The Microbial Diversity and Biofilm Characteristics of d-PTFE Membranes Used for Socket Preservation: A Randomized Controlled Clinical Trial

Background: Understanding microbial colonization on different membranes is critical for guided bone regeneration procedures such as socket preservation, as biofilm formation may affect healing and clinical outcomes. This randomized controlled clinical trial (RCT) investigates, for the first time, the microbiome of two different high-density polytetrafluoroethylene (d-PTFE) membranes that are used in socket preservation on a highly molecular level and in vivo. Methods: This RCT enrolled 39 participants, with a total of 48 extraction sites, requiring subsequent implant placement. Sites were assigned to two groups, each receiving socket grafting with a composite bone graft (50% autogenous bone, 50% bovine xenograft) and covered by either a permamem® (group P) or a Cytoplast™ (group C). The membranes were removed after four weeks and analyzed using scanning electron microscopy (SEM) for bacterial adherence, qPCR for bacterial species quantification, and next-generation sequencing (NGS) for microbial diversity and composition assessment. Results: The four-week healing period was uneventful in both groups. The SEM analysis revealed multispecies biofilms on both membranes, with membranes from group C showing a denser extracellular matrix compared with membranes from group P. The qPCR analysis indicated a higher overall bacterial load on group C membranes. The NGS demonstrated significantly higher alpha diversity on group C membranes, while beta diversity indicated comparable microbiota compositions between the groups. Conclusion: This study highlights the distinct microbial profiles of two d-PTFE membranes during the four-week socket preservation period. Therefore, the membrane type and design do, indeed, influence the biofilm composition and microbial diversity. These findings may have implications for healing outcomes and the risk of infection in the dental implant bed and should therefore be further explored

Evaluation of the Microbiological Performance and Potential Clinical Impact of New Rapid Molecular Assays for the Diagnosis of Bloodstream Infections

Bloodstream infection (BSI) is a critical medical emergency associated with a high mortality rate. Rapid and accurate identification of the causative pathogen and the results of antimicrobial susceptibility testing are crucial for initiating appropriate antimicrobial therapy. The aim of this study was to evaluate the performance of a new rapid PCR Molecular Mouse System (MMS) for the identification of Gram-negative bacteria (GNB) and GNB resistance genes directly from a positive blood culture (BC). The validation of these rapid multiplex assays was carried out in a real hospital setting. A total of 80 BSI episodes were included in our study and the results were compared with culture-based methods. BC samples in which GNB had previously been detected microscopically and which originated from different hospital wards were analysed. The MMS GNB identification assay achieved a sensitivity of 98.7% and a specificity of 100% for the covered pathogens. In one BC sample, Klebsiella aerogenes was identified at the family level (Enterobacteriaceae) with MMS. However, in three polymicrobial samples, MMS identified bacteria that were not detected by culture-based methods (Klebsiella pneumoniae, K. aerogenes and Stenotrophomonas maltophilia). MMS also showed excellent overall performance in the detection of GNB resistance markers (100% sensitivity and 100% specificity). The type of extended-spectrum beta-lactamase (ESBL) resistance gene identified correctly with MMS was CTX-M-1/9 (n = 17/20), alone or in combination with SHV-type β-lactamase or with the different types of carbapenemase genes. MMS detected one carbapenemase gene of each type (KPC, NDM and OXA-23) and six OXA-48 genes. In addition, the colistin resistance gene mcr-1 was detected in one positive BC with Escherichia coli (E. coli). The time to result was significantly shorter for MMS than for routine culture methods. A retrospective analysis of the patients’ medical records revealed that a change in empirical antimicrobial therapy would have been made in around half of the patients following the MMS results. These results support the use of MMS as a valuable complement to conventional culture methods for more rapid BSI diagnosis and adjustment of empirical therapy

Development and Validation of the CARRA-VID Prognostic Score: C Reactive Protein to Albumin Ratio, Red Blood Cell Distribution Width and Age-Based Score for Prognostication of Hospitalized COVID-19 Patients

Patients hospitalized due to coronavirus disease 2019 (COVID-19) usually present with severe or critical intensity of symptoms, accompanied by a marked systemic inflammatory response. Classical inflammatory biomarkers, C-reactive protein (CRP), albumin, and red blood cell distribution width (RDW) have previously been reported to be prognostic in hospitalized COVID-19 patients. We performed a retrospective analysis of two large cohorts (2305 and 2328 patients, respectively) of consecutive hospitalized COVID-19 patients with mostly severe and critical symptoms admitted to the tertiary referral center to develop and validate a prognostic score for 30-day mortality based on CRP-to-Albumin-Ratio (CAR), RDW, and age (termed CARRA-VID score). We identified 6 prognostic categories: very low, low, intermediate-1, intermediate-2, high, and very high risk, with corresponding 30-day mortality rates of 2.7%, 10.7%, 30.9%, 47.1%, 61.9%, and 89.7%, respectively. Effective risk stratification was validated in an independent cohort of patients and remained independent of the World Health Organization-defined disease severity and other commonly utilized risk scores. Additional analyses evaluated the score across different time periods dominated by distinct viral variants. We also present a simplified 3-tiered version of the score. A Microsoft Excel Workbook containing the score calculator is provided

APPLICATION OF MATHEMATICAL MODELLING IN THE ASSESSMENT OF SORPTION AND DESORPTION CAPACITY OF NEONICOTINOID INSECTICIDES IN AGRYCULTURAL SOILS Doctoral thesis

Ciljevi istraživanja: U Slavoniji se neonikotinoidi koriste kao učinkovita sredstva za zaštitu nasada šećerne repe. Njihovo ponašanje u tlu ovisi o karakteristikama tla, ali i o kemijskoj strukturi molekule. Cilj ovog istraživanja bio je ispitati utjecaj raznolikosti karakteristika tala i kemijske strukture neonikotinoida na intenzitet njihova vezivanja na koloide tla. Također, istražiti mogu li procesi sorpcije i desorpcije, te u kojoj mjeri, kontrolirati dostupnost insekticida i druge procese koji određuju njihovu sudbinu u okolišu tla. Metode: Uzorci tla prikupljeni su iz dvije hrvatske županije: Požeško-slavonske i Sisačko-moslavačke i okarakterizirani teksturno i fizikalno-kemijski. Kinetički i ravnotežni eksperimenti sorpcije i desorpcije acetamiprida, tiakloprida i imidakloprida provedeni su standardnom „šaržnom“ metodom, dok su njihovi ostatci analizirani pomoću HPLC-MS/MS. Za objašnjenje kinetike sorpcije i desorpcije korišteni su modeli: pseudo-prvog reda, Elovich-ev, unutar-čestične difuzije i bifazni model kinetike prvoga reda (TSM). Za ravnotežne procese korišteni su Freundlich-ov, Langmuir-ov i Temkin-ov model. Rezultati: Iako su analizirani neonikotinoidi slični u veličini i kemijskoj strukturi, rezultati su pokazali značajne razlike u kinetičkom i ravnotežnom ponašanju procesa sorpcije/desorpcije. TSM i Freundlich-ov model su se pokazali najprimjerenijima za objašnjenje tih procesa. Tiakloprid je pokazao bržu sorpciju u usporedbi s acetamipridom i imidaklopridom, a u svim tlima bio je jače sorbiran u ravnoteži. Unutarčestična difuzija bila je relevantan proces sorpcije acetamiprida i imidakloprida, ali ne i za tiakloprid. Acetamiprid je lakše desorbiran od preostala dva neonikotinoida. Acetamiprid i imidakloprid mogu se kategorizirati kao srednje pokretni insekticidi s manjom tendencijom sorpcije, što povećava vjerojatnost kontaminacije podzemnih voda, dok je tiakloprid slabo pokretan. Sadržaj organske tvari (OC), kao i njena priroda i struktura, imali su ključnu ulogu u sorpciji/desorpciji neonikotinoida. Aromatičnost je pogodovala sorpciji, dok je veća koncentracija hidrofilnih karboksilnih skupina potisnula sorpciju. Imidakloprid je podjednako ispunio sorpcijska mjesta u gumastoj i staklastoj fazi OC bez obzira na koncentraciju. Sorpcija tiakloprida pri niskim koncentracijama i acetamiprida pri visokim koncentracijama bila je kontrolirana hidrofilnim aromatskim strukturama, koje djeluju kao "hvatači" insekticida unutar pora staklaste faze OC. Razlike u ponašanju sorpcije/desorpcije proizlaze i iz varijacija u molekularnoj strukturi; tiazolidinski prsten koji sadrži sumpor u tiaklopridu uzrokuje razlike u topljivosti u vodi, lipofilnosti i kiselo-baznim svojstvima, a time i u interakcijama sa sastojcima tla. Preko atoma N u piridinskom prstenu, neonikotinoidi mogu stvarati π–π ili p–π interakcije donor-akceptor elektrona s aromatskim dijelovima organske tvari. Također, heteroatomi N, S i Cl mogu djelovati kao akceptori vodikovih (H–) veza i stvarati H–veze s funkcionalnim skupinama koje doniraju H u tlu. Zaključak: Matematički modeli pokazali su se kao učinkoviti alati za predviđanje pokretnosti neonikotinoida u tlu, pomažući u razjašnjenju mehanizama sorpcije/desorpcije i predviđanju njihove toksičnosti za ljudsku populaciju. Rezultati istraživanja doprinijet će razvoju budućih strategija sanacije i pomoći u tumačenju mogućih uzroka kontaminacije usjeva i ekosustava tla, čak i u slučajevima zabrane uporabe neonikotinoida.Objectives: In Slavonia, neonicotinoids are used as effective means to protect of sugar beet planatations. Their behavior in the soil depends on the soil characteristics and the chemical structure of the molecules. This research aimed to examine how variability in soil characteristics and the chemical structure of neonicotinoids affects their binding to soil colloids, and whether sorption and desorption processes influence the availability of these insecticides and other processes determining their fate in the soil environment. Methods: Soil samples were collected from Požega-Slavonia and Sisak-Moslavina counties, and characterized texturally and physico-chemically. Kinetic and equilibrium sorption and desorption experiments of acetamiprid, thiacloprid, and imidacloprid were carried out using the standard "batch" method, while their residues were analyzed using HPLC-MS/MS. Models used for kinetics analysis included pseudo-first order, Elovich’s, intra-particle diffusion, and first-order biphasic model (TSM), as well as Freundlich, Langmuir, and Temkin models for equilibrium processes. Results: Despite their similar size and chemical structure, the neonicotinoids exhibited differences in sorption/desorption behavior. The TSM and Freundlich models proved to be the most suitable for explaining these processes. Thiacloprid exhibited faster sorption compared to acetamiprid and imidacloprid, and in all soils it was more strongly sorbed at equilibrium. Intraparticle diffusion was a relevant sorption process for acetamiprid and imidacloprid, but not for thiacloprid. Acetamiprid was more easily desorbed, classifying it and imidacloprid as moderately mobile insecticides with a lower sorption tendency, which increases the likelihood of groundwater contamination, while thiacloprid was poorly mobile. Organic carbon content (OC) and its structure had a crucial role in the sorption/desorption of neonicotinoids. Aromaticity favored sorption, while a higher concentration of hydrophilic carboxyl groups suppressed sorption. Imidacloprid uniformly occupied sorption sites in both the rubbery and glassy phases of OC, regardless of concentration. The sorption of thiacloprid at low concentrations and acetamiprid at high concentrations was controlled by hydrophilic aromatic structures, which acted as "traps" for the insecticides within the pores of the glassy phase of OC. Differences in sorption/desorption behavior also results from variations in molecular structure; the sulfur-containing thiazolidine ring in thiacloprid causes differences in water solubility, lipophilicity, and acid-base properties, and thus in the interactions with soil constituents. Through the N atom in the pyridine ring, neonicotinoids can form π π or p–π electron donor-acceptor interactions with aromatic parts of organic matter. Additionally, heteroatoms N, S, and Cl can act as hydrogen (H–) bond acceptors, and form H–bonds with H-donating functional groups in the soil. Conclusion: Mathematical models have proven to be useful tools for predicting the mobility of neonicotinoids in soil, helping to elucidate sorption/desorption mechanisms and predict their toxicity to the human population. The findings of this research will contribute to the development of future remediation strategies and help in the interpretation of possible causes of crops and soil ecosystem contamination, even in cases of banning the use of neonicotinoids

Efficacy, Toxicity and Effect of Pretreatment Cardiologic Consultation on Outcomes of Ibrutinib Therapy for Chronic Lymphocytic Leukemia—A KroHem Study

Background/Objectives: Ibrutinib has revolutionized the treatment of chronic lymphocytic leukemia but has off-target side effects, most notably cardiac. In order to evaluate the efficacy and toxicity of ibrutinib treatment, risk factors for adverse outcomes and the influence of pretreatment cardiologic evaluation, KroHem collected data on Croatian patients with chronic lymphocytic leukemia treated with this drug. Methods: This is a retrospective survey performed in order to analyze the efficacy and toxicity of ibrutinib in a real-life setting. Patients starting therapy with ibrutinib for chronic lymphocytic leukemia between the time the drug became reimbursable in 2015 and 31 December 2021 were included, irrespective of treatment line. Results: We identified 436 patients fulfilling entry criteria; 404 (92.7%) responded to treatment. Cardiovascular side effects occurred in 25.0% of patients and hemorrhagic in 15.6%. The dose of ibrutinib was permanently reduced in 22.2% of patients. Median follow-up of the cohort was 29 months (IQR 18–41 months), estimated median overall survival 75 months (IQR 36 months–not reached), progression-free survival 54 months (IQR 24–81 months) and time on ibrutinib treatment 44 months (IQR 14–78 months). Factors significantly related to overall survival in multivariate analysis were stage, treatment line and age. Factors significantly related to progression-free survival in multivariate analysis were treatment line, age and pretreatment history or ECG finding of cardiac arrhythmia. Factors significantly related to time on ibrutinib treatment in multivariate analysis were age, pretreatment history or ECG finding of cardiac arrhythmia, and permanent dose reduction for toxicity. Sex, FISH and the presence of arterial hypertension were not independently significantly related to any of these outcomes. Pretreatment cardiologic consultation did not improve time on ibrutinib therapy, progression-free survival, overall survival, risk of stopping treatment due to cardiovascular side effects or risk of cardiovascular or sudden death, neither in the whole cohort nor in the subgroup of patients with and without pretreatment cardiac arrhythmia. Conclusions: Our analysis confirms the efficacy and tolerability of ibrutinib for the treatment of chronic lymphocytic leukemia. Patients older than 75 do significantly less well. Routine pretreatment cardiologic consultation does not improve outcomes and should not be considered part of standard pretreatment assessment without additional proof of its usefulness. Future investigations should aim at identifying predictive factors, mechanisms, and preventive strategies for reducing cardiotoxicity in chronic lymphocytic leukemia patients taking Bruton tyrosine kinase inhibitors

Treatment approaches for patients with multiple sclerosis during pregnancy and breastfeeding

Multipla skleroza (MS) autoimuna je neurodegenerativna bolest koja najčešće pogađa žene u reproduktivnoj dobi. Tradicionalno se trudnoća smatrala rizičnim razdobljem za žene s MS-om, no novija istraživanja pokazuju kako trudnoća u većini slučajeva ima povoljan učinak na tijek bolesti. Tijekom trudnoće dolazi do prirodne imunosupresije, što smanjuje rizik od relapsa. Nasuprot tome, postpartalno razdoblje karakterizira porast rizika od ponovnog aktiviranja bolesti. Liječenje MS-a u trudnoći i tijekom dojenja izazov je zbog ograničenih podataka o sigurnosti lijekova koji modificiraju tijek bolesti (DMT). Lijekovi kao što su interferon beta i glatiramer acetat imaju najviše podataka o sigurnosti, dok se visoko učinkoviti lijekovi poput natalizumaba i anti-CD20 protutijela mogu koristiti u žena s agresivnijim oblicima bolesti uz oprez i individualiziran pristup. Planiranje trudnoće kod žena s MS-om uključuje procjenu bolesti, prilagodbu terapije i pravovremenu edukaciju. Isključivo dojenje potencijalno smanjuje rizik od postpartalnih relapsa, iako su rezultati još uvijek nedovoljno jasni. Cilj ovog rada bio je prikazati trenutno dostupne spoznaje o liječenju žena s MS-om u trudnoći i dojenju te naglasiti važnost interdisciplinarne suradnje i potrebu za dodatnim istraživanjima u ovoj specifičnoj populaciji bolesnica.Multiple sclerosis (MS) is a chronic autoimmune disease that primarily affects women of reproductive age. While pregnancy was once discouraged in women with MS, current evidence suggests that pregnancy is generally safe and may even reduce disease activity due to natural immunosuppressive mechanisms. However, the postpartum period is associated with an increased risk of relapses. Managing MS during pregnancy and breastfeeding is complex due to limited safety data on disease-modifying therapies (DMTs). Interferon beta and glatiramer acetate have the most established safety profiles, while highly effective treatments such as natalizumab and anti-CD20 monoclonal antibodies may be considered for women with highly active disease, provided a case-by-case evaluation is done. Family planning, preconception counseling, and timely treatment adjustments are essential to reduce risks for both mother and child. Exclusive breastfeeding may provide some protective effect against postpartum relapses, though further research is needed to confirm this association. This thesis aimed to present current knowledge on MS treatment during pregnancy and lactation, emphasizing the importance of individualized care and the urgent need for more clinical research to inform treatment decisions in this population

Aspirin Versus LMWH for Thromboprophylaxis Following Hip or Knee Arthroplasty—Clinical Implications and Budget Impact

ABSTRACT Venous thromboembolism (VTE) remains a significant concern for patients undergoing hip or knee arthroplasty, with a need to balance effective thromboprophylaxis and bleeding risk. We aimed to compare the efficacy, safety, and budget impact of aspirin versus low‐molecular‐weight heparin (LMWH) as sole thromboprophylactic agents initiated immediately postoperatively in this population. First, we conducted a systematic review of randomized controlled trials (RCTs) from Ovid MEDLINE, Embase, and Cochrane CENTRAL databases, assessing clinical outcomes and healthcare costs. Subsequently, a simplified budget impact analysis was performed using data from the largest identified and most recent RCT (CRISTAL trial) and its secondary analyses. Primary outcomes included symptomatic VTE, bleeding events, and reoperation rates. Through a systematic search, seven RCTs were considered to be eligible, with the CRISTAL trial providing the most compelling evidence. Aspirin was non‐inferior to LMWH for all‐cause mortality but was associated with a significantly higher symptomatic VTE rate (3,27% vs. 1,76%) and deep vein thrombosis (DVT), predominantly distal DVT. The budget impact analysis revealed that despite aspirin's lower per tablet cost, thromboprophylaxis with LMWH led to annual savings of $35,912,459 to$110,431,241 for U.S. healthcare stakeholders, and $17,075 to$56,450 for single hospitals performing 1000 arthroplasty procedures annually. To conclude, enoxaparin appears to offer superior clinical efficacy and cost‐effectiveness compared to aspirin for thromboprophylaxis following hip and knee arthroplasty. These findings support the preferential use of LMWH in this setting, while highlighting the need for further investigation into the clinical significance of aspirin's higher distal DVT and pulmonary embolism risk

Integrated Chemical and Biological Evaluation of Linden Honeydew Honey from Bosnia and Herzegovina: Composition and Cellular Effects

Honeydew honey (HH) is a distinctive type of honey known for its dark colour, high mineral and polyphenol content, and pronounced biological activity. This study continues previous research on beech and chestnut honeydew honeys by presenting a comprehensive analysis of linden honeydew honey (LHH) from Bosnia and Herzegovina—a variety that, until now, has not been characterised in detail. Physicochemical parameters confirmed its classification as HH, with high electrical conductivity (1.21 mS/cm) and low moisture (15.1%). GC-MS analysis revealed a unique volatile profile dominated by α-terpinolene (17.4%), distinguishing LHH from other HH types. The sample exhibited high total phenolic content (816.38 mg GAE/kg) and moderate antioxidant capacity (1.11 mmol TE/kg). Antimicrobial testing demonstrated strong activity against Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA), with lower efficacy against Gram-negative bacteria. No cytotoxic effects were observed in HaCaT keratinocytes at concentrations up to 60 mg/mL, and wound healing assays showed improved scratch closure reaching approximately 30% after 24 h and 41% after 48 h compared to the control. These results indicate that LHH possesses promising bioactive properties and potential for dermatological application. Further studies with broader sample sets are needed to explore variability and confirm the therapeutic relevance of LHH in comparison to other honeydew types

Radiogenomics of Stereotactic Radiotherapy: Genetic Mechanisms Underlying Radiosensitivity, Resistance, and Immune Response

Stereotactic body radiotherapy (SBRT) delivers ablative radiation doses with sub-millimeter precision. Radiogenomic studies, meanwhile, provide insights into how tumor-intrinsic genetic factors influence responses to such high-dose treatments. This review explores the radiobiological mechanisms underpinning SBRT efficacy, emphasizing the roles of DNA damage response (DDR) pathways, tumor suppressor gene alterations, and inflammatory signaling in shaping tumor radiosensitivity or resistance. SBRT induces complex DNA double-strand breaks (DSBs) that robustly activate DDR signaling cascades, particularly via the ATM and ATR kinases. Tumors with proficient DNA repair capabilities often resist SBRT, whereas deficiencies in key repair genes can render them more susceptible to radiation-induced cytotoxicity. Mutations in tumor suppressor genes may impair p53-dependent apoptosis and disrupt cell cycle checkpoints, allowing malignant cells to evade radiation-induced cell death. Furthermore, SBRT provokes the release of pro-inflammatory cytokines and activates innate immune pathways, potentially leading to immunogenic cell death and reshaping the tumor microenvironment. Radiogenomic profiling has identified genomic alterations and molecular signatures associated with differential responses to SBRT and immune activation. These insights open avenues for precision radiotherapy approaches, including the use of genomic biomarkers for patient selection, the integration of SBRT with DDR inhibitors or immunotherapies, and the customization of treatment plans based on individual tumor genotypes and immune landscapes. Ultimately, these strategies aim to enhance SBRT efficacy and improve clinical outcomes through biologically tailored treatment. This review provides a comprehensive summary of current knowledge on the genetic determinants of response to stereotactic radiotherapy and discusses their implications for personalized cancer treatment

Lessons learned from RadiologyNET foundation models for transfer learning in medical radiology

Deep learning models require large amounts of annotated data, which are hard to obtain in the medical field, as the annotation process is laborious and depends on expert knowledge. This data scarcity hinders a model's ability to generalise effectively on unseen data, and recently, foundation models pretrained on large datasets have been proposed as a promising solution. RadiologyNET is a custom medical dataset that comprises 1,902,414 medical images covering various body parts and modalities of image acquisition. We used the RadiologyNET dataset to pretrain several popular architectures (ResNet18, ResNet34, ResNet50, VGG16, EfficientNetB3, EfficientNetB4, InceptionV3, DenseNet121, MobileNetV3Small and MobileNetV3Large). We compared the performance of ImageNet and RadiologyNET foundation models against training from randomly initialiased weights on several publicly available medical datasets: (i) Segmentation-LUng Nodule Analysis Challenge, (ii) Regression-RSNA Pediatric Bone Age Challenge, (iii) Binary classification-GRAZPEDWRI-DX and COVID-19 datasets, and (iv) Multiclass classification-Brain Tumor MRI dataset. Our results indicate that RadiologyNET-pretrained models generally perform similarly to ImageNet models, with some advantages in resource-limited settings. However, ImageNet-pretrained models showed competitive performance when fine-tuned on sufficient data. The impact of modality diversity on model performance was tested, with the results varying across tasks, highlighting the importance of aligning pretraining data with downstream applications. Based on our findings, we provide guidelines for using foundation models in medical applications and publicly release our RadiologyNET-pretrained models to support further research and development in the field. The models are available at https://github.com/AIlab-RITEH/RadiologyNET-TL-models