.</p