Hearing impairment in family S1599.

<p><b>A</b> Pedigree of family S1599, indicating the <i>GJB2</i> genotypes of subjects who were analyzed. B Audiograms (air conduction) of affected individuals II:1 and II:6 at ages 50 and 31 years, respectively (circles, right ear; crosses, left ear). The binaural means of the hearing thresholds for air conduction at frequencies 0.5, 1, and 2 kHz for these subjects are 33.3 and 25.8 dB, respectively, corresponding to mild HI (21-40 dB).</p

Sequence of the exon-intron junctions of the single <i>GJB2</i> intron.

<p>Exonic sequence is shown in capitals, whereas intronic sequence is shown in lowercase italics. The three acceptor splice sites identified in this work appear boxed in bold: KS, known acceptor site; AS1, alternative site 1; AS2, alternative site 2. The ATG start codon appears in bold. Nucleotides affected by the c. -22-2A>C and c.35delG mutations are indicated by arrows. Locations of the primers used for identifying <i>GJB2</i> splicing products are underlined.</p

Analysis of <i>GJB2</i> splicing products.

<p>Splicing products from transcription of <i>GJB2</i> were amplified in a fluorescent RT-PCR assay performed with total RNA extracted from saliva samples and were separated by capillary electrophoresis. <b>A </b><i>GJB2</i> splicing products from a normal-hearing control (genotype wt/wt). <b>B </b><i>GJB2</i> splicing products from c. -22-2A>C/c.35delG compound heterozygous subject II:4.</p

Sequence electropherograms of cloned <i>GJB2</i> transcripts from c.

<p><b>-22-2A>C/c.35delG compound heterozygous subject II:4</b>. The stretch of six guanine nucleotides affected by the c.35delG deletion appears boxed in red, while the start ATG codon is boxed in black. <b>A</b> Sequence of the transcript derived from the c.35delG allele, generated by use of the known acceptor site. Note that the stretch of guanine nucleotides contains only five guanines. <b>B</b> Sequence of the transcript derived from the c. -22-2A>C allele generated by use of the alternative acceptor site 1. Note the cytosine nucleotide at c. -22-2, boxed in purple.</p

Relative expression levels of <i>GJB2</i> transcripts generated by splicing at different acceptor sites.

<p>Each panel shows results for a normal-hearing control (genotype wt/wt) and for c. -22-2A>C/c.35delG compound heterozygous subject II:4. <b>A</b> Relative expression of <i>GJB2</i> transcript generated by splicing from the known site (two-tailed <i>p</i> < 0.0001, N=5). B Relative expression of <i>GJB2</i> transcript generated by splicing from alternative site 1 (two-tailed <i>p</i>= 0.0094, N=3).</p

Table_1_Increasing and sustaining blood-borne virus screening in Spain and Portugal throughout the COVID-19 pandemic: a multi-center quality improvement intervention.DOCX

BackgroundAround 57,000 people in Spain and Portugal currently living with HIV or chronic hepatitis C are unaware of their infection. The COVID-19 pandemic severely disrupted screening efforts for these infections. We designed an intervention to increase and sustain opportunistic blood-borne virus (BBV) screening and linkage to care (SLTC) by implementing the TEST model.MethodsThe Plan Do Study Act (PDSA) method of quality improvement (QI) was implemented in 8 healthcare organizations (HCOs), including four hospitals, two clusters of community health centers, and two community-based organizations (CBOs). Baseline assessment included a review of BBV SLTC practices, testing volume, and results 12 months before the intervention. Changes in BBV testing rates over time were measured before, during, and after the COVID-19 lockdowns in 2020. A mixed ANOVA model was used to analyze the possible effect on testing volumes among HCOs over the three study periods.InterventionBBV testing was integrated into normal clinical flow in all HCOs using existing clinical infrastructure and staff. Electronic health record (EHR) systems were modified whenever possible to streamline screening processes, implement systemic institutional policy changes, and promote QI.ResultsTwo years after the launch of the intervention in screening practices, testing volumes increased by 116%, with formal healthcare settings recording larger increases than CBOs. The start of the COVID-19 lockdowns was accompanied by a global 60% decrease in testing in all HCOs. Screening emergency department patients or using EHR systems to automate screening showed the highest resilience and lowest reduction in testing. HCOs recovered 77% of their testing volume once the lockdowns were lifted, with CBOs making the fullest recovery. Globally, enhanced screening techniques enabled HCOs to diagnose a total of 1,860 individuals over the research period.ConclusionsImplementation of the TEST model enabled HCOs to increase and sustain BBV screening, even during COVID-19 lockdowns. Although improvement in screening was noted in all HCOs, additional work is needed to develop strong patient linkage to care models in challenging times, such as global pandemics.</p