Clinical Features, Cardiovascular Risk Profile, and Therapeutic Trajectories of Patients with Type 2 Diabetes Candidate for Oral Semaglutide Therapy in the Italian Specialist Care

Introduction: This study aimed to address therapeutic inertia in the management of type 2 diabetes (T2D) by investigating the potential of early treatment with oral semaglutide. Methods: A cross-sectional survey was conducted between October 2021 and April 2022 among specialists treating individuals with T2D. A scientific committee designed a data collection form covering demographics, cardiovascular risk, glucose control metrics, ongoing therapies, and physician judgments on treatment appropriateness. Participants completed anonymous patient questionnaires reflecting routine clinical encounters. The preferred therapeutic regimen for each patient was also identified. Results: The analysis was conducted on 4449 patients initiating oral semaglutide. The population had a relatively short disease duration (42%  60% of patients, and more often than sitagliptin or empagliflozin. Conclusion: The study supports the potential of early implementation of oral semaglutide as a strategy to overcome therapeutic inertia and enhance T2D management

In-field and in-vitro study of the moss Leptodictyum riparium as bioindicator of toxic metal pollution in the aquatic environment: ultrastructural damage, oxidative stress and HSP70 induction.

This study evaluate the effects of freshwater toxic metal pollution in the highly contaminated Sarno River (South Italy), by using the aquatic moss L. riparium in bags at 3 representative sites of the river. Biological damage was assessed by studying metal bioaccumulation, ultrastructural changes, oxidative stress, as Reactive Oxygen Species (ROS) production and Glutathione S-transferase (GST ) activity, and Heat Shock Proteins 70 (HSP70) induction. The results showed that L. riparium is a valuable bioindicator for toxic metal pollution of water ecosystem, accumulating different quantities of toxic metals from the aquatic environment. Toxic metal pollution caused severe ultrastructural damages, such as the increase of ROS production and the induction of GST and HSP70s in the samples from the polluted sites. To assess the role and the effects of toxic metals on plants, L. riparium samples cultured in vitro were exposed to Cd, Cr, Cu, Fe, Ni, Pb, Zn at the same concentrations as measured at the 3 sites. Ultrastructural damages, ROS, GST, HSP70s were also severely affected by toxic metals. From our finding we can conclude that L. riparium can be proposed as a model organism in biomonitoring projects, and GST and HSP70s as promising biomarkers of metal toxicity

In-field and in-vitro study of the moss <i>Leptodictyum riparium</i> as bioindicator of toxic metal pollution in the aquatic environment: Ultrastructural damage, oxidative stress and HSP70 induction - Fig 1

<p><b>TEM micrographs from leaflets of <i>L</i>. <i>riparium</i> specimens exposed in the river Sarno at the site A (1–5), site B (6–10) and site C (11–15).</b> Site A. (1) Thick wall delimited cells showing lenticular chloroplasts, with grana and starch grains, and large clear vacuoles occupying the centre of the protoplast. (2) A thick walled cell with regular chloroplasts and vacuole. The chloroplasts show well-developed grana. (3) The chloroplasts contain a well-developed thylakoid system, starch grains and rare plastoglobules. (4) The thick wall delimited cell shows regular chloroplasts, with grana and starch grains, and a central nucleus, with eu- and heterochromatin. (5) A section of a mitochondrion with cristae. Site B. (6) A thick wall delimited cell containing chloroplasts, with grana and plastoglobules, and cytoplasm lipid droplets. (7) The cell contains a miss-shaped chloroplast with grana and plastoglobules, cytoplasm lipid droplets and vesicles. (8) A miss-shaped chloroplast with a well-developed thylakoid system and plastoglobules. Lipid droplets and a mitochondrion with no cristae are between the chloroplasts. (9) A chloroplast with a well-developed thylakoid system and plastoglobules. (10) Vesicles at high magnification. Site C. (11) A severely altered cell featured by a highly fissured thick wall. The chloroplasts, still showing a developed thylakoid system with grana, are swollen and filled with large plastoglobules. Large lipid droplets are in the cytoplasm. (12) The altered cell shows cytoplasm lipid droplets and a swollen chloroplast with plastoglobules and thylakoids. (13–14) Chloroplasts showing large plastoglobules and thylakoid systems with still recognizable grana and intergrana membranes. Large lipid droplets around the chloroplast. (15) Magnified plastoglobules and thylakoids. Scala bars: 5 μ (1), 2 μ (4), 1 μ (2, 6, 7, 11, 15), 500 nm (3, 9, 12, 13, 14), 300 nm (5, 10)<b>. Lettering and marks: cw</b> cell wall; <b>m</b> mitochondrion; <b>n</b> nucleus; <b>*</b> starch grain; <b>black arrow</b> cytoplasm lipid droplet; <b>white arrow</b> plastoglobules.</p

In-field and in-vitro study of the moss <i>Leptodictyum riparium</i> as bioindicator of toxic metal pollution in the aquatic environment: Ultrastructural damage, oxidative stress and HSP70 induction - Fig 2

<p>The table shows TEM micrographs from leaflets of <i>L</i>. <i>riparium</i> specimens cultured in the toxic metal mixture at the same concentrations as in the site A (1–5), site B (6–10) and site C (11–15). Site A. (1) Thick wall delimited cells containing lenticular chloroplasts, with grana and starch grains, and large clear vacuoles. (2, 3) Regular chloroplasts featured by a well-developed thylakoid system and mitochondria. (4) A central nucleus (N) with eu- and heterochromatin, surrounded by chloroplasts and vacuoles. (5) A section of a mitochondrion with cristae. Site B. (6) Thick wall delimited cells showing plasmolysed protoplasts with severely swollen chloroplasts. (7, 8) Severely plasmolysed cells containing swollen chloroplasts with swollen thylakoids. (9, 10) Swollen chloroplasts with swollen thylakoids and small starch grains. Membranes have a thicker and not sharp appearance. Site C. (11) Inside the thick wall delimited cells are changed chloroplasts, vacuoles and a nucleus. (12) A cell with chloroplasts and plenty of cytoplasm vesicles. (13) A detail of a cell showing a misshaped chloroplast with grana and large plastoglobules in the stroma. A nucleus with eu- and etherochromatin is on the left. (14) Details of a misshaped chloroplast with grana, mitochondria and a multilamellar body. (15) A section of a mitochondrion with cristae. Scale bars: 5 μm (1), 3 μm (11), 2 μm (6, 12), 1 μm (4, 7, 8), 500 nm (2, 3, 9, 10, 13, 14), 300 nm (5, 15). Lettering and marks: cw cell wall; m mitochondrion; n nucleus; v vacuole; * starch grain; white arrow plastoglobules; + multilamellar body.</p

GST activity in <i>L</i>. <i>riparium</i> exposed in bags and <i>in vitro</i> cultured.

<p>GST activity in <i>L</i>. <i>riparium</i> exposed in bags at sites A, B and C of Sarno River (in-field experiment, left panel, a) and <i>in vitro</i> cultured with mixtures of toxic metal or with the single toxic metal at the concentrations measured in site C (CdCl₂ 0.14 mg l^-1, Cr(Cl) 3 9.05 mg l^-1, CuSO₄ 2.45 mg l^-1, FeCl₂ 308.0 mg l^-1, NiCl₂ 3.4 mg l^-1, Pb(CH₃COO)₂ 0.85 mg l^-1, ZnCl₂ 46.76 mg l^-1) (right panel, b). Data are shown as the mean ± standard deviation (n = 5). The GST activity was expressed as micromoles/ml min^-1. Bars not accompanied by the same letter are significantly different at p < 0.05, using post hoc Student-Neuman-Keuls test.</p

The table shows TEM micrographs from <i>L</i>. <i>riparium</i> leaflets of samples treated with the single toxic metals.

<p><b>(1–5) Cd-treated samples.</b> (1) Thick wall delimited cells with chloroplasts, nucleus and vacuoles. (2) A plasmolysed cell showing a large vacuole and chloroplasts with grana. (3) A misshaped chloroplast with grana and integrana thylakoids. (4) A bulge from a misshaped chloroplast. (5) Beside a chloroplast mitochondria with cristae. <b>(6–10) Cr-treated samples.</b> (6) Thick wall delimited cells with highly vacuolated cytoplasm, chloroplasts and nuclei. (7) The cell cytoplasm contains clear and electron dense vacuoles, chloroplasts and a nucleus. (8) The plasmolysed cell presents a misshaped chloroplast, with grana and a starch grain, clear and electron dense vacuoles, a nucleus and a large lipid droplet in the cytoplasm. (9) The cell contains a central nucleus surrounded by misshaped chloroplasts with a poorly developed thylakoid system and visible plastoglobules. (10) A diving mitochondrion, with clear matrix and no cristae, located beside a cytoplasm lipid droplet. <b>(11–15) Cu-treated sample.</b> (11–12) Severely plasmolysed cells with swollen chloroplasts and empty cells are shown. (13) A plasmolysed cell filled with swollen chloroplasts provided by starch grains. (14–15) Plasmolysed cells containing swollen chloroplasts with visible grana and starch grains. A multilamellar body near the plasma membrane. All the membranes appear poorly sharp. <b>(16–20) Fe-treated samples.</b> (16–18) Thick wall delimited cells with misshaped chloroplasts containing grana. (19) A normal nucleus with eu- and heterochromatin and a nucleolus. (20) A section of a mitochondrion with cristae. <b>(21–25) Ni-treated samples.</b> (21) The thick wall delimited cells show chloroplasts with grana and clear vacuoles. (22–23) Cells containing clear vacuoles and chloroplasts with grana. (24) A nucleus next to a chloroplast. (25) A section of a mitochondrion with developed cristae. <b>(26–30) Pb-treated samples.</b> (26) Thick wall delimited cells with chloroplasts and large clear vacuoles. (27) A cell contains chloroplasts with grana and clear vacuoles. Some of the chloroplasts appear misshaped. (28) A misshaped chloroplast with grana. (29) A vesicle filled with material fuses with plasma membrane. (30) A longitudinal section of a mitochondrion with cristae <b>(31–35) Zn-treated samples.</b> (31) Cells with chloroplasts containing starch grains. (32) A chloroplast with grana and starch grains. (33) Plenty of vesicles in the cytoplasm beside dictyosomes and chloroplasts. (34) A multilamellar body. (35) A longitudinal section of a mitochondrion with cristae, next to cytoplasm vesicles. <b>Scale bars: 5</b> μ<b>m</b> (1, 6, 11, 16, 21, 26, 31), <b>2</b> μ<b>m</b> (2, 7, 12, 17, 22, 27), <b>1</b> μ<b>m</b> (3, 8, 9, 13, 18, 19, 23, 24, 28, 32, 33), <b>500 nm</b> (4, 14, 15, 29, 30), <b>300 nm</b> (5, 10, 20, 25, 34, 35)<b>. Lettering and marks: chl</b> chloroplast; <b>cw</b> cell wall; <b>d</b> dictyosomes; <b>m</b> mitochondrion; <b>n</b> nucleus; <b>v</b> vacuole; <b>ve</b> vesicles; <b>*</b> starch grain; <b>white arrow</b> plastoglobules; <b>+</b> multilamellar body.</p

In-field and in-vitro study of the moss <i>Leptodictyum riparium</i> as bioindicator of toxic metal pollution in the aquatic environment: Ultrastructural damage, oxidative stress and HSP70 induction - Fig 6

<p><b>Western blotting using Hsp70 antibodies (Sigma) of <i>L</i>.<i>riparium</i> samples exposed along the Sarno River at sites A, B and C.</b> The lower figure shows Western Blotting of the same samples using alfa-tubulin to check the equal loading of electrophoretic lanes.</p

Western blotting using Hsp70 antibodies (Sigma) of <i>L</i>. <i>riparium</i> samples exposed <i>in vitro</i>.

<p>Western blotting using Hsp70 antibodies (Sigma) of <i>L</i>. <i>riparium</i> samples exposed <i>in vitro</i> to metal mixtures of site C and site A, and to the single metal concentrations as measured in the site C: CdCl₂ 0.14 mg l^-1, Cr(Cl) 3 9.05 mg l^-1, CuSO₄ 2.45 mg l^-1, FeCl₂ 308.0 mg l^-1, NiCl₂ 3.4 mg l^-1, Pb(CH₃COO)₂ 0.85 mg l^-1, ZnCl₂ 46.76 mg l^-1.</p

Concentrations (μg g^-1 dw) of toxic metals in moss bags of <i>L</i>. <i>riparium</i> exposed for 7 days at the 3 sites (A, B, C) along the river Sarno.

<p>Concentrations (μg g^-1 dw) of toxic metals in moss bags of <i>L</i>. <i>riparium</i> exposed for 7 days at the 3 sites (A, B, C) along the river Sarno.</p

In-field and in-vitro study of the moss <i>Leptodictyum riparium</i> as bioindicator of toxic metal pollution in the aquatic environment: Ultrastructural damage, oxidative stress and HSP70 induction - Fig 4

<p><b>ROS content in <i>L</i>. <i>riparium</i> exposed in bags at sites A, B and C of Sarno River (in-field experiment, left panel, a) and <i>in vitro</i> cultured with mixtures of toxic metal or with the single toxic metal at the concentrations measured in site C (CdCl</b>_<b>2</b><b>0.14 mg l</b>^<b>-1</b><b>, Cr(Cl) 3 9.05 mg l</b>^<b>-1</b><b>, CuSO</b>_<b>4</b><b>2.45 mg l</b>^<b>-1</b><b>, FeCl</b>_<b>2</b><b>308.0 mg l</b>^<b>-1</b><b>, NiCl</b>_<b>2</b><b>3.4 mg l</b>^<b>-1</b><b>, Pb(CH3COO)</b>_<b>2</b><b>0.85 mg l</b>^<b>-1</b><b>, ZnCl</b>_<b>2</b><b>46.76 mg l</b>^<b>-1</b><b>) (right panel, b).</b> Data are shown as the mean ± standard deviation (n = 5). The ROS quantity was monitored by fluorescence (excitation wavelength of 350 nm and emission wavelength of 600 nm). Bars not accompanied by the same letter are significantly different at p < 0.05, using post hoc Student-Neuman-Keuls test.</p