Elucidating metformin action on the ovary and fertility in healthy mice.

In brief: The hypoglycemic drug metformin has shown reproductive effects in women, although its mechanism of action is not fully understood. In this study, we demonstrate the direct effects of metformin on the ovary of healthy mice, with no alterations in fertility. Abstract: Metformin is a hypoglycemic drug widely used in type-2 diabetes (T2D) patients. In recent years, this drug has been suggested as a treatment for gestational diabetes and recommended to women with ovarian hyperstimulation syndrome (PCOS) to increase the chances of pregnancy or avoid early miscarriages. However, the exact effects of metformin on the female reproductive tract in general, and on the ovary in particular, are still not completely understood. In this study, we analyzed the effect of metformin on fertility and ovarian physiology in healthy female mice. We found that this drug altered the estrous cycle, early follicular development, serum estradiol and progesterone levels, and ovarian steroidogenic enzyme expression. Moreover, ovarian angiogenesis was lower in metformin-treated animals compared with untreated ones, whereas natural or gonadotropin-induced fertilization rates remained unchanged. However, offspring of metformin-treated animals displayed decreased body weight at birth. In this work, we unraveled the main effects of metformin on the ovary, isolated from other conditions such as hyperglycemia and hyperandrogenism, which is essential for a better understanding of metformin's mechanisms of action on reproduction and fertility.

High-Dose Neutrophil-Depleted Platelet-Rich Plasma Therapy for Knee Osteoarthritis: A Retrospective Study.

Background/Objectives: Encouraging results have been reported for Platelet-Rich Plasma (PRP) treatment for knee osteoarthritis (KOA). This study reports the efficacy and safety of a high dose of neutrophile and red-blood-cell-depleted PRP to treat patients with KOA. Methods: A total of 212 consecutive patients diagnosed with Kellgren-Lawrence (KL) grading 1-3 KOA chronic knee pain for at least 1 year were treated with three injections at 15-day intervals with a high dose of neutrophil-depleted PRP (4 billion platelets). Clinical outcomes were retrospectively recorded as the percentage of responders at 3-, 6-, and 12-month follow-up, following the OMERACT-OARSI criteria. Pain, through the VAS score and WOMAC score, was also been recorded. Results: A total of 4 mL of PRP containing 4 × 109 platelets was obtained by single-spin centrifugation and injected intra-articularly into each patient with no preactivation. The overall responder rate of patients responding to the OMERACT-OARSI criteria at 3, 6, and 12 months was 68.9%, 72.7%, and 70.6%, respectively. A significant improvement in VAS and WOMAC scores at 3-, 6-, and 12-month follow-up compared to the pretreatment value (p < 0.01) was observed. The lowest VAS score was observed at 6 months overall and in all three KL-graded groups. The KL2 groups showed the best results regarding pain reduction and their WOMAC score at 6 months (p < 0.01). Conclusions: For KL1-3 KOA, a high dosage of neutrophil-depleted PRP is a successful treatment. It has long-lasting effects that last up to one year, relieves symptoms, and may slow the advancement of the disease.

Cord blood transfusions in extremely low gestational age neonates to reduce severe retinopathy of prematurity: results of a prespecified interim analysis of the randomized BORN trial.

BACKGROUND: Preterm infants are at high risk for retinopathy of prematurity (ROP), with potential life-long visual impairment. Low fetal hemoglobin (HbF) levels predict ROP. It is unknown if preventing the HbF decrease also reduces ROP. METHODS: BORN is an ongoing multicenter double-blinded randomized controlled trial investigating whether transfusing HbF-enriched cord blood-red blood cells (CB-RBCs) instead of adult donor-RBC units (A-RBCs) reduces the incidence of severe ROP (NCT05100212). Neonates born between 24 and 27 + 6 weeks of gestation are enrolled and randomized 1:1 to receive adult donor-RBCs (A-RBCs, arm A) or allogeneic CB-RBCs (arm B) from birth to the postmenstrual age (PMA) of 31 + 6 weeks. Primary outcome is the rate of severe ROP at 40 weeks of PMA or discharge, with a sample size of 146 patients. A prespecified interim analysis was scheduled after the first 58 patients were enrolled, with the main purpose to evaluate the safety of CB-RBC transfusions. RESULTS: Results in the intention-to-treat and per-protocol analysis are reported. Twenty-eight patients were in arm A and 30 in arm B. Overall, 104 A-RBC units and 49 CB-RBC units were transfused, with a high rate of protocol deviations. A total of 336 adverse events were recorded, with similar incidence and severity in the two arms. By per-protocol analysis, patients receiving A-RBCs or both RBC types experienced more adverse events than non-transfused patients or those transfused exclusively with CB-RBCs, and suffered from more severe forms of bradycardia, pulmonary hypertension, and hemodynamically significant patent ductus arteriosus. Serum potassium, lactate, and pH were similar after CB-RBCs or A-RBCs. Fourteen patients died and 44 were evaluated for ROP. Ten of them developed severe ROP, with no differences between arms. At per-protocol analysis each A-RBC transfusion carried a relative risk for severe ROP of 1.66 (95% CI 1.06-2.20) in comparison with CB-RBCs. The area under the curve of HbF suggested that HbF decrement before 30 weeks PMA is critical for severe ROP development. Subsequent CB-RBC transfusions do not lessen the ROP risk. CONCLUSIONS: The interim analysis shows that CB-RBC transfusion strategy in preterm neonates is safe and, if early adopted, might protect them from severe ROP. TRIAL REGISTRATION: Prospectively registered at ClinicalTrials.gov on October 29, 2021. Identifier number NCT05100212.

Urban diabetes: analysis of diabetes prevalence in cities of the Lombardy region participating in the cities changing diabetes project.

AIM: The urban population increases by about 60 million people/year. Urbanization, unhealthy lifestyle and aging of the population are reflected in a constant growth in the prevalence of diabetes. In 2014, Steno Diabetes Centre in Copenhagen, University College London and Novo Nordisk, launched the Cities Changing Diabetes® program with the aim of creating a unified movement that would stimulate policy-makers to prioritize urban diabetes. METHODS: The socio-demographic data derive from (1) ISTAT (National Institute of Statistics of Italy), (2) ATS Metropolitan City of Milan, (3) ATS Val Padana-Cremona, (4) ATS Insubria-Varese, (5) The unemployment rates of the various municipalities have been extrapolated from an ISTAT-MEF elaboration published by Sole 24 Ore journal. RESULTS: In the different sanitary districts of the Metropolitan City of Milan, a strong linear correlation was found between the prevalence of diabetes and the prevalence of heart disease (R = 0.695, p < 0.001), as well as between the prevalence of diabetes and of nephropathies (R = 0.316, p < 0.001). The analysis concerning the province of Cremona showed a fair correlation between the prevalence of diabetes and cardiovascular disease (R = 0.658, p < 0.001). Even for the municipalities of Varese, the analysis documented a good correlation between the prevalence of diabetes and heart disease (R = 0.419, p < 0.001), but not between diabetes and nephropathies. CONCLUSIONS: Interesting differences in the relationship of diabetes prevalence with several diseases and socio-demographic factors have been found when comparing the metropolitan City of Milan with two smaller size cities as Varese and Cremona. Our present data confirm the hypothesis that urban diabetes will be the challenge for our society during the next decades.

Pure red cell aplasia among ABO mismatched hematopoietic stem cell transplant recipients: a 13-years retrospective study and literature review.

Background: Pure red cell aplasia (PRCA) is a possible complication after allogeneic hematopoietic stem cell transplantation (HSCT) with major ABO incompatibility. Patients experience delayed engraftment of the erythroid series, with prolonged transfusion-dependent anemia and iron overload. Methods: We performed a revision of the most recent literature about post-HSCT PRCA treatment procedures. Moreover, we conducted a retrospective study, over the last 13-years, which included all consecutive major ABO mismatched HSCT performed in our unit, with the aim to assess PRCA incidence, risk factors, and response to different treatments. Overall, 194 patients received a major ABO mismatched transplant from 2010 to 2022. For each patient, data about demographic and transplant characteristics, engraftment, blood transfusion, and possible treatment received were collected. Results: The literature review returned 23 eligible papers on PRCA treatment, with high success rate using plasma-exchange (PEX) and immunoadsorption procedures, daratumumab, and eltrombopag. Our study identified a total of 24 cases of PRCA. Among risk factors for PRCA development, we have found older recipient age (p=0.01), high pre-HSCT IgG and IgM IHA titer (p<0.0001), major rather than bidirectional ABO incompatibility (p=0.02), low T CD8 lymphocyte count in the graft (p=0.006), relative donor (p=0.02) and bone marrow as stem cell source (p=0.002). However, multivariate analysis confirmed only pre-HSCT IgG IHA titer as the unique risk factor for PRCA occurrence. The optimal cut-off value of pre-HSCT IgG IHA for PRCA development, resulted to be 1/64, with a 100% sensitivity and 68.8% specificity (p<0.0001). All patients with PRCA had received rhEPO and transfusion support and 20 patients received additional treatments like PEX, rituximab, and more recently daratumumab. Comprehensively, PEX and rituximab obtained a response in half of the cases, at a variable time, while the few cases of patients we treated with daratumumab suggest promising results. The overall response rate in our cohort was 75%, with significantly better survival (94.4% vs. 16.7%) and lower transplant-related mortality (6.3% vs. 80%) for PRCA responders. Conclusions: Standardized guidelines on when and how to treat PRCA are necessary because the current treatment is controversial among centers.

Experiencia de un programa de estandarización de creatinina en una región de ingresos bajos de Latinoamérica / Experience of a creatinine standardisation programme in a low-income region of Latin America / Experiência de um programa de padronização de creatinina em uma região de baixa renda da América Latina

Resumen Los programas de estandarización de creatinina mantienen su vigencia. El objetivo es describir la experiencia adquirida durante el desarrollo de un programa de estandarización de creatinina en una provincia de bajos recursos y mostrar los aspectos que se deben considerar para su escalabilidad en un contexto semejante. El programa se desarrolló en etapas: en la primera (2010) se realizó el relevamiento de 49 laboratorios clínicos (LC) distribuidos en toda la provincia del Chaco, Argentina. En la segunda (2012) se ajustó el error aleatorio (EA) aplicando protocolos internacionales (CLSI EP-5A). En la tercera etapa (2014-2015) se procesaron paneles de sueros con concentraciones asignadas por un método trazable al de referencia y al estándar internacional (CG-IDMS). Se aplicaron protocolos internacionales para evaluar el error total (ET) de la determinación en cada laboratorio (CLSI EP-10A). En 2016, aplicando herramientas de calidad, se evaluaron las barreras en el proceso. Se observó en el EA: para un nivel de 1,00 mg/dL, ningún LC alcanzó los niveles deseables; para un nivel de 2,5 mg/dL solo 9 (23%) los alcanzaron. Concluida la segunda y tercera etapa, solo 18 laboratorios (48,7%) lograron ajustar el EA y/o ET, pero resultó dificultoso sostenerlo en el tiempo. Los reactivos, calibradores y controles son producidos por la industria y depende del estado el control de los mismos. La homogeneidad del equipamiento depende de la accesibilidad económica y del volumen de trabajo. El medio ambiente, la temperatura y la calidad del agua siguen siendo una dificultad para la escalabilidad.

Abstract Creatinine standardisation programmes remain valid. The objective of this work is to describe the experience acquired during the development of a creatinine standardisation programme in a low-resource province and show the aspects that should be considered for its scalability in a similar context. The programme was developed in stages. The first one was carried out in 2010. It consists of a structured survey completed by 49 clinical laboratories (CL) distributed throughout the province. In the second stage (2012) the random error (RE) was adjusted by applying international protocols (CLSI EP-5A). In the third stage (2014-2015), panels of sera were processed with concentrations assigned by a method traceable to the reference and the international standard (CG-IDMS). International protocols were applied to evaluate the total error (TE) of the determination in each laboratory (CLSI EP-10A). In 2016, applying quality tools, the barriers in the process were evaluated. In the RE, it was observed: for a level of 1.00 mg/dL, no CL reached the desirable levels; for a level of 2.5 mg/dL only 9 (23%) CL achieved them. Once the second and third stages were completed, only 18 laboratories (48.7%) managed to adjust the RE and/or TE, but it was difficult to sustain it over time. With respect to materials, reagents, calibrators, and controls, they are produced by the industry depending on the state of their control. The homogeneity of the equipment depends on economic accessibility and volume of work. The environment, temperature, and water quality are a barrier to scalability.

Resumo Os programas de padronização da creatinina permanecem válidos. O objetivo é descrever a experiência adquirida durante o desenvolvimento de um programa de padronização de creatinina em uma província com poucos recursos e mostrar os aspectos que devem ser levados em consideração para sua escalabilidade em um contexto semelhante. O programa foi desenvolvido em etapas: Na primeira (2010), foi realizado um levantamento de 49 laboratórios clínicos (LC) distribuídos em toda a provincia do Chaco, na Argentina. Na segunda etapa (2012) o erro aleatório (EA) foi ajustado através da aplicação de protocolos internacionais (CLSI EP-5A). Na terceira etapa (2014-2015), foram processados paineis de soros com concentrações atribuídas por método rastreável à referência e ao padrão internacional (CG-IDMS). Protocolos internacionais foram aplicados para avaliar o erro total (ET) da determinação em cada laboratório (CLSI EP-10A). Em 2016, aplicando ferramentas de qualidade, foram avaliadas as barreiras no processo. Observou-se na EA: para o nível de 1,00 mg/dL nenhuma LC atingiu os níveis desejáveis; para um nível de 2,5 mg/dL, apenas 9 (23%) os atingiram. Concluídas a segunda e terceira etapas, apenas 18 laboratórios (48,7%) conseguiram ajustar o EA e/ou o ET, mas foi difícil sustentá-lo ao longo do tempo. No que diz respeito aos reagentes, calibradores e controles, eles são produzidos pela indústria. dependendo do estado o seu controle. A homogeneidade do equipamento depende da acessibilidade econômica e do volume de trabalho. O meio ambiente, a temperatura e a qualidade da água continuam sendo uma dificuldade para a escalabilidade.

Unselected donor-derived hematopoietic stem cells boost for Chimeric Antigen Receptor T-cell associated hematotoxicity.

Hematological toxicity following Chimeric Antigen Receptor-T therapy in a patient with a prior allogeneic stem cell transplantation was resolved by the infusion of unselected donor-derived stem cell boost. Due to the donor's lymphocytes, the patient experienced a well-controlled flare-up of acute graft versus host disease.

Hyposalivation and periodontal disease as oral non-articular characteristics in rheumatoid arthritis.

OBJECTIVE: To investigate the association among rheumatoid arthritis (RA), saliva production, and periodontal status. METHODS: An observational study was carried out on 103 subjects with RA and 103 without RA matched by sex and age. Rheumatologic evaluation included serological and clinical variables. A full mouth periodontal examination was performed according to the American Academy of Periodontology (1999). Resting and stimulated whole salivary flows were determined after spiting during 5 min. RESULTS: RA was associated with a higher prevalence of severe periodontitis (12% vs. 4%), with a marked reduction in resting and stimulated saliva production, and with a higher prevalence of resting (19% vs. 0%) and also stimulated hyposalivation (54% vs. 10%), compared with the control group. The differences in mean resting and stimulated salivary flows between RA and control groups persisted after the exclusion of people with hyposalivation. Saliva production was not associated with the presence or the severity of periodontal disease, or with the rheumatic clinical characteristics of the patients. CONCLUSIONS: More than 50% of people with RA have some degree of reduction in their salivary flows, an affection not associated with the periodontal status or rheumatic activity, which are the expression of the two related inflammatory diseases. The influence of autonomic dysfunction on hyposalivation can be considered. While periodontitis would be a disease-associated comorbidity of RA, poor saliva production should be included among the extra-articular manifestations. Key Points • Rheumatoid arthritis patients are more prone to suffer from periodontitis and/or hyposalivation. • Periodontal disease is more prevalent in people with rheumatoid arthritis and also an association was found between the severities of both pathologies. • More than 50% of people with RA would have some degree of reduction in their salivary flows, an affection not associated with the periodontal status or rheumatic activity. • Reduced saliva production in rheumatoid arthritis patients should be included among the extra-articular manifestations.

The application of a multidisciplinary approach in the diagnosis of Castleman disease and Castleman-like lymphadenopathies: A 20-year retrospective analysis of clinical and pathological features.

BACKGROUND: Castleman disease (CD) comprises a group of rare and heterogeneous haematological disorders, including unicentric (UCD) and multicentric (MCD) forms, the latter further subdivided into HHV8-MCD, POEMS-MCD and idiopathic-MCD (iMCD). However, according to the Castleman Disease Collaborative Network guidelines, the diagnosis of CD can only be achieved through collaboration between clinicians and pathologists. METHODS: We applied these clinical and pathological criteria and implement with clonality testing to a retrospective cohort of 48 adult and paediatric Italian patients diagnosed with reactive lymphadenitis with CD-like histological features. RESULTS: We confirmed the diagnosis of CD in 60% (29/48) of the cases, including 12 (41%) UCD and 17 (59%; five HHV8-MCD, three POEMS-MCD and nine iMCD) MCD. Of the remaining 19 cases (40%) with multiple lymphadenopathy, 5 (26%) were classified as autoimmune diseases, 1 (5%) as autoimmune lymphoproliferative disorder, 1 (5%) as IgG4-related disease, 11 (83%) as reactive lymphadenitis and 1 (5%) as nodal marginal zone lymphoma. CONCLUSIONS: Our study emphasizes the importance of the multidisciplinary approach to reactive lymphadenitis with CD-like features in order to achieve a definitive diagnosis and choose the appropriate treatment.

Correction: Bocci et al. Remarkable Remission Rate and Long-Term Efficacy of Upfront Metronomic Chemotherapy in Elderly and Frail Patients, with Diffuse Large B-Cell Lymphoma. J. Clin. Med. 2022, 11, 7162.

In the original publication [...].

Curcumin nanocrystals-in-nanofibres as a promising platform for the management of periodontal disease.

It is estimated that nearly a half of the world's population over 30 years old suffer from some kind of periodontal disease (PD). Although preventable, PD can pose a significant health burden to patients, causing from pain and discomfort to disfigurement and death. The management of PD often requires surgical procedures accompanied of systemic antibiotic and anti-inflammatory treatments. Curcumin (CUR), a potent anti-inflammatory and antimicrobial active, has shown great promise in the management of PD; however, its effects are often limited by its low bioavailability. In this work, we report the development of electrospun nanofibres (NFs) loaded with CUR nanocrystals (NCs) for the management of PD. NCs of 100 nm were obtained by media milling and loaded into dissolving polyvinyl alcohol NFs using electrospinning. The resultant NCs-in-NFs dissolved in water spontaneously, releasing NCs with a particle size of âˆ¼120 nm. The physiochemical characterisation of the systems indicated the absence of chemical interactions between drug and polymer, and nanofibres with an amorphous nature. In vitro release profiles demonstrated that the NCs had a significantly higher dissolution rate (∼100 % at day 40) than the control group (approximately 6 % at day 40), which consisted of NFs containing a physical mixture of the drug and stabiliser. Finally, mucosal deposition studies demonstrated a 10-fold higher capacity of the novel NCs-in-NFs system to deposit CUR ex vivo using excised neonatal porcine mucosal tissue, when compared to the control group.

Allogenic Cord Blood Transfusion in Preterm Infants.

Repeated red blood cell (RBC) transfusions in preterm neonates cause the progressive displacement of fetal hemoglobin (HbF) by adult hemoglobin. The ensuing increase of oxygen delivery may result at the cellular level in a dangerous condition of hyperoxia, explaining the association between low-HbF levels and retinopathy of prematurity or bronchopulmonary dysplasia. Transfusing preterm neonates with RBC concentrates obtained from allogeneic umbilical blood is a strategy to increase hemoglobin concentration without depleting the physiologic HbF reservoir. This review summarizes the mechanisms underlying a plausible beneficial impact of this strategy and reports clinical experience gathered so far in this field.

IMT504 protects beta cells against apoptosis and maintains beta cell identity, without modifying proliferation.

We have demonstrated that oligodeoxynucleotide IMT504 promotes significant improvement in the diabetic condition in diverse animal models. Based on these results, here we evaluated whether these effects observed in vivo could be due to direct effects on ß-cells. We demonstrate by immunofluorescence that IMT504 enters the cell and locates in cytoplasm where it induces GSK-3ß phosphorylation that inactivates this kinase. As GSK-3ß tags Pdx1 for proteasomal degradation, by inactivating GSK-3ß, IMT504 induces an increase in Pdx1 protein levels, demonstrated by Western blotting. Concomitantly, an increase in Ins2 and Pdx1 gene transcription was observed, with no significant increase in insulin content or secretion. Enhanced Pdx1 is promising since it is a key transcription factor for insulin synthesis and is also described as an essential factor for the maintenance ß-cell phenotype and function. Dose-dependent inhibition of H2 O2 -induced apoptosis determined by ELISA as well as decreased expression of Bax was also observed. These results were confirmed in another ß-cell line, beta-TC-6 cells, in which a cytokine mix induced apoptosis that was reversed by IMT504. In addition, an inhibitor of IMT504 entrance into cells abrogated the effect IMT504. Based on these results we conclude that the ß-cell recovery observed in vivo may include direct effects of IMT504 on ß-cells, by maintaining their identity/phenotype and protecting them from oxidative stress and cytokine-induced apoptosis. Thus, this work positions IMT504 as a promising option in the framework of the search of new therapies for type I diabetes treatment.

Hypnosis to reduce fear of falling in hospitalized older adults: a feasibility randomized controlled trial.

BACKGROUND: Fear of falling is associated with numerous negative health outcomes in older adults and can limit the rehabilitation process. Hypnosis is now recognized as an effective treatment for a variety of conditions, especially anxiety and pain, which can be integrated safely with conventional medicine. The objective of this study was to assess the feasibility and acceptability of a hypnosis intervention in hospitalized older adults to reduce fear of falling. METHODS: In this feasibility randomized controlled trial, 32 older patients, hospitalized in geriatric rehabilitation wards, were randomly allocated (1:1 ratio) to either an intervention group (hypnosis, 2 sessions, one per week, plus usual rehabilitation program) or a control group (usual rehabilitation program only). Clinical assessors and statistician were blinded to group allocation. Primary outcomes were recruitment rate, retention rate, and adherence to the intervention. Exploratory outcomes, analyzed according to the intention-to-treat principle, included impact of hypnosis on fear of falling (assessed by a new scale perform-FES), functional status, in-hospital falls, and length of hospital stay. RESULTS: Recruitment rate was 1.3 patients per week. The recruitment of the population sample was achieved in 5.5 months. The retention rate did not differ significantly between groups and a good adherence to the hypnosis intervention was achieved (77% of patients received the full intervention). No adverse event related to the hypnosis intervention was observed. Regarding exploratory clinical outcomes, no differences were found between groups on any outcome. CONCLUSION: Hypnosis is feasible and well accepted in a geriatric hospitalized population undergoing rehabilitation. Further pilot work should be conducted, with an increased number of hypnosis sessions, before conducting a full-scale trial to conclude whether, or not, hypnosis is effective to reduce fear of falling. TRIAL REGISTRATION: NCT04726774.