Prevalence of hepatic steatosis in patients with type 2 diabetes and response to glucose-lowering treatments. A multicenter retrospective study in Italian specialist care

Type 2 diabetes (T2D) is a risk factor for metabolic dysfunction-associated fatty liver disease (MAFLD), which is becoming the commonest cause of chronic liver disease worldwide. We estimated MAFLD prevalence among patients with T2D using the hepatic steatosis index (HSI) and validated it against liver ultrasound. We also examined whether glucose-lowering medications (GLM) beneficially affected HSI

Similar effectiveness of dapagliflozin and GLP-1 receptor agonists concerning combined endpoints in routine clinical practice: A multicentre retrospective study

Aims According to cardiovascular outcome trials, some sodium-glucose contransporter-2 inhibitors (SGLT2i) and glucagon-like peptide-1 receptor agonists (GLP-1RA) are recommended for secondary cardiovascular prevention in type 2 diabetes (T2D). In this real-world study, we compared the simultaneous reductions in HbA1c, body weight and systolic blood pressure after initiation of dapagliflozin or GLP-1RA as second or a more advanced line of therapy. Materials and methods DARWIN-T2D was a retrospective multi-centre study conducted at diabetes specialist clinics in Italy that compared T2D patients who initiated dapagliflozin or GLP-1RA (exenatide once weekly or liraglutide). Data were collected at baseline and at the first follow-up visit after 3 to 12 months. The primary endpoint was the proportion of patients achieving a simultaneous reduction in HbA1c, body weight and systolic blood pressure. To reduce confounding, we used multivariable adjustment (MVA) or propensity score matching (PSM). Results Totals of 473 patients initiating dapagliflozin and 336 patients initiating GLP-1RA were included. The two groups differed in age, diabetes duration, HbA1c, weight and concomitant medications. The median follow-up was 6 months in both groups. Using MVA or PSM, the primary endpoint was observed in 30% to 32% of patients, with no difference between groups. Simultaneous reduction of HbA1c, BP and SBP by specific threshold, as well as achievement of final goals, did not differ between groups. GLP-1RA reduced HbA1c by 0.3% more than the reduction achieved with dapagliflozin. Conclusion In routine specialist care, initiation of dapagliflozin can be as effective as initiation of a GLP-1RA for attainment of combined risk factor goals

Colorectal Cancer Stage at Diagnosis Before vs During the COVID-19 Pandemic in Italy

IMPORTANCE Delays in screening programs and the reluctance of patients to seek medical attention because of the outbreak of SARS-CoV-2 could be associated with the risk of more advanced colorectal cancers at diagnosis. OBJECTIVE To evaluate whether the SARS-CoV-2 pandemic was associated with more advanced oncologic stage and change in clinical presentation for patients with colorectal cancer. DESIGN, SETTING, AND PARTICIPANTS This retrospective, multicenter cohort study included all 17 938 adult patients who underwent surgery for colorectal cancer from March 1, 2020, to December 31, 2021 (pandemic period), and from January 1, 2018, to February 29, 2020 (prepandemic period), in 81 participating centers in Italy, including tertiary centers and community hospitals. Follow-up was 30 days from surgery. EXPOSURES Any type of surgical procedure for colorectal cancer, including explorative surgery, palliative procedures, and atypical or segmental resections. MAIN OUTCOMES AND MEASURES The primary outcome was advanced stage of colorectal cancer at diagnosis. Secondary outcomes were distant metastasis, T4 stage, aggressive biology (defined as cancer with at least 1 of the following characteristics: signet ring cells, mucinous tumor, budding, lymphovascular invasion, perineural invasion, and lymphangitis), stenotic lesion, emergency surgery, and palliative surgery. The independent association between the pandemic period and the outcomes was assessed using multivariate random-effects logistic regression, with hospital as the cluster variable. RESULTS A total of 17 938 patients (10 007 men [55.8%]; mean [SD] age, 70.6 [12.2] years) underwent surgery for colorectal cancer: 7796 (43.5%) during the pandemic period and 10 142 (56.5%) during the prepandemic period. Logistic regression indicated that the pandemic period was significantly associated with an increased rate of advanced-stage colorectal cancer (odds ratio [OR], 1.07; 95%CI, 1.01-1.13; P = .03), aggressive biology (OR, 1.32; 95%CI, 1.15-1.53; P < .001), and stenotic lesions (OR, 1.15; 95%CI, 1.01-1.31; P = .03). CONCLUSIONS AND RELEVANCE This cohort study suggests a significant association between the SARS-CoV-2 pandemic and the risk of a more advanced oncologic stage at diagnosis among patients undergoing surgery for colorectal cancer and might indicate a potential reduction of survival for these patients

A Real Time PCR Platform for the Simultaneous Quantification of Total and Extrachromosomal HIV DNA Forms in Blood of HIV-1 Infected Patients

Background: The quantitative measurement of various HIV-1 DNA forms including total, unintegrated and integrated provirus play an increasingly important role in HIV-1 infection monitoring and treatment-related research. We report the development and validation of a SYBR Green real time PCR (TotUFsys platform) for the simultaneous quantification of total and extrachromosomal HIV-1 DNA forms in patients. This innovative technique makes it possible to obtain both measurements in a single PCR run starting from frozen blood employing the same primers and standard curve. Moreover, due to identical amplification efficiency, it allows indirect estimation of integrated level. To specifically detect 2-LTR a qPCR method was also developed. Methodology/Findings: Primers used for total HIV-1 DNA quantification spanning a highly conserved region were selected and found to detect all HIV-1 clades of group M and the unintegrated forms of the same. A total of 195 samples from HIV-1 patients in a wide range of clinical conditions were analyzed with a 100% success rate, even in patients with suppressed plasma viremia, regardless of CD4+ or therapy. No significant correlation was observed between the two current prognostic markers, CD4+ and plasma viremia, while a moderate or high inverse correlation was found between CD4+ and total HIV DNA, with strong values for unintegrated HIV DNA. Conclusions/Significance: Taken together, the results support the use of HIV DNA as another tool, in addition to traditional assays, which can be used to estimate the state of viral infection, the risk of disease progression and to monitor the effects of ART. The TotUFsys platform allowed us to obtain a final result, expressed as the total and unintegrated HIV DNA copy number per microgram of DNA or 10 4 CD4+, for 12 patients within two working day

Post-PCR melt curve analysis and standard curve.

<p>(A) Different dissociation curves that are shown in percentages of their relative amounts in the analysis of 150 HIV-1 negative DNA samples. Only 10 and 2 copies of standard are displayed. (B) Mean standard curve obtained in the <i>TotUFsys</i> PCR experiments (n = 8).</p

Quantification of 2-LTR circles in blood samples.

a<p>Determined subtracting the level of 2-LTR from all unintegrated HIV DNA forms, taking into account the similar PCR efficiencies of 2-LTR and UF.</p>b<p>The median (IQR) value for 2-LTR/µg DNA was 2 (<2–5) and <2 (<2–4) for MDR and non-MDR group, respectively; for 2-LTR/10⁴ CD4+ was 4 (1–7) and 2 (1–3) for the MDR and non-MDR group, respectively.</p><p>The median (IQR) value for UF HIV DNA/µg DNA was 3 (<2–5) and 7 (3–11) for the MDR and non-MDR groups, respectively; for UF HIV DNA/10⁴ CD4+ was 6 (1–9) and 5 (2–10) for the MDR and non-MDR group, respectively.</p><p>The median (IQR) value for 1-LTR + linear forms/µg DNA was 0 (0–2) and 3 (1–8) for the MDR and non-MDR group, respectively; for 1-LTR + linear forms/10⁴ CD4+ was 0 (0–1) and 2 (1–7) for the MDR and non-MDR group, respectively.</p><p>Quantification of 2-LTR circles in blood samples.</p

Correlations between study parameters in blood samples.

<p>Correlations between plasma viremia and CD4+ T cell counts (left panel) and between unintegrated HIV DNA and CD4+ T cell counts (right panel) in (A) a total of 195 blood samples, (B) 85 blood samples collected from patients with multidrug-resistant HIV-1 infection, (C) 110 blood samples collected from patients who had no evidence of resistance, (D) 32 treatment-naïve samples, (E) 163 ART samples, (F) 90 under RAL samples and (G) 72 samples with HIV-1 RNA loads above 50 copies/ml of plasma. Only the correlation between CD4+ and unintegrated HIV DNA is shown because it is stronger than the correlation between CD4+ and total HIV DNA.</p

The effect of the data normalization procedure on the quantification of HIV DNA in blood samples from HIV-1 positive subjects.

<p>The effect of the data normalization procedure on the quantification of HIV DNA in blood samples from HIV-1 positive subjects.</p

Quantification of total and unintegrated (UF) HIV DNA copy number in five different samples.

a<p>The results were obtained dividing the sum of the copy number from a total of eight replicates (two 0.5 µg replicates in the 1^st qPCR and six 0.5 µg replicates in the 2^nd qPCR) by 4 and expressed as HIV DNA copy number/µg of DNA.</p>b<p>Taking into account the WBC number, the results can be reported as copies/ml of blood with the formula: (copies/µg DNA)×WBC no. per ml/142857 cells, assuming that 142857 cells are present in one µg of DNA. WBC counts/µl are 3980 (sample 1), 7480 (sample 2), 8660 (sample 3), 5610 (sample 4), and 6620 (sample 5).</p>c<p>Taking into account %CD4+, the results can be reported as copies/10⁴ CD4+ with the formula: [(copies/µg DNA)/(CD4+/WBC×142857 WBC)]×10⁴. The %CD4+ are: 15.0% (sample 1), 3.2% (sample 2), 12.9% (sample 3), 6.8% (sample 4), and 6.0% (sample 5).</p><p>Quantification of total and unintegrated (UF) HIV DNA copy number in five different samples.</p