Effectiveness and Safety of Postoperative Hospital at Home for Surgical Patients: A Cohort Study.

OBJECTIVE: To determine the feasibility and effectiveness of a Hospital at Home (HaH) enabled early transfer pathways for surgical patients. BACKGROUND: HaH serves as a safe alternative to traditional hospitalization by providing acute care to patients in their homes through a comprehensive range of hospital-level interventions. To our knowledge, no studies have been published to date reporting a large cohort of early home-transferred patients after surgery through a HaH unit. METHODS: Cohort study enrolling every patient admitted to the HaH unit of a tertiary hospital who underwent any of 6 surgeries with a predefined early transfer pathway and fitting both general and surgery inclusion criteria (clinical and hemodynamic stability, uncomplicated surgery, presence of a caregiver, among others) from November 2021 to May 2023. Protocols were developed for each pathway between surgical services and HaH to deliver the usual postoperative care in the home setting. Discharge was decided according to protocol. An urgent escalation pathway was also established. RESULTS: During the study period, 325 patients were included: 141 were bariatric surgeries, 85 kidney transplants, 45 thoracic surgeries, 37 cystectomies, 10 appendicectomies, and 7 ventral hernia repairs. The overall escalation of care during HaH occurred in 7.3% of patients and 30-day readmissions in 7%. Most adverse events were managed at home and the overall mortality was zero. The total mean length of stay was 8 days (interquartile range 2-14), and patients with HaH were transferred home 3 days (interquartile range 1-6) earlier than the usual pathway; a total of 1551 bed-days were saved. CONCLUSIONS: The implementation of early home transfer pathways for surgical patients through HaH is feasible and effective, with favorable safety outcomes.

Optimised molecular genetic diagnostics of Fanconi anaemia by whole exome sequencing and functional studies.

PURPOSE: Patients with Fanconi anaemia (FA), a rare DNA repair genetic disease, exhibit chromosome fragility, bone marrow failure, malformations and cancer susceptibility. FA molecular diagnosis is challenging since FA is caused by point mutations and large deletions in 22 genes following three heritability patterns. To optimise FA patients' characterisation, we developed a simplified but effective methodology based on whole exome sequencing (WES) and functional studies. METHODS: 68 patients with FA were analysed by commercial WES services. Copy number variations were evaluated by sequencing data analysis with RStudio. To test FANCA missense variants, wt FANCA cDNA was cloned and variants were introduced by site-directed mutagenesis. Vectors were then tested for their ability to complement DNA repair defects of a FANCA-KO human cell line generated by TALEN technologies. RESULTS: We identified 93.3% of mutated alleles including large deletions. We determined the pathogenicity of three FANCA missense variants and demonstrated that two FANCA variants reported in mutations databases as 'affecting functions' are SNPs. Deep analysis of sequencing data revealed patients' true mutations, highlighting the importance of functional analysis. In one patient, no pathogenic variant could be identified in any of the 22 known FA genes, and in seven patients, only one deleterious variant could be identified (three patients each with FANCA and FANCD2 and one patient with FANCE mutations) CONCLUSION: WES and proper bioinformatics analysis are sufficient to effectively characterise patients with FA regardless of the rarity of their complementation group, type of mutations, mosaic condition and DNA source.

Clinical Retinal Image Quality of a Non-diffractive Wavefront-Shaping Extended Depth of Focus (Vivity) Intraocular Lens.

PURPOSE: To evaluate clinical retinal optical image quality following implantation of an extended depth of focus intraocular lens (EDOF IOL) (Vivity; Alcon Laboratories, Inc), and to compare it with a monofocal and a trifocal IOL. METHODS: This prospective, comparative, case-control study included 88 eyes implanted with: (1) 19 monofocal IOLs (AcrySof SA60AT; Alcon Laboratories, Inc); (2) 38 EDOF IOLs (AcrySof IQ Vivity); and (3) 31 trifocal IOLs (AT LISA tri 839MP; Carl Zeiss Meditec AG). Total root mean square, ocular lower (LOA) and higher (HOA) order aberrations, point spread function (PSF) Strehl ratio (PSF with LOA), and PSF Strehl ratio excluding LOA (PSF without LOA) were analyzed using a Pyramidal WaveFront-based sensor aberrometer Osiris (Costruzione Strumenti Oftalmici) at two different pupil sizes (3 and 4 mm). RESULTS: The trifocal IOL showed the highest PSF without LOA at both pupil sizes (0.52 ± 0.12 and 0.31 ± 0.07, respectively), followed by the AcrySof SA60AT (0.39 ± 0.10 and 0.27 ± 0.07) and AcrySof IQ Vivity (0.34 ± 0.11 and 0.24 ± 0.09) (P < .001). The AcrySof IQ Vivity and monofocal IOLs were comparable (P > .05). Despite the comparable postoperative low spherical equivalent among the IOL groups, the AT LISA tri 839MP retinal image quality (PSF with LOA) was the most severely affected by such residual refractive errors (dropped to 0.26 ± 0.06 at 3 mm; P < .001) compared to the monofocal AcrySof SA60AT (0.24 ± 0.07 at 3 mm) and EDOF Acrysof IQ Vivity (0.23 ± 0.06 at 3 mm) groups. The PSF with LOA was comparable (P > .05) among the three groups at both the 3-and 4-mm pupil size. CONCLUSIONS: Although trifocal IOLs provided significantly better retinal image quality if influence of LOA is excluded, they also demonstrated to be the most sensitive to residual refractive errors. Both the EDOF Acrysof IQ Vivity and mono-focal AcrySof SA60AT IOLs showed a comparable retinal image quality, and they are also comparable with trifocal IOLs when considering the clinically real PSF (PSF with LOA). [J Refract Surg. 2023;39(2):103-110.].

Modulating mutational outcomes and improving precise gene editing at CRISPR-Cas9-induced breaks by chemical inhibition of end-joining pathways.

Gene editing through repair of CRISPR-Cas9-induced chromosomal breaks offers a means to correct a wide range of genetic defects. Directing repair to produce desirable outcomes by modulating DNA repair pathways holds considerable promise to increase the efficiency of genome engineering. Here, we show that inhibition of non-homologous end joining (NHEJ) or polymerase theta-mediated end joining (TMEJ) can be exploited to alter the mutational outcomes of CRISPR-Cas9. We show robust inhibition of TMEJ activity at CRISPR-Cas9-induced double-strand breaks (DSBs) using ART558, a potent polymerase theta (PolÏ´) inhibitor. Using targeted sequencing, we show that ART558 suppresses the formation of microhomology-driven deletions in favor of NHEJ-specific outcomes. Conversely, NHEJ deficiency triggers the formation of large kb-sized deletions, which we show are the products of mutagenic TMEJ. Finally, we show that combined chemical inhibition of TMEJ and NHEJ increases the efficiency of homology-driven repair (HDR)-mediated precise gene editing. Our work reports a robust strategy to improve the fidelity and safety of genome engineering.

Laser-assisted in situ keratomileusis long term outcomes in late adolescence.

PURPOSE: Evaluate the long term outcomes of myopic-LASIK in a late adolescent population (age ⩾17 and <20 at the time of surgery). METHODS: Monocentric retrospective case series study. Eyes with at least 3 years of follow-up time were included. Primary outcome measures were long term efficacy, safety and stability of the refractive error. Secondary outcome measure was the evaluation of the relation between the postoperative spherical aberration and the long term stability of the refractive error. RESULTS: Forty-seven eyes of 25 patients were included. Mean follow-up was 9.23 ± 3.16 years. Mean age at the time of surgery was 18.74 ± 0.44 years. With time, postoperative UDVA showed a mild but significant deterioration of 1-2 Snellen lines (p = 0.012), in connection with a mild but significant myopization of the SE (mean increase of -0.43 D; p < 0.001), sphere (mean increase of -0.29 D; p = 0.004) and cylinder (mean increase of -0.16 D; p = 0.013). CDVA remained stable over time (p > 0.05). Efficacy index decreased from 1.01 to 0.87 in the long term (77% UDVA ⩾ 20/32). Safety remained at 1.06. 66% and 74% of eyes presented a SE within ±0.50 D and ±1.00 D respectively. SE changed over 0.50D in 33% of eyes. No correlation could be detected between the SE and the postoperative spherical aberration. No cases of corneal ectasia were detected. CONCLUSIONS: Myopic-LASIK in late adolescence is safe and effective, but a mild myopic progression occurs. Despite presence of refractive stability is preferable, if necessary, myopic LASIK provides relatively good outcomes in the long term in this young population.

Small tandem DNA duplications result from CST-guided Pol α-primase action at DNA break termini.

Small tandem duplications of DNA occur frequently in the human genome and are implicated in the aetiology of certain human cancers. Recent studies have suggested that DNA double-strand breaks are causal to this mutational class, but the underlying mechanism remains elusive. Here, we identify a crucial role for DNA polymerase α (Pol α)-primase in tandem duplication formation at breaks having complementary 3' ssDNA protrusions. By including so-called primase deserts in CRISPR/Cas9-induced DNA break configurations, we reveal that fill-in synthesis preferentially starts at the 3' tip, and find this activity to be dependent on 53BP1, and the CTC1-STN1-TEN1 (CST) and Shieldin complexes. This axis generates near-blunt ends specifically at DNA breaks with 3' overhangs, which are subsequently repaired by non-homologous end-joining. Our study provides a mechanistic explanation for a mutational signature abundantly observed in the genomes of species and cancer cells.

Clinical consequences of BRCA2 hypomorphism.

The tumor suppressor FANCD1/BRCA2 is crucial for DNA homologous recombination repair (HRR). BRCA2 biallelic pathogenic variants result in a severe form of Fanconi anemia (FA) syndrome, whereas monoallelic pathogenic variants cause mainly hereditary breast and ovarian cancer predisposition. For decades, the co-occurrence in trans with a clearly pathogenic variant led to assume that the other allele was benign. However, here we show a patient with biallelic BRCA2 (c.1813dup and c.7796 A > G) diagnosed at age 33 with FA after a hypertoxic reaction to chemotherapy during breast cancer treatment. After DNA damage, patient cells displayed intermediate chromosome fragility, reduced survival, cell cycle defects, and significantly decreased RAD51 foci formation. With a newly developed cell-based flow cytometric assay, we measured single BRCA2 allele contributions to HRR, and found that expression of the missense allele in a BRCA2 KO cellular background partially recovered HRR activity. Our data suggest that a hypomorphic BRCA2 allele retaining 37-54% of normal HRR function can prevent FA clinical phenotype, but not the early onset of breast cancer and severe hypersensitivity to chemotherapy.