Trait-based sensitivity of large mammals to a catastrophic tropical cyclone.

Extreme weather events perturb ecosystems and increasingly threaten biodiversity1. Ecologists emphasize the need to forecast and mitigate the impacts of these events, which requires knowledge of how risk is distributed among species and environments. However, the scale and unpredictability of extreme events complicate risk assessment1-4-especially for large animals (megafauna), which are ecologically important and disproportionately threatened but are wide-ranging and difficult to monitor5. Traits such as body size, dispersal ability and habitat affiliation are hypothesized to determine the vulnerability of animals to natural hazards1,6,7. Yet it has rarely been possible to test these hypotheses or, more generally, to link the short-term and long-term ecological effects of weather-related disturbance8,9. Here we show how large herbivores and carnivores in Mozambique responded to Intense Tropical Cyclone Idai, the deadliest storm on record in Africa, across scales ranging from individual decisions in the hours after landfall to changes in community composition nearly 2 years later. Animals responded behaviourally to rising floodwaters by moving upslope and shifting their diets. Body size and habitat association independently predicted population-level impacts: five of the smallest and most lowland-affiliated herbivore species declined by an average of 28% in the 20 months after landfall, while four of the largest and most upland-affiliated species increased by an average of 26%. We attribute the sensitivity of small-bodied species to their limited mobility and physiological constraints, which restricted their ability to avoid the flood and endure subsequent reductions in the quantity and quality of food. Our results identify general traits that govern animal responses to severe weather, which may help to inform wildlife conservation in a volatile climate.

DNA-Targeted Complexes of Tc and Re for Biomedical Applications.

Due to their favorable chemical features, Re and Tc complexes have been widely used for the development of new therapeutic agents and imaging probes to solve problems of biomedical relevance. This review provides an update of the most relevant research efforts towards the development of novel cancer theranostic agents using Re and Tc-based compounds interacting with specific DNA structures. This includes a variety of homometallic complexes, namely those containing M(CO)3 (M=Re, Tc) moieties, that exhibit different modes of interaction with DNA, such as covalent binding, intercalation, groove binding or G-quadruplex DNA binding. Additionally, heterometallic complexes, designed to potentiate synergistic effects of different metal centers to improve DNA-targeting, cytotoxicity and fluorescence properties, are also reviewed. Particular attention is also given to 99m Tc- and 188 Re-labeled oligonucleotides that have been widely explored to develop imaging and therapeutic radiopharmaceuticals through the in vivo hybridization with a specific complementary DNA or RNA target sequence to provide useful molecular tools in precision medicine for cancer diagnosis and treatment. Finally, the need for further improvement of DNA-targeted Re and Tc-based compounds as potential therapeutic and diagnostic agents is highlighted, and future directions are discussed.

Targeting of G-quadruplex DNA with 99mTc(I)/Re(I) Tricarbonyl Complexes Carrying Pyridostatin Derivatives.

The main goal of this work was to elucidate the potential relevance of (radio)metal chelates of 99mTc and Re targeting G-quadruplex structures for the design of new tools for cancer theranostics. 99mTc provides the complexes with the ability to perform single-photon-emission computed tomography imaging studies, while the Re complexes should act as anticancer agents upon interaction with specific G4 DNA or RNA structures present in tumor tissues. Towards this goal, we have developed isostructural 99mTc(I) and Re(I) tricarbonyl complexes anchored by a pyrazolyl-diamine (Pz) chelator carrying a pendant pyridostatin (PDS) fragment as the G4-binding motif. The interaction of the PDF-Pz-Re (8) complex with different G4-forming oligonucleotides was studied by circular dichroism, fluorescence spectroscopy and FRET-melting assays. The results showed that the Re complex retained the ability to bind and stabilize G4-structures from different DNA or RNA sequences, namely those present on the SRC proto-oncogene and telomeric RNA (TERRA sequence). PDF-Pz-Re (8) showed low to moderate cytotoxicity in PC3 and MCF-7 cancer cell lines, as typically observed for G4-binders. Biodistribution studies of the congener PDF-Pz-99mTc (12) in normal mice showed that the complex undergoes a fast blood clearance with a predominant hepatobiliary excretion, pointing also for a high in vitro stability.

Synthesis and Preclinical Evaluation of PSMA-Targeted 111In-Radioconjugates Containing a Mitochondria-Tropic Triphenylphosphonium Carrier.

Nuclear DNA is the canonical target for biological damage induced by Auger electrons (AE) in the context of targeted radionuclide therapy (TRT) of cancer, but other subcellular components might also be relevant for this purpose, such as the energized mitochondria of tumor cells. Having this in mind, we have synthesized novel DOTA-based chelators carrying a prostate-specific membrane antigen (PSMA) inhibitor and a triphenyl phosphonium (TPP) group that were used to obtain dual-targeted 111In-radioconjugates ([111In]In-TPP-DOTAGA-PSMA and [111In]In-TPP-DOTAGA-G3-PSMA), aiming to promote a selective uptake of an AE-emitter radiometal (111In) by PSMA+ prostate cancer (PCa) cells and an enhanced accumulation in the mitochondria. These dual-targeted 111In-radiocomplexes are highly stable under physiological conditions and in cell culture media. The complexes showed relatively similar binding affinities toward the PSMA compared to the reference tracer [111In]In-PSMA-617, in line with their high cellular uptake and internalization in PSMA+ PCa cells. The complexes compromised cell survival in a dose-dependent manner and in the case of [111In]In-TPP-DOTAGA-G3-PSMA to a higher extent than observed for the single-targeted congener [111In]In-PSMA-617. µSPECT imaging studies in PSMA+ PCa xenografts showed that the TPP pharmacophore did not interfere with the excellent in vivo tumor uptake of the "golden standard" [111In]In-PSMA-617, although it led to a higher kidney retention. Such kidney retention does not necessarily compromise their usefulness as radiotherapeutics due to the short tissue range of the Auger/conversion electrons emitted by 111In. Overall, our results provide valuable insights into the potential use of mitochondrial targeting by PSMA-based radiocomplexes for efficient use of AE-emitting radionuclides in TRT, giving impetus to extend the studies to other AE-emitting trivalent radiometals (e.g., 161Tb or 165Er) and to further optimize the designed dual-targeting constructs.

Epidemiological profile of malaria in a rural community in the Amazon, Mato Grosso State, Brazil, 2011.

BACKGROUND: More than 95% of malaria transmission in Brazil occurs in the Legal Amazon Region, which in 2010 recorded around 333,429 cases reported in the Epidemiological Surveillance Information System-Malaria (Sivep_malaria), presenting an annual parasitic incidence (IPA) of 13.1 cases/1000 inhabitants. METHODS: This was a descriptive study that measured the community prevalence of Plasmodium infection and its relationship with land use in Três Fronteiras District, Colniza Municipality, Mato Grosso State. Data were collected during household visits in July 2011, with blood collection from finger pricks for the preparation of thick smear slides, and completion of a standardized case notification form. A georeferenced database was analysed, with land use evaluated as categorical variables. A kernel density map was built to show the density of cases and their location. RESULTS: Of the 621 respondents, 68(11%) had Plasmodium infection: 39 (57.4%) with Plasmodium vivax, 27(39.7%) with Plasmodium falciparum and two (2.9%) with mixed infections. Among infected individuals, 49 (72.1%) were men. Cases of malaria were distributed over the district, with greater occurrence of cases per household in open areas close to the mining company and artisanal mining sites. The was a greater density of cases located in the gold mining region. CONCLUSION: Transmission of malaria in Três Fronteiras District has a heterogeneous distribution. Individuals residing in mining and timber extraction sites have increased occurrence of Plasmodium infection.

Developmental Defects of Enamel: A Bibliometric Analysis of the Top 100 Most-Cited Papers.

BACKGROUND: Developmental defects of enamel (DDE) are a result of disturbances during formation and maturation of the enamel. Evaluating the most-cited DDE papers can provide important tools that point to the gaps and strengths of this important topic in dentistry. SUMMARY: This bibliometric study analyzed the 100 most-cited papers on DDE. Using a combined keyword search strategy, the 100 most-cited papers were selected in the Web of Science Core Collection. Papers that addressed any type of DDE were included. The extracted data were title, number of citations, study theme, authorship, journal, type of DDE, type of dentition (primary or permanent), type of diagnosis, study design, year, and country of publication. The bibliometric networks were generated through VOSviewer software. The 100 papers had a range from 78 to 459 citations. The main themes of studies were etiopathogenesis (53%), prevalence and incidence (22%), and diagnosis (8%). The authors with the highest number of citations were Goodman AH and Rose JC (459 citations). Most articles were published in dental journals (47%). The most studied types of DDE were fluorosis and amelogenesis imperfecta in the permanent dentition (47%). Observational (24%) and non-systematic reviews (24%) were the most common study designs and ranged from 1977 to 2019. The country with the highest number of publications was the USA (41%). KEY MESSAGES: Most of the top 100 DDE papers were about fluorosis and amelogenesis imperfecta, with top papers from three continents with English as the native language. This topic is of great importance in dentistry, and the need for further studies is highlighted, especially regarding the diagnosis and treatment of some DDEs.

Passive smoking and early childhood caries: when adult addiction affects children's oral health.

DATA SOURCES: Medline via EBSCOhost, PubMed, and Scopus up to February 2023. STUDY SELECTION: Observational studies in the English language published in peer-reviewed journals, encompassing children aged 0 to 6 years exposed to environmental tobacco smoke and passive smoking, with a focus on oral health conditions in deciduous dentition. DATA EXTRACTION AND SYNTHESIS: Data were extracted from 25 papers, encompassing study details, oral health, and smoking exposure. Confounding factors such as parental education, income, diet, fluoride exposure, gender, and oral hygiene practices were also identified. The risk of bias was assessed using the Newcastle-Ottawa Scale (NOS). Frequency effect size was calculated for health variables and smoking exposure. RESULTS: Most studies identified a correlation between passive smoking and an increased prevalence of dental caries. Environmental tobacco smoke exposure and dental caries risk were influenced by factors such as low parental education, socioeconomic status, dietary habits, oral hygiene, and gender. CONCLUSIONS: The results of this systematic review strongly indicate a significant association between dental caries in deciduous dentition and passive smoking.

Structural Perspective into the Interaction of an Oncogenesis-Relevant pre-miRNA G-Quadruplex Ligand Carrier with the Protein Nucleolin.

The structural determinants of the interaction of the G-quadruplex (G4) motif found in precursor miRNA 149 (rG4) with the acridine orange derivative C8 , a G4 ligand stabilizer possessing anticancer activity, and the protein nucleolin (overexpressed in cancer cells) were investigated by Nuclear Magnetic Resonance (NMR) spectroscopy. For the rG4/C8 complex, the results revealed a strong stabilizing interaction between the aromatic core and the iodinated ring of the C8 ligand with the rG4 structure. The NMR study revealed also different interaction patterns between nucleolin and rG4 and nucleolin and rG4/C8 complex. In the absence of the ligand, rG4 establishes interactions with polar residues of the protein while for the rG4/C8 complex, these contacts are mainly established with amino acids that have hydrophobic side chains. However, nucleolin chemical shift perturbation studies in the presence of rG4 or rG4/C8 reveal the same location between domains 1 and 2 of the protein, which suggests that the rG4 and rG4/C8 complex bind in this region. This puzzling structural study opens a new framework to study rG4/ligand/nucleolin complexes that might impact the biogenesis of miRNA 149.

rJararacin, a recombinant disintegrin from Bothrops jararaca venom: Exploring its effects on hemostasis and thrombosis.

Integrins are a family of heterodimeric transmembrane receptors which link the extracellular matrix to the cell cytoskeleton. These receptors play a role in many cellular processes: adhesion, proliferation, migration, apoptosis, and platelet aggregation, thus modulating a wide range of scenarios in health and disease. Therefore, integrins have been the target of new antithrombotic drugs. Disintegrins from snake venoms are recognized by the ability to modulate the activity of integrins, such as integrin αIIbß3, a fundamental platelet glycoprotein, and αvß3 expressed on tumor cells. For this reason, disintegrins are unique and potential tools for examining integrin-matrix interaction and the development of novel antithrombotic agents. The present study aims to obtain the recombinant form of jararacin and evaluate the secondary structure and its effects on hemostasis and thrombosis. rJararacin was expressed in the Pichia pastoris (P. pastoris) expression system and purified the recombinant protein with a yield of 40 mg/L of culture. The molecular mass (7722 Da) and internal sequence were confirmed by mass spectrometry. Structure and folding analysis were obtained by Circular Dichroism and 1H Nuclear Magnetic Resonance spectra. Disintegrin structure reveals properly folded with the presence of ß-sheet structure. rJararacin significantly demonstrated inhibition of the adhesion of B16F10 cells and platelets to the fibronectin matrix under static conditions. rJararacin inhibited platelet aggregation induced by ADP (IC50 95 nM), collagen (IC50 57 nM), and thrombin (IC50 22 nM) in a dose-dependent manner. This disintegrin also inhibited 81% and 94% of the adhesion of platelets to fibrinogen and collagen under continuous flow, respectively. In addition, rjararacin efficaciously prevents platelet aggregation in vitro and ex vivo with rat platelets and thrombus occlusion at an effective dose (5 mg/kg). The data here provides evidence that rjararacin possesses the potential as an αIIbß3 antagonist, capable of preventing arterial thrombosis.

A Bibliometric Analysis of the Top 100 Most-Cited Papers Concerning Dental Fluorosis.

A high number of citations can indicate the potential of any specific paper to influence other research and generate changes in clinical practice. Analyzing the most-cited papers in a certain scientific field may assist researchers to identify the influential papers as well their main characteristics. The present study aimed to analyze the 100 most-cited papers concerning dental fluorosis (DF) through a bibliometric review. A search was performed in the Web of Science Core Collection (WoS-CC) database in November 2021. The papers were displayed in descending order according to the number of citations in WoS-CC. Two independent researchers performed the selection. Scopus and Google Scholar were used to compare the number of citations with WoS-CC. The following data were extracted from the papers: title, authors, number and density of citations, institution, country, continent, year of publication, journal title, keywords, study design, and theme. Collaborative networks were generated using the VOSviewer software. The top 100 most-cited papers were published between 1974 and 2014 and were cited 6,717 times (ranging from 35 to 417). Community Dentistry and Oral Epidemiology (24%), Journal of Dental Research (21%), Journal of Public Health Dentistry (17%), and Caries Research (13%) published the most papers. Observational studies (60%) and literature reviews (19%) were the most common study designs. The main topics were epidemiology (44%) and fluoride intake (32%). The countries with the highest number of papers were the USA (44%), Canada (10%), and Brazil (9%). The University of Iowa (USA) had the most papers (12%). Levy SM was the author with the highest number of papers (12%). The 100 most-cited papers on DF were mainly observational studies focused on epidemiology and originated in North America. There were few interventional studies and systematic reviews among the most-cited papers concerning this topic.

Graphene Oxide Thin Films for Detection and Quantification of Industrially Relevant Alcohols and Acetic Acid.

Industrial environments are frequently composed of potentially toxic and hazardous compounds. Volatile organic compounds (VOCs) are one of the most concerning categories of analytes commonly existent in the indoor air of factories' facilities. The sources of VOCs in the industrial context are abundant and a vast range of human health conditions and pathologies are known to be caused by both short- and long-term exposures. Hence, accurate and rapid detection, identification, and quantification of VOCs in industrial environments are mandatory issues. This work demonstrates that graphene oxide (GO) thin films can be used to distinguish acetic acid, ethanol, isopropanol, and methanol, major analytes for the field of industrial air quality, using the electronic nose concept based on impedance spectra measurements. The data were treated by principal component analysis. The sensor consists of polyethyleneimine (PEI) and GO layer-by-layer films deposited on ceramic supports coated with gold interdigitated electrodes. The electrical characterization of this sensor in the presence of the VOCs allows the identification of acetic acid in the concentration range from 24 to 120 ppm, and of ethanol, isopropanol, and methanol in a concentration range from 18 to 90 ppm, respectively. Moreover, the results allows the quantification of acetic acid, ethanol, and isopropanol concentrations with sensitivity values of (3.03±0.12)∗104, (-1.15±0.19)∗104, and (-1.1±0.50)∗104 mL-1, respectively. The resolution of this sensor to detect the different analytes is lower than 0.04 ppm, which means it is an interesting sensor for use as an electronic nose for the detection of VOCs.

The State of the Art on Graphene-Based Sensors for Human Health Monitoring through Breath Biomarkers.

The field of organic-borne biomarkers has been gaining relevance due to its suitability for diagnosing pathologies and health conditions in a rapid, accurate, non-invasive, painless and low-cost way. Due to the lack of analytical techniques with features capable of analysing such a complex matrix as the human breath, the academic community has focused on developing electronic noses based on arrays of gas sensors. These sensors are assembled considering the excitability, sensitivity and sensing capacities of a specific nanocomposite, graphene. In this way, graphene-based sensors can be employed for a vast range of applications that vary from environmental to medical applications. This review work aims to gather the most relevant published papers under the scope of "Graphene sensors" and "Biomarkers" in order to assess the state of the art in the field of graphene sensors for the purposes of biomarker identification. During the bibliographic search, a total of six pathologies were identified as the focus of the work. They were lung cancer, gastric cancer, chronic kidney diseases, respiratory diseases that involve inflammatory processes of the airways, like asthma and chronic obstructive pulmonary disease, sleep apnoea and diabetes. The achieved results, current development of the sensing sensors, and main limitations or challenges of the field of graphene sensors are discussed throughout the paper, as well as the features of the experiments addressed.

Which dental and psychosocial factors may predict bullying occurrence in adolescence?

DESIGN: Cohort study. STUDY POPULATION: 639 preschool children at baseline (T1) and 429 adolescents at follow-up (T2). OUTCOME MEASURES: Oral health conditions (untreated dental caries, dental trauma and malocclusion, sense of coherence (SOC), self-perception of dental treatment needs, general and dental bullying. DATA ANALYSIS: Chi-square test, t-test, structural equation modeling, goodness of fit. RESULTS: Untreated dental caries and low socioeconomic status impacted the occurrence of both general and dental bullying. SOC and self-perceived dental treatment need were also important factors in the occurrence of bullying. CONCLUSIONS: The oral health condition that most impacted bullying episodes both directly and indirectly was untreated dental caries. Socioeconomic status was also an important cumulative predictor of the occurrence of episodes of bullying.

Retention in fissure resin-based sealants in schoolchildren: the etching step importance.

DESIGN: Randomized controlled trial. STUDY POPULATION: Forty-seven schoolchildren aged 9-10 years with fully sound/noncavitated erupted first permanent molars were included and randomly allocated in control and experimental groups in a split-mouth design. CASES: 47 schoolchildren (94 molars) fissure sealants applied using self-etch universal adhesive system. CONTROLS: 47 schoolchildren (94 molars) fissure sealants applied using conventional acid-etching technique. OUTCOME MEASURE: Retention of sealants and secondary caries incidence (ICDAS). DATA ANALYSIS: Chi-square test. RESULTS: Conventional acid-etch sealants retention was superior to self-etch after 6 and 24 months (p < 0.001), but no differences were found in caries incidence after 6 and 24 months (p > 0.05). CONCLUSIONS: Clinical retention of fissure sealants using conventional acid-etch technique is greater than self-etch.

Altered heart cytokine profile and action potential modulation in cardiomyocytes from Mas-deficient mice.

The renin-angiotensin system (RAS) is a key hormonal system. In recent years, the functional analysis of the novel axis of the RAS (ACE2/Ang-(1-7)/Mas receptor) revealed that its activation can become protective against several pathologies, including cardiovascular diseases. Mas knockout mice (Mas-KO) represent an important tool for new investigations. Indeed, extensive biological research has focused on investigating the functional implications of Mas receptor deletion. However, although the Mas receptor was identified in neonatal cardiomyocytes and also in adult ventricular myocytes, only few reports have explored the Ang-(1-7)/Mas signaling directly in cardiomyocytes to date. This study investigated the implication of Mas receptor knockout to the cytokine profile, energy metabolism, and electrical properties of mice-isolated cardiomyocytes. Here, we demonstrated that Mas-KO mice have modulation in some cytokines, such as G-CSF, IL-6, IL-10, and VEGF in the left ventricle. This model also presents increased mitochondrial number in cardiomyocytes and a reduction in the myocyte diameter. Finally, Mas-KO cardiomyocytes have altered action potential modulation after diazoxide challenge. Such electrical finding was different from the data showed for the TGR(A1-7)3292 (TGR) model, which overexpresses Ang-(1-7) in the plasma by 4.5, used by us as a control. Collectively, our findings exemplify the importance of understanding the ACE2/Ang-(1-7)/Mas pathway in cardiomyocytes and heart tissue. The Mas-KO mice model can be considered an important tool for new RAS investigations.

Optimizing the Decellularized Porcine Liver Scaffold Protocol.

There are few existing methods for shortening the decellularization period for a human-sized whole-liver scaffold. Here, we describe a protocol that enables effective decellularization of the liver obtained from pigs weigh 120 ± 4.2 kg within 72 h. Porcine livers (approx. 1.5 kg) were decellularized for 3 days using a combination of chemical and enzymatic decellularization agents. After trypsin, sodium deoxycholate, and Triton X-100 perfusion, the porcine livers were completely translucent. Our protocol was efficient to promote cell removal, the preservation of extracellular matrix (ECM) components, and vascular tree integrity. In conclusion, our protocol is efficient to promote human-sized whole-liver scaffold decellularization and thus useful to generate bioengineered livers to overcome the shortage of organs.

The Top 100 Most-Cited Papers in Erosive Tooth Wear: A Bibliometric Analysis.

Citation rates can be used as an indicator of the influence and relevance of scientific papers. The present study analyzed the 100 most-cited articles related to erosive tooth wear (ETW). The top 100 most-cited papers focusing on ETW topics were collected from the Web of Science database on November 11, 2020. The following bibliometric data were extracted from papers: title, authorship, institutions, countries, number of citations, year of publication, journal title, study design, topic, and keywords. Scopus and Google Scholar were searched to compare the number of citations. The VOSviewer software was used to generate collaborative network maps for the authors and keywords. The number of citations of the 100 most-cited papers ranged from 71 to 330 (average: 97.44). The papers were published between 1949 and 2015. Caries Research (28%) and The Journal of Dentistry (16%) presented the majority of papers. Laboratorial (44%) and observational studies (33%) were the most common study designs. The most studied topics were epidemiology (31%) and the erosive potential of substances (drinks, foods, or medicines) (22%). The countries with the highest number of most-cited papers were England (32%), Germany (18%), and Switzerland (10%). The University of Bern was the institution with the most papers (10%). Lussi A was the author with the highest number of papers in the top 100 (14%). The most common keyword was "dental erosion" with 64 occurrences. The top 100 most-cited papers related to ETW were composed mainly of laboratorial and observational studies focusing on epidemiology and the erosive potential of substances.

Prevalence of toothache and associated factors in children and adolescents: a systematic review and meta-analysis.

OBJECTIVES: This systematic review aimed to determine the pooled prevalence of toothache in children and adolescents and to verify its association with sociodemographic characteristics and dental caries experience. MATERIAL AND METHODS: Six main and three grey literature databases were searched in February 2021. Observational studies that assessed the prevalence of toothache and associated factors were included. The methodological quality of the studies was evaluated following the guidelines by Fowkes and Fulton. Proportion and association meta-analyses were performed in MedCalc and Revman software, respectively. The certainty of the evidence was determined with the GRADE approach. RESULTS: Seventy-one studies met the inclusion criteria. Thirty-two presented high methodological quality and a low risk of bias. The overall pooled prevalence of toothache in children and adolescents was 36.2% (95%CI: 33.0-39.42; I2: 99.72%; P < 0.001). Females (OR: 1.17; 95%CI: 1.08-1.26; I2: 91%; P < 0.001) and children and adolescents whose caregivers had ≤ 8 years of schooling (OR: 1.42; 95%CI: 1.30-1.56; I2: 77%; P < 0.001) presented the higher chance of reporting toothache. Dental caries experience increased the chance of reporting toothache in 3.49 times (95%CI: 2.70-4.51; I2: 92%; P < 0.001). The certainty of the evidence was very low. CONCLUSIONS: The overall pooled prevalence of toothache was considered high. Sociodemographic characteristics (sex and caregivers' educational level) and previous dental caries experience were significantly associated with toothache reports. CLINICAL RELEVANCE: Toothache is still a recurrent issue in many countries. Thus, it is essential to understand its possible associated factors to reduce the occurrence of new episodes.

Radiolabeled Gold Nanoseeds Decorated with Substance P Peptides: Synthesis, Characterization and In Vitro Evaluation in Glioblastoma Cellular Models.

Despite some progress, the overall survival of patients with glioblastoma (GBM) remains extremely poor. In this context, there is a pressing need to develop innovative therapy strategies for GBM, namely those based on nanomedicine approaches. Towards this goal, we have focused on nanoparticles (AuNP-SP and AuNP-SPTyr8) with a small gold core (ca. 4 nm), carrying DOTA chelators and substance P (SP) peptides. These new SP-containing AuNPs were characterized by a variety of analytical techniques, including TEM and DLS measurements and UV-vis and CD spectroscopy, which proved their high in vitro stability and poor tendency to interact with plasma proteins. Their labeling with diagnostic and therapeutic radionuclides was efficiently performed by DOTA complexation with the trivalent radiometals 67Ga and 177Lu or by electrophilic radioiodination with 125I of the tyrosyl residue in AuNP-SPTyr8. Cellular studies of the resulting radiolabeled AuNPs in NKR1-positive GBM cells (U87, T98G and U373) have shown that the presence of the SP peptides has a crucial and positive impact on their internalization by the tumor cells. Consistently, 177Lu-AuNP-SPTyr8 showed more pronounced radiobiological effects in U373 cells when compared with the non-targeted congener 177Lu-AuNP-TDOTA, as assessed by cell viability and clonogenic assays and corroborated by Monte Carlo microdosimetry simulations.

Searching for a Paradigm Shift in Auger-Electron Cancer Therapy with Tumor-Specific Radiopeptides Targeting the Mitochondria and/or the Cell Nucleus.

Although 99mTc is not an ideal Auger electron (AE) emitter for Targeted Radionuclide Therapy (TRT) due to its relatively low Auger electron yield, it can be considered a readily available "model" radionuclide useful to validate the design of new classes of AE-emitting radioconjugates. With this in mind, we performed a detailed study of the radiobiological effects and mechanisms of cell death induced by the dual-targeted radioconjugates 99mTc-TPP-BBN and 99mTc-AO-BBN (TPP = triphenylphosphonium; AO = acridine orange; BBN = bombesin derivative) in human prostate cancer PC3 cells. 99mTc-TPP-BBN and 99mTc-AO-BBN caused a remarkably high reduction of the survival of PC3 cells when compared with the single-targeted congener 99mTc-BBN, leading to an augmented formation of γH2AX foci and micronuclei. 99mTc-TPP-BBN also caused a reduction of the mtDNA copy number, although it enhanced the ATP production by PC3 cells. These differences can be attributed to the augmented uptake of 99mTc-TPP-BBN in the mitochondria and enhanced uptake of 99mTc-AO-BBN in the nucleus, allowing the irradiation of these radiosensitive organelles with the short path-length AEs emitted by 99mTc. In particular, the results obtained for 99mTc-TPP-BBN reinforce the relevance of targeting the mitochondria to promote stronger radiobiological effects by AE-emitting radioconjugates.