Characterization of a Novel Approach for Neonatal Hematocrit Screening Based on Penetration Velocity in Lateral Flow Test Strip.

Hematocrit (HCT) is a crucial parameter for both adult and pediatric patients, indicating potentially severe pathological conditions. Most common methods for HCT assessment are microhematocrit and automated analyzers; however, developing countries present specific needs often not addressed by these technologies. Paper-based devices can be suitable for those environments being inexpensive, rapid, easy to use, and portable. The aim of this study is to describe and validate against a reference method, a novel HCT estimation method based on penetration velocity in lateral flow test strips complying with the requirements in low- or middle-income country (LMIC) scenarios. To calibrate and test the proposed method, 145 blood samples of 105 healthy neonates with gestational age greater than 37 weeks were collected (29 calibration set, 116 test set) in the range of HCT values (31.6-72.5%). The time difference (Δt) from the whole blood sample loading into the test strip instant till the nitrocellulose membrane saturation instant was measured by a reflectance meter. A nonlinear relation was observed between HCT and Δt and was estimated by a third-degree polynomial equation (R2 = 0.91) valid in 30% to 70% HCT interval. The proposed model was subsequently used to estimate HCT values on the test set showing a good agreement between the estimated HCT and the HCT measured by the reference method (r = 0.87, p < 0.001), with a low mean difference of 0.53 ± 5.04% and a slight trend of overestimation for higher hematocrit values. The mean absolute error was 4.29%, while the maximum absolute error was 10.69%. Although the proposed method did not present a sufficient accuracy to be used for diagnostic purposes, it could be suitable as a fast, low-cost, easy-to-use screening tool especially in LMIC scenarios.

Applications of new radiological scores: the Node-rads in colon cancer staging.

PURPOSE: The study focuses on the evaluation of the new Node Reporting and Data System 1.0 (Node-rads) scoring accuracy in the assessment of metastatic lymph nodes (LN) in patients with colon carcinoma. MATERIAL AND METHODS: From April 2021 to May 2022, retrospective chart reviews were performed on 67 preoperative CT (Computed Tomography) of patients undergoing excisional surgery for colon cancer at the Polyclinic of Bari, Italy. Primary endpoints were to assess lymph node size and configuration to express the likelihood of a metastatic site adopting the Node-rads score system, whose categories of risk are defined from 1 (very low) to 5 (very high). The nodal postsurgical histological evaluation was the gold standard. The relationship between Node-rads score, LN size, configuration criteria (texture, border and shape) and the presence of histological metastases was statistically evaluated. RESULTS: All surgical specimens examined had correlation with Node-rads score. They were significantly more likely to present nodes micrometastasis those patients with (a) spherical LN shape (82.8%), (b) with lymph node necrosis (100%), (c) irregular borders (87%) and (d) the LN short axis more than 10 mm (61.9%). CONCLUSIONS: Our experience highlights how the Node-rads system proposes an intuitive and effective definition of criteria to standardize the lymph node radiological reports in colon cancer disease. Further studies are needed to streamline the classification of the nodal and peripheral LN in all the oncological imaging.

Synthesis and Antibacterial Evaluation of New Pyrazolo[3,4-d]pyrimidines Kinase Inhibitors.

Pyrazolo[3,4-d]pyrimidines represent an important class of heterocyclic compounds well-known for their anticancer activity exerted by the inhibition of eukaryotic protein kinases. Recently, pyrazolo[3,4-d]pyrimidines have become increasingly attractive for their potential antimicrobial properties. Here, we explored the activity of a library of in-house pyrazolo[3,4-d]pyrimidines, targeting human protein kinases, against Staphylococcus aureus and Escherichia coli and their interaction with ampicillin and kanamycin, representing important classes of clinically used antibiotics. Our results represent a first step towards the potential application of dual active pyrazolo[3,4-d]pyrimidine kinase inhibitors in the prevention and treatment of bacterial infections in cancer patients.

Clinical aspects of Fanconi anemia individuals with the same mutation of FANCF identified by next generation sequencing.

BACKGROUND: Fanconi anemia (FA) is a rare genetic disease characterized by congenital malformations, aplastic anemia and increased risk of developing malignancies. FA is genetically heterogeneous as it is caused by at least 17 different genes. Among these, FANCA, FANCC, and FANCG account for approximately 85% of the patients whereas the remaining genes are mutated in only a small percentage of cases. For this reason, the molecular diagnostic process is complex and not always extended to all the FA genes, preventing the characterization of individuals belonging to rare groups. METHODS: The FA genes were analyzed using a next generation sequencing approach in two unrelated families. RESULTS: The analysis identified the same, c.484_485del, homozygous mutation of FANCF in both families. A careful examination of three electively aborted fetuses in one family and one affected girl in the other indicated an association of the FANCF loss-of-function mutation with a severe phenotype characterized by multiple malformations. CONCLUSION: The systematic use of next generation sequencing will allow the recognition of individuals from rare complementation groups, a better definition of their clinical phenotypes, and consequently, an appropriate genetic counseling.

Molecular analysis of Fanconi anemia: the experience of the Bone Marrow Failure Study Group of the Italian Association of Pediatric Onco-Hematology.

Fanconi anemia is an inherited disease characterized by congenital malformations, pancytopenia, cancer predisposition, and sensitivity to cross-linking agents. The molecular diagnosis of Fanconi anemia is relatively complex for several aspects including genetic heterogeneity with mutations in at least 16 different genes. In this paper, we report the mutations identified in 100 unrelated probands enrolled into the National Network of the Italian Association of Pediatric Hematoly and Oncology. In approximately half of these cases, mutational screening was carried out after retroviral complementation analyses or protein analysis. In the other half, the analysis was performed on the most frequently mutated genes or using a next generation sequencing approach. We identified 108 distinct variants of the FANCA, FANCG, FANCC, FANCD2, and FANCB genes in 85, 9, 3, 2, and 1 families, respectively. Despite the relatively high number of private mutations, 45 of which are novel Fanconi anemia alleles, 26% of the FANCA alleles are due to 5 distinct mutations. Most of the mutations are large genomic deletions and nonsense or frameshift mutations, although we identified a series of missense mutations, whose pathogenetic role was not always certain. The molecular diagnosis of Fanconi anemia is still a tiered procedure that requires identifying candidate genes to avoid useless sequencing. Introduction of next generation sequencing strategies will greatly improve the diagnostic process, allowing a rapid analysis of all the genes.

Focal dermal hypoplasia (Goltz-Gorlin syndrome): a new case with a novel variant in the PORCN gene (c.1250T>C:p.F417S) and unusual spinal anomaly.

Focal dermal hypoplasia (FDH; Goltz-Gorlin syndrome; OMIM 305600) is a disorder that features involvement of the skin, skeletal system, and eyes. It is caused by loss-of-function mutations in the PORCN gene. We report a young girl with FDH, microphthalmos associated with colobomatous orbital cyst, dural ectasia and cystic malformation of the spinal cord, and a de novo variant in PORCN. This association has not been previously reported, and based on these observations the phenotypic spectrum of FDH might be broader than previously appreciated. It would be prudent to alter the suggested surveillance for this rare disorder.

Do social media interventions increase vaccine uptake?

Introduction: The Italian mass COVID-19 vaccination campaign has included children aged 5-11 years as part of the target population since December 2021. One of the biggest challenges to vaccine uptake was vaccine hesitancy among parents and children's caregivers. Primary care pediatricians (PCPs), as the first point of contact between the National Health Service (NHS) and parents/caretakers, initiated various communication strategies to tackle this hesitancy. This study aims to evaluate the impact of a PCP-led social media intervention and a digital reminder service (DRS) on parental hesitancy regarding vaccinating their 5-11-year-old children against COVID-19. Methods: A prospective cohort study was designed, and the chosen target populations were parents and caretakers of children aged 5-11 years. Two PCP cohorts were recruited. The first group received a social media intervention and a DRS; while the second group did not. Both cohorts had access to traditional face-to-face and telephone-based counseling. The vaccination coverage rate in the two groups was evaluated. Results: A total of 600 children were enrolled. The exposed cohort (277 patients) received social media intervention, DRS, and counseling options (face-to-face and telephone-based), whereas the non-exposed cohort (323 patients) received only counseling options. In total, 89 patients from the exposed cohort did not receive any dose of the COVID-19 vaccine (32.5%), 165 were fully immunized (59.5%), and 23 received only one dose (8.5%). A total of 150 non-exposed patients did not receive any dose of the COVID-19 vaccine (47%), 147 were fully immunized (45.5%), and 24 only received one dose (7.4%). The difference between the two groups was statistically significant (chi square = 11.5016; p = 0.0006). Conclusion: Social media and DRS interventions had a positive impact on vaccine uptake and may be helpful in tackling vaccine hesitancy. Better-designed studies are needed to corroborate these findings.

Social Media Interventions Strengthened COVID-19 Immunization Campaign.

Background: Since The Italian Medicines Agency (AIFA) has recommended the COVID-19 vaccine Comirnaty in children aged 5-11, the immunization campaign faced vaccine hesitancy in parents. Social media are emerging as leading information source that could play a significant role to counteract vaccine hesitancy, influencing parents' opinions and perceptions. Our aim was to evaluate the coverage of the COVID-19 vaccine Comirnaty in a cohort of children aged 5-11 whose families have been counseled to use Social Media to counteract vaccine hesitancy. Methods: All parents of children aged 5-11 in a primary care setting were instructed by their pediatrician to get accurate information about the COVID-19 vaccine from a Facebook page. Active calls to vaccinate children were also scheduled through messaging services Pediatotem and Whatsapp. Vaccination rates of children in the study were assessed with an electronic database and compared to both regional and national child vaccination rates. Results: Coverage of 277 children aged 5-11 was analyzed from 16 December 2021 to 31 January 2022. A total of 62.4% (173/277) of enrolled children received the 1st dose of COVID-19 vaccine Comirnaty and 39.7% (110/277) the 2nd dose. Coverage rates were higher compared both to the regional population (1st dose: 48.8%, 2nd dose: 24.6%; p = 0.001) and national population (1st dose: 32.1%, 2nd dose: 13.8%; p < 0.001). Conclusion: Increasing vaccine confidence using Social Media interventions have a positive impact on vaccination acceptance of parents.

COVID-19 in Italy: Comparison of CT Findings from Time Zero to the Delta Variant.

On 12 March 2020, the World Health Organization (WHO) declared the novel Coronavirus (CoV) disease a global Pandemic and an emerging risk. In order to understand patterns that are typical in COVID-19 pneumonia and track the evolution of the disease, the role of the chest computed tomography (CT) is pivotal. The impact of the illness as well as the efficiency of the therapy are also monitored carefully when performing this imaging exam. Coronaviruses, specifically CoV-2, as RNA viruses, have a tendency to frequently change their genome, giving the virus beneficial characteristics such as greater transmissibility, pathogenicity and the possibility to escape the previously acquired immunity. Therefore, genome evaluation became an extremely important routine practice worldwide. In particular, in Italy, four variants have been recognised and each of them represent a specific temporal wave of the disease. Hence, our goal was to describe imaging findings of COVID-19 pneumonia, specifically its most typical imaging identified during the period of our study, and to assess whether or not SARS-CoV-2 variants determine different CT patterns. Our analyses revealed that the SARS-CoV-2 genotype seems not to interfere with the severity of CT patterns and, in particular, bilateral Ground Glass Opacities (GGOs) are the most frequent findings in all COVID-19 waves.

Impact of six years of routine varicella vaccination on the disease-related hospitalizations at Minas Gerais, Brazil.

INTRODUCTION: The varicella vaccine was first introduced into the Brazilian immunization schedule in September 2013 as a single dose for children aged 15 months. In 2018, a second dose was recommended for individuals between 4 and 6 years old. This study aims to assess the impact of routine varicella vaccination on the number and profile of hospitalized varicella patients during the single dose period, as well as in the first two years after the adoption of the second dose. METHODS: An observational retrospective study was conducted in an infectious disease pediatric hospital, in Minas Gerais, Brazil. Clinical as well as epidemiological data from patients hospitalized due to varicella between 2010 and 2019 were collected. Patients were split into groups based on the vaccine introduction: pre-vaccine period, single dose and two-dose period. They were compared by age, sex, reason for admission, illness-related complications and clinical outcome. RESULTS: There were 1193 admissions due to varicella during the studied period. When compared with the pre-vaccine period, the number of hospitalizations decreased in 61.5% during the single-dose regime, reaching 95.2% in the two-dose period. Hospitalization rates decreased in all age groups, including non-vaccinated individuals such as those younger than 12 months (92.1%). As for reasons of admission, secondary bacterial skin infections were perceived to be the most common cause (>70%). A reduction was also seen in admission of immunocompromised or HIV positive patients (84.8%). CONCLUSION: The collected data shows a significant impact in the number of hospital admissions due to varicella after six years of the implementation of the vaccine, positively affecting both vaccinated and non-vaccinated individuals. Further reduction was seen after the second dose was initiated, but its true impact will only be understood fully after a longer period of continuous vaccination.

Insights into RNA-dependent RNA Polymerase Inhibitors as Antiinfluenza Virus Agents.

BACKGROUND: Influenza is a seasonal disease that affects millions of people every year and has a significant economic impact. Vaccines are the best strategy to fight this viral pathology, but they are not always available or administrable, prompting the search for antiviral drugs. RNA-dependent RNA polymerase (RdRp) recently emerged as a promising target because of its key role in viral replication and its high conservation among viral strains. DISCUSSION: This review presents an overview of the most interesting RdRp inhibitors that have been discussed in the literature since 2000. Compounds already approved or in clinical trials and a selection of inhibitors endowed with different scaffolds are described, along with the main features responsible for their activity. RESULTS: RdRp inhibitors are emerging as a new strategy to fight viral infections and the importance of this class of drugs has been confirmed by the FDA approval of baloxavir marboxil in 2018. Despite the complexity of the RdRp machine makes the identification of new compounds a challenging research topic, it is likely that in the coming years, this field will attract the interest of a number of academic and industrial scientists because of the potential strength of this therapeutic approach.

In-hospital and web-based intervention to counteract vaccine hesitancy in very preterm infants' families: a NICU experience.

BACKGROUND: Vaccine hesitancy is a global problem, carrying significant health risks for extremely vulnerable population as that of preterm infants. Social media are emerging as significant tools for public health promotion. Our aim was to evaluate both the coverage and the timeliness of routine immunizations in a cohort of preterm infants (< 33 weeks of gestational age) at 24 months of age whose families have been subjected to in-hospital and web-based interventions to counteract vaccine hesitancy. METHODS: For a period of 2 years parents of preterm infants were instructed during their follow up visits by a member of the NICU team to get correct informations about vaccines from a social network page. Vaccination rates of preterm infants were assessed at 24 months of chronological age with an electronic database and compared to both general population and historical cohort. RESULTS: Coverage and timeliness of vaccinations at 24 months of age of 170 preterm infants were analyzed in December 2019. Gestational age and birth weight median (IQR) were, respectively, 31.0 (5.0) weeks and 1475.0 (843.8) g. Coverage rates were similar to those of the regional population (p > 0.05), while timeliness of administration was significantly delayed compared to the recommended schedule (p < 0.001). Age of administration was not correlated with either body weight and gestational age at birth (Spearman rank, p > 0.05). DTaP-IPV-HBV-Hib 2nd and 3rd doses, MMR and Varicella vaccines coverage data were higher compared to historical cohort (p < 0.05). CONCLUSION: Increasing vaccine confidence through web-based interventions could have a positive impact on vaccination acceptance of parents of preterm infants, although timeliness results still delayed. There is a strong need to develop different and effective vaccination strategies to protect this very vulnerable population.

Development of Pyrazolo[3,4-d]pyrimidine Kinase Inhibitors as Potential Clinical Candidates for Glioblastoma Multiforme.

Glioblastoma multiforme (GBM) is the most aggressive primary brain tumor. Residual cells at the tumor margin are responsible for up to 85% of GBM recurrences after standard treatment. Despite this evidence, the identification of compounds active on this cell population is still an underexplored field. Herein, starting from the knowledge that kinases are implicated in GBM, we evaluated three in-house pyrazolo[3,4-d]pyrimidines active as Src, Fyn, and SGK1 kinase inhibitors against patient derived cell lines from either the invasive region or contrast-enhanced core of GBM. We identified our Src inhibitor, SI306, as a promising lead compound for eradicating invasive GBM cells. Furthermore, aiming at the development of a feasible oral treatment for GBM, we performed a formulation study using 2D inkjet printing to generate soluble polymer-drug dispersions. Overall, this study led to the identification of a set of polymer-formulated pyrazolo[3,4-d]pyrimidine kinase inhibitors as promising candidates for GBM preclinical efficacy studies.

An Update on JAK Inhibitors.

Janus kinases (JAKs) are a family of non-receptor tyrosine kinases, composed by four members, JAK1, JAK2, JAK3 and TYK2. JAKs are involved in different inflammatory and autoimmune diseases, as well as in malignancies, through the activation of the JAK/STAT signalling pathway. Furthermore, the V617F mutation in JAK2 was identified in patients affected by myeloproliferative neoplasms. This knowledge prompted researchers from academia and pharmaceutical companies to investigate this field in order to discover small molecule JAK inhibitors. These efforts recently afforded to the market approval of four JAK inhibitors. Despite the fact that all these drugs are pyrrolo[2,3-d]pyrimidine derivatives, many compounds endowed with different heterocyclic scaffolds have been reported in the literature as selective or multi-JAK inhibitors, and a number of them is currently being evaluated in clinical trials. In this review we will report many representative compounds that have been published in articles or patents in the last five years (period 2013-2017). The inhibitors will be classified on the basis of their chemical structure, focusing, when possible, on their structure activity relationships, selectivity and biological activity. For every class of derivatives, compounds disclosed before 2013 that have entered clinical trials will also be briefly reported, to underline the importance of a particular chemical scaffold in the search for new inhibitors.

Identification of a new family of pyrazolo[3,4-d]pyrimidine derivatives as multitarget Fyn-Blk-Lyn inhibitors active on B- and T-lymphoma cell lines.

Abnormal activation of B-cell receptor (BCR) signaling plays a key role in the development of lymphoid malignancies, and could be reverted by the simultaneous inhibition of Lyn, Fyn and Blk, three members of the Src family kinase (SFK). Fyn and Blk are also promising targets for the treatment of some forms of T-cell non-Hodgkin lymphoma which point to the druggability of SFKs for the treatment of these cancers. We recently identified Si308 as a potent Fyn inhibitor, while preliminary data showed that it might also inhibit Lyn and Blk. Here, molecular modelling studies were coupled with enzymatic assays to further investigate the effect of Si308 on Lyn and Blk. A small library of pyrazolo[3,4-d]pyrimidines structurally related to Si308 was synthesized and tested on human lymphoma cell lines. Compound 2h emerged as a new multitarget inhibitor of Lyn, Fyn and Blk endowed with remarkable antiproliferative effects on human B and T lymphoma cell lines. Its favorable ADME properties make the compound suitable for further developments.

Recent Studies on Ponatinib in Cancers Other Than Chronic Myeloid Leukemia.

Ponatinib is a third line drug for the treatment of chronic myeloid leukemia patients, especially those that develop the gatekeeper mutation T315I, which is resistant to the first and the second line drugs imatinib, nilotinib, dasatinib and bosutinib. The compound was first identified as a pan Bcr-Abl and Src kinase inhibitor. Further studies have indicated that it is a multitargeted inhibitor that is active on FGFRs, RET, AKT, ERK1/2, KIT, MEKK2 and other kinases. For this reason, the compound has been evaluated on several cancers in which these kinases play important roles, including thyroid, breast, ovary and lung cancer, neuroblastoma, rhabdoid tumours and in myeloproliferative disorders. Ponatinib is also being tested in clinical trials to evaluate its activity in FLT3-ITD acute myelogenous leukemia, head and neck cancers, certain type of lung cancer, gastrointestinal stromal tumours and other malignancies. In this review we report the most recent preclinical and clinical studies on ponatinib in cancers other than CML, with the aim of giving a complete overview of this interesting compound.

Diagnostic Performance Analysis of the Point-of-Care Bilistick System in Identifying Severe Neonatal Hyperbilirubinemia by a Multi-Country Approach.

IMPORTANCE: The real prevalence and clinical burden of severe neonatal jaundice are undefined due to difficulties in measuring total serum bilirubin (TSB) outside secondary and tertiary clinical centers. OBJECTIVE: To assess the diagnostic performance of the point-of care Bilistick System (BS) in identifying neonatal jaundice patients requiring treatment. DESIGN: Between April 2015 and November 2016, 1911 neonates, were recruited to participate in the study. Blood samples were simultaneously collected for the TSB determination by BS and by hospital laboratory (Lab). Data were collected and sent to the Bilimetrix headquarter in Trieste where statistical analysis was performed. Newborns with neonatal jaundice were treated with phototherapy according to each center's guidelines. SETTING: 17 hospitals from Nigeria, Egypt, Indonesia, and Viet Nam. PARTICIPANTS: 1911 newborns were included, of which 1458 (76·3%) fulfilled the inclusion criteria. RESULTS: TSB level measured by BS agreed (p < .0001) with the lab result in all four countries. The diagnostic performance of BS showed a positive predictive value (PPV) of 92·5% and a negative predictive value (NPV) of 92·8%. CONCLUSIONS AND RELEVANCE: BS is a reliable system to detect neonatal jaundice over a wide range of bilirubin levels. Since Bilistick is a point-of-care test, its use may provide appropriate and timely identification of jaundiced newborns requiring treatment.

Pyrrolo[2,3-d]pyrimidines active as Btk inhibitors.

INTRODUCTION: Btk is a tyrosine kinase dysregulated in several B-cell malignancies and autoimmune diseases, and this has given rise to a search for Btk inhibitors. Nevertheless, only one Btk inhibitor, ibrutinib, has been approved to date, although other compounds are currently being evaluated in clinical trials or in preclinal stages. Area covered: This review, after a brief introduction on Btk and its inhibitors already in clinical trials, focusses on pyrrolo[2,3-d]pyrimidine derivatives patented in the last five years as Btk inhibitors. Indeed, the pyrrolo[2,3-d]pyrimidine scaffold, being a deaza-isostere of adenine, the nitrogenous base of ATP, is an actively pursued target for Btk inhibitors. The patent literature since 2012 have been extensively investigated, pointing out the general features of the patented compounds and, when it is possible, their mechanism of action. Expert opinion: The recently patented pyrrolo[2,3-d]pyrimidines, acting as reversible or irreversible inhibitors, showed a very interesting in vitro activity. For this reason, the development of compounds endowed with this scaffold could afford a significant impact in the search for drug candidates for the treatment of immune diseases or B-cell malignancies.

Asthma and Food Allergy in Children: Is There a Connection or Interaction?

This review explores the relationship between food allergy and asthma. They can share the same risk factors, such as parental allergy, atopic eczema, and allergen sensitization, and they often coincide in the same child. Coexistence may negatively influence the severity of both conditions. However, it remains to be determined whether food allergy may directly affect asthma control. An early food sensitization in the first year of life can predict the onset of asthma. Furthermore, asthmatic symptoms could rarely be caused by ingestion or inhalation of the offending food. Asthma caused by food allergy is severe and may be associated with anaphylactic symptoms. Therefore, an accurate identification of the offending foods is necessary in order to avoid exposure. Patients should be instructed to treat asthmatic symptoms quickly and to use self-injectable epinephrine.