High HHV-8 viral load (VL) in Kaposi Sarcoma (KS) has been associated with Severe Immune Reconstitution Inflammatory Syndrome (Severe-IRIS-KS), which can occur after initiating cART, and leads to high mortality, particularly in patients with pulmonary involvement. We investigate if valganciclovir (as an anti-HHV-8 agent) initiated before cART reduces the mortality associated with Severe-IRIS-KS and the incidence of Severe-IRIS-KS.Open-label parallel-group randomized clinical trial in AIDS cART naïve patients with disseminated KS (DKS) as defined by at least two of the following: pulmonary, lymph-node, or gastrointestinal involvement, lymphedema, or ≥30 skin lesions. In the experimental group (EG), patients received valganciclovir 900 mg BID four weeks before cART and continued until week 48; in the control group (CG), cART was initiated on week 0. Non-severe-IRIS-KS was defined as: an increase in the number of lesions plus a decrease of ≥one log10 HIV-VL, or an increase of ≥50cells/mm3 or ≥2-fold in baseline CD4+cells. Severe-IRIS-KS was defined as abrupt clinical worsening of KS lesions and/or fever after ruling out another infection following cART initiation, and at least three of the following: thrombocytopenia, anemia, hyponatremia, or hypoalbuminemia.40 patients were randomized and 37 completed the study. In the ITT analysis, at 48 weeks, total mortality was the same in both groups (3/20), severe-IRIS-KS attributable mortality was 0/20 in the EG, compared with 3/20 in the CG (p = 0.09), similar to the per-protocol analysis: 0/18 in the EG, and 3/19 in the control group (p = 0.09). The crude incidence rate of severe-IRIS-KS was four patients developed a total of 12 episodes of Severe-IRIS-KS in the CG and two patients developed one episode each in the EG. Mortality in patients with pulmonary KS was nil in the EG (0/5) compared with 3/4 in the CG (P = 0.048). No difference was found between groups in the number of non-S-IRIS-KS events. Among survivors at week 48, 82% achieved >80% remission.Although mortality attributable to KS was lower in the EG the difference was not statistically significant. Pneumocystis jirovecci pneumonia heralded the beginning of the HIV/AIDS pandemic [Kaposi Sarcoma (KS) and pandemic ,2. Beforpandemic ,4 which pandemic ,6. The ipandemic –10. The pandemic –13 The cpandemic  which hiin vitro and in vivo activity against HHV-8 [High HHV-8 VL levels are associated with disease severity and mortality ,15 and, st HHV-8 –19 couldst HHV-8 ,21. A clst HHV-8 .We performed a randomized clinical trial to test the hypothesis that valganciclovir  started four weeks before cART initiation, in patients with disseminated Kaposi sarcoma (DKS), would reduce the mortality associated to Severe-IRIS-KS and the incidence of Severe-IRIS-KS events. We also evaluated whether or not in the presence of coinfections serum cytokine levels  and C-reactive Protein (CRP) correlated with Severe-IRIS-KS occurrence.The protocol was approved by the Comité de Ética en Investigación  (015/031/INI) (CEI/950/15) under the Declaration of Helsinki and US Federal Policy for the Protection of Human Subjects and registered at NIH Clinical Trails ID NCT03296553. All participants signed written informed consent.The protocol was approved by the institutional IRB on July 2015, and we recruited the first patient on October 2015. The translated protocol was uploaded to Clinical Trials on March 2016, and learned one year later than submission to Clinical Trials was incomplete, so the official date for registration to clinical trials was September 2017.1I1S1 Poor Prognosis according to the European Consensus-Based Interdisciplinary Guideline [We conducted an open-label parallel-group randomized clinical trial of patients with DKS  endoscopy with biopsy of lesions. Colonoscopy was performed only on patients with diarrhea or lower-GIT bleeding. Biopsies of enlarged lymph nodes were processed for culture and histopathological analysis. If indicated, we performed bronchoscopy with bronchoalveolar lavage (BAL), thoracic Gallium Scan, or PET-FDG scan.Candidates had an initial thorough clinical evaluation including a work-up to diagnose coinfections and rule out other neoplasms that comprised: ophthalmologic evaluation, computed tomography (CT) scan , bone marrow culture with bone biopsy, Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) serology, venereal disease research laboratory (VDRL) test and if indicated, a lumbar puncture to rule-out neurosyphilis, Histoplasma urinary antigen and serology, GenXpert MTB/RIF test Patients >18 years old, HIV+ naïve to cART with DKS, able and willing to provide written informed consent. DKS was defined as the presence of KS pulmonary disease and/or ≥30 KS skin lesions, with or without lymphedema, and/or lymph node involvement, and/or GIT KS involvement (biopsy proven at least in one site).Another concomitant malignancy, Multicentric Castleman Disease (MCD), steroid treatment two months before screening, active HBV or HCV or CMV end-organ disease, or severely ill patients with APACHE II score >15.Randomization. Patients were randomized by blocks of ten; the assigned group was written in closed envelopes: either to the Control Group (CG) to start cART immediately according to current Mexican Guidelines [idelines , or to tPatients who at the time of diagnosis presented with extensive KS involvement or bulky oropharyngeal lesions were treated with vincristine 2 mg  plus bleomycin 15 UI before cART, or at follow-up in case of S-IRIS-KS or persistence of extensive KS lesions.Visits were scheduled at baseline, and weeks 1, 2, 4, 8, 12, 16, 24, and 48. At each visit, an Infectious Diseases specialist performed a clinical evaluation and skin lesions were photographed. Blood samples were obtained for: WBC count, complete blood chemistry, urine analysis, CRP, D-Dimer, HIV VL with Abbott Real-time, HHV-8, CMV, and Epstein-Barr Virus (EBV) VL ELITe MGB KIT by ELITe InGenius Software, CD4+, and CD8+ cells count and percentage, . CD4/CD8 ratio and plasma levels of interleukin 6, 10 , tumor necrosis factor (TNF), and interferon-gamma (IFN-ɤ) were measured using a sandwich-type immunoassay, ELISA (Biolegend). Serology for syphilis, HBV, and HCV was repeated at weeks 24 and 48. If patients had an exacerbation of KS outside the scheduled visit they were re-evaluated with laboratory tests and image studies.3 or ≥2-fold from baseline CD4+ cells [For this trial, non-severe-IRIS-KS was considered in patients with an increase in the number of KS lesions plus a decrease of ≥one log10 of HIV-1 RNA VL or an increase of ≥50 cells/mm4+ cells . Severe-Clinical criteria: 1) Fever (without identified concomitant infection), 2) increase in the size or number of KS lesions, 3) exacerbation of lymphedema, 4) appearance or increase of otherwise unexplained lung opacities on the chest images with a negative Gallium-Scan and 4) appearance or increase of volume of pleural effusion.Laboratory criteria: 1) Thrombocytopenia <100,000 platelets/ml; 2) Anemia (decrease of at least 1 g/dl from previous measure with no obvious bleeding), 3) Hyponatremia <135 mEq/L and 4) Hypoalbuminemia <3.5 g/dL. Patients with severe IRIS KS can respond to bleomycin/vincristine therapy. We defined the resolution of a Severe-IRIS-KS: when platelets normalized, Hb increased (not due to a blood transfusion), albumin and sodium improved, and fever resolved.The main outcome was KS-attributable mortality at 48 weeks; secondary outcomes were the incidence of severe-IRIS-KS events, that is the number of patients with a least one severe IRIS-KS event. We also describe in an exploratory way the overall mortality, the mortality in patients with Pulmonary KS, and the total number of IRIS-KS events in the EG and the CG as we observed patients with more than one severe IRIS-KS episode.Improvement of KS lesions and KS remission were assessed by comparing the size and number of KS lesions from baseline to 48-week visit comparing pictures taken at every visit, and lung CT-scan for cases with pulmonary involvement.The sample size was calculated for a study power of 80% and an alpha of 0.05, with an estimated event rate (Severe-IRIS-KS-associated deaths) in the control group of 40%, and in the treated group of 5%. The sample number estimated in each group was 19, for a total of 38 patients.Descriptive statistics were performed, including the number of deaths, and the number of participants with Severe-IRIS-KS, in the CG and EG groups. Comparisons between EG and CG were performed as intention-to-treat (ITT), and as per-protocol analysis, for the primary outcome of mortality attributed to Severe-IRIS-KS by a Fisher´s exact test, and also comparing survival curves by Log-rank test. We also compared the EG and the CG groups by a Fisher´s exact test on the incidence of Severe-IRIS-KS, on overall mortality, and mortality in patients with pulmonary KS. During the study, we observed that some individuals had more than one Severe-IRIS-KS episode during the 48 weeks of follow-up, so we also compared the total number of Severe-IRIS-KS episodes in each group.The impact of treatment and time on IL-6, IL10, HHV-8 viral load, and C reactive protein were evaluated by fitting linear mixed effect models, and the impact of some covariates  on outcomes by fitting multivariate Poisson models. Analyses were performed using Stata Statistical program.A total of 95 male patients were screened, 40 of them were randomized  between The main characteristics of the experimental and control groups are depicted in Thirty-five (87%) patients had HHV-8 viremia at baseline alone or in combination with EBV or CMV (19 EG and 16 CG). Nine (22.5%) patients had pulmonary KS involvement four (20%) in the CG group and five (25%) in the EG. Eleven patients 27.5%) were diagnosed with HIV-polymorphic lymphoproliferative disorder [.5% were 1N1 influenza pneumonia see . In the neumonia .In the per-protocol analysis, we included 37 patients, 19 in the CG (one patient abandoned the protocol on week 12) and 18 in the EG , Fisher´s exact test) as depicted in p = 0.04).Four patients developed 12 episodes of Severe-IRIS-KS in the CG Incidence Rate (IR) (0.21 per 100 patient-observation days) and two patients developed one episode each in the EG , IL6  and CRP at baseline  increased the risk to develop Severe-IRIS-KS and valganciclovir use significantly decreased the risk of Severe-IRIS-KS occurrence  .We performed a linear mixed effect model analysis for longitudinal data for 4 variables IL-6, IL10, HHV-8 viral load, and CRP; the impact of treatment was not significant, at the p<0.05 level and with time, only CRP decreased significantly (P = 0.034).Of the nine patients with pulmonary KS, three of the four in the CG group died of Severe-IRIS-KS, whereas none of the five patients in the EG died . No difference was found in the occurrence of non-severe-IRIS-KS events: 15 in the EG (from 7 patients) and 17 in the CG (from 8 patients) and eight patients in each group did not develop any IRIS event.There were no differences in the number of patients receiving or the number of cycles of vincristine/bleomycin administered. Seven patients in each group did not receive chemotherapy, four in each group had one cycle, eight had two cycles , three patients had three cycles , one patient had four cycles CG and three patients received five cycles .At week-48 follow-up 33 patients were alive and one CG was lost to follow-up. There were no group differences in remission rates; 12 (36.3%) achieved complete remission (six in each group); 15 (45.4.7%) achieved 80–95% remission ; and six (18%) 50–70% remission (three in each group). We did not document any relapses or non-responders.In survivors most measurements improved from baseline to follow-up at week 48; the rate of improvement did not differ between groups. The median CD4+ cells count was 69 cells/mL (IQR 36–119) at baseline and 179 cells/mL (IQR 132–516) at week-48. The median CD8+ at baseline was 715 cells/mL (IQR 395–1178) and 639 cells/mL (IQR 639–673) at week-48. The median CD4+/CD8+ ratio was 0.1 (IQR 0.05–0.22) at baseline and 0.34 (0.16–0.48) at week-48.At baseline, the median of IL-6 was higher for the EG than for the CG: 22 pg/ml; (IQR 15.5–43.9) vs 14 pg/ml, (IQR10.4–23.7); p = 0.02) ; Baseline levels for IL-10 19.7 pg/ml, ; IFN-ϒ 10.4 pg./ml,  were 11.3 pg/ml (IQR 7.4–21), IL10 13.8 (IQR 7.3–36), IFN-ϒ 10.7 pg/ml (IQR 5.6–21.2), and TNF 13.05 pg/ml (IQR 7.7–21.2) there were no differences between groups..1%), both with adherence problems.At week 48, 23 patients (65%) had undetectable HHV-8 VL, 13 in EG, and ten CG. Three had detectable HHV-8 VL in EG (the highest measurement was 929 copies/ml) and six in CG (three of them with HHV-8 VL >10000 copies/ml) p = 0.018) and IL10 76.8 pg/ml, IQR 21–1075 vs. 15.1 pg/ml, IQR 9.9–32, (p = 0.01), CRP 3.4, IQR 3.3–10.1, vs. 1.1, IQR 0.49–1.9, (p = 0.001).The median baseline IL-6, IL-10 and CRP levels in the patients who developed Severe-IRIS-KS was significantly higher than in patients without Severe-IRIS-KS events: IL6 36 pg/ml, IQR 16.1–50 vs. 15.5 pg/ml, IQR 11.7–22 , IFN-ϒ 15.9 pg/ml, IQR 8–82.5 vs. 9.3 pg/ml, IQR 5.8–18.7 (p = 0.1).Complete adverse events are displayed in p = <0.005.The use of Filgastrim was significantly greater in EG OR 3.13 Ten patients acquired a sexually transmitted disease (STD) during the study: eight with syphilis, three at week 24 and five at week 48, one with gonococcal pharyngitis, and one with pediculosis pubis.p = 0.09) in the EG compared to the CG groups, and no reduction on the overall mortality or the number of patients with Severe-IRIS-KS. Exploratory sub-group analysis identified benefits in patients with high baseline HHV-8 viral load, and in those with pulmonary KS, the latter associated with a high risk of death, The sample size was small, due to an overestimation of the impact of valganciclovir, and to unexpected low mortality, probably due to the extensive investigation and treatment of co-infections, not performed before this study. The mortality estimations derived from retrospective studies are likely with underdiagnosis and undertreatment of coinfections and high mortality. KS is an angioproliferative disease caused by HHV-8, mediated by cytokines in severely immunosuppressed individuals [In this randomized clinical trial, we evaluated the effect of valganciclovir on DKS patients who were cART naïve, with severe immunosuppression, comorbid coinfections, and variable levels of HHV-8 viremia. We found a non-significant lower KS attributable mortality , MAC in 10%, parasites, and a diverse combination of HHV-8, CMV, and EBV viremia. In the early years of the AIDS epidemic, similar data were published in DKS case reports; and even it was postulated that these coinfections particularly CMV and other STDs could participate in the pathogenesis of DKS [In this cohort, we found a high prevalence of multiple infections including syphilis 25%), Histoplasmosis and 5%, Histos of DKS . Despites of DKS , only ops of DKS ,47. Coins of DKS .Valganciclovir diminishes HHV-8 VL reducing inflammatory cytokine release when immune reconstitution starts after cART initiation. In Patients with extended disease or Severe-IRIS-KS were treated with 2 mg vincristine and 15 UI bleomycin. This regime was well tolerated; it is not myelosuppressive and does not require concomitant administration of steroids, a recognized amplifier of KS proliferating signaling .The delay in the initiation of cART while giving an anti-HHV8 agent has the same basis as when cART initiation is delayed in CMV chorioretinitis or cryptococcal meningitis, –52 and iOur definition of Severe-IRIS-KS shares similarities to KICS criteria ,54, the vs. none of five in the EG group, a significant difference.DKS patients typically have high mortality, particularly those with pulmonary involvement ,57, in oWe acknowledge some limitations in our study. First, the sample size was small as we overestimated the expected impact of valganciclovir treatment on mortality, and this may explain a mostly negative trial. The studied patients arrived with advanced HIV disease, severely immunosuppressed, an unlikely situation in high-income countries but not in middle and low-income countries where late HIV diagnosis is still an important proportion of newly diagnosed HIV-infected patients. Patients like those described here are likely found in many other middle and low-income countries still nowadays.However, the trial even if considered a pilot study, leads to several interesting new avenues for this group of severely ill patients, including the utility to perform a thorough investigation of co-infections, measuring the viral load of HHV-8 and utilizing an antiviral agent that inhibits its replication.From our perspective is important in patients with DKS to address four elements of KS pathogenesis when initiating treatment: (1) Exhaustive search and treatment of co-infections (2) control of HHV-8 viremia with valganciclovir (or another anti-HHV8 active agent) ,19. In oValganciclovir reduced the number of episodes of Severe-IRIS-KS and also the attributed mortality although this change was not statistically significant.S1 Checklist(DOC)Click here for additional data file.S1 FigThe patients developed three Severe-IRIS-KS events and died.(TIF)Click here for additional data file.S2 Fig(TIF)Click here for additional data file.S3 FigValganciclovir was re-started on week 24; HHV-8 VL replication was finally suppressed. This subject was excluded from the per-protocol analysis.(TIF)Click here for additional data file.S1 File(ZIP)Click here for additional data file.S2 File(PDF)Click here for additional data file.S3 File(PDF)Click here for additional data file.S4 File(DTA)Click here for additional data file.S5 File(PDF)Click here for additional data file.


Dopamine has effects on the mammalian heart. These effects can include an increase in the force of contraction, and an elevation of the beating rate and the constriction of coronary arteries. Depending on the species studied, positive inotropic effects were strong, very modest, or absent, or even negative inotropic effects occurred. We can discern five dopamine receptors. In addition, the signal transduction by dopamine receptors and the regulation of the expression of cardiac dopamine receptors will be of interest to us, because this might be a tempting area of drug development. Dopamine acts in a species-dependent fashion on these cardiac dopamine receptors, but also on cardiac adrenergic receptors. We will discuss the utility of drugs that are currently available as tools to understand cardiac dopamine receptors. The molecule dopamine itself is present in the mammalian heart. Therefore, cardiac dopamine might act as an autocrine or paracrine compound in the mammalian heart. Dopamine itself might cause cardiac diseases. Moreover, the cardiac function of dopamine and the expression of dopamine receptors in the heart can be altered in diseases such as sepsis. Various drugs for cardiac and non-cardiac diseases are currently in the clinic that are, at least in part, agonists or antagonists at dopamine receptors. We define the research needs in order to understand dopamine receptors in the heart better. All in all, an update on the role of dopamine receptors in the human heart appears to be clinically relevant, and is thus presented here. Dopamine is a catecholamine . DopaminHowever, in the peripheral organs, dopamine is also found, and in them, dopamine might be of therapeutic and pathological relevance. In this respect, we will mainly focus our attention into the heart and the circulation. In the context of cardiac function and circulation, the role of dopamine in the kidney is important, and it will also be mentioned in the present review.Dopamine has direct effects on the mammalian heart. These effects can include an increase in the force of contraction, an elevation of the beating rate, and the constriction of the coronary arteries. Depending on the species studied, positive inotropic effects were strong, very modest, or absent, or even negative inotropic effects occurred. We will first start with the formation and degradation of dopamine, we will then move on to dopamine receptors and switch dopamine in cardiovascular pathology, ending with options to use dopamine receptors for cardiac drug therapy. We will always compare animal and human data, stressing on the clinical relevance of the findings. Dopamine  has been called the third endogenous catecholamine, next to adrenaline and noradrenaline e.g., ,17). Dop. Dop17])Convincing data support the presence of dopamine in the mammalian heart. These include the canine left ventricle , in theHow is dopamine then degraded? Dopamine can be converted by a dopamine-β-hydroxylase that is also expressed in the heart  to noradExtracellular dopamine can reach the cytosol of a cell via a dopamine-transporter (DAT) , which i1-, D2-, D3-, D4-, D5-dopamine- receptors, 1-dopamine- and D5-dopamine receptors, which may be regarded as one family (presenting no exons: “D1-like”), have in common that they both stimulate the enzymatic activity of adenylyl cyclases. This holds also true in the heart: In plasma membranes from rat hearts, dopamine and the D1,5-dopamine receptor agonist SKF 38393 , directly (without cAMP) via the βγ-subunits of guanosine triphosphate-binding proteins, some ion channels ) general ,80,144,  general ,177.1-dopamine receptors [2-dopamine receptor, which led to impaired mitochondrial function [1-dopamine receptors in patients with myocardial infarction. Oxidative stress can reduce at least the mRNA for the D1-dopamine receptor [1-dopamine receptor expression [1-dopamine receptors (with Western blotting) in cultivated neonatal rat cardiomyocytes [2-dopamine receptor by raclopride  might be involved in cardiac hypertrophy.Simulated ischemia and reperfusion led in isolated neonatal rat cardiomyocytes to an increased expression (on protein level) of Deceptors , but alsfunction . This mireceptor . The undpression . Hypoxiamyocytes . Blockinclopride B in neoncriptine A, suggesInterestingly, in a genetic model of cardiomyopathy, the cardiomyopathic hamster, when the cardiomyopathy progressed, a decrease in the activity of the β-hydroxylase activity in the heart was reported. Hence, an increase in cardiac dopamine levels seems to correlate in this model with heart failure .2-dopamine receptors using the agonist cabergoline , the blood–brain barrier may open, which is detrimental. The stimulation of the Dergoline A, 20 µM,eceptors . Hence, 1-dopamine receptors (and D2-dopamine receptors) was enhanced on a protein level. Fittingly, the phosphorylation state of DARPP32, the phosphorylation state of the cAMP-dependent protein kinase  in rats, dopamine turned out to be less potent and effective than the pure β-adrenoceptor agonist dobutamine, compared with rats without sepsis . Here, t1-dopamine receptors in the brain  can be blocked by many antipsychotic drugs  or propylbutyldopamine (a D2-dopamine receptor agonist), suggesting that the D1-dopamine receptor and the D2-dopamine receptor in the human heart can directly or indirectly increase the force of contraction [Lower concentrations of dopamine could theoretically dilate the coronary vessels via Doceptors . Moreoveoceptors . Higher h sepsis . There atraction .1-dopamine receptors in patients with heart failure, compared to the matched controls [1-dopamine receptor simulation, compared to β-adrenergic stimulation. Data in this regard are also not yet convincing.A Japanese group convincingly reported on an increased cardiac expression of the mRNA for the Dcontrols . Howevercontrols . Moreove2-dopamine receptors, also have agonistic effects on D1-dopamine receptors [2-dopamine receptor agonists such as bromocriptine or pergolide have a higher potency on D2-dopamine receptors than D1-dopamine receptors. However, pergolide and bromocriptine have an affinity of 0.1 µM (as Ki) or 0.6 µM at D1-dopamine receptors; therefore, they can act as agonists at the D1-dopamine receptors [All dopamine agonists clinically used to treat Morbus Parkinson, besides the intended agonistic effects on Deceptors ,185 . Genetic models in mice have been helpful in this regard, and new genetic models might in many ways add to our knowledge of the cardiac function and the underlying signal transduction mechanisms in the human heart. These mice models might be used in subsequent work to obtain models for cardiovascular diseases such as arrhythmia, genetically induced heart failure, septic heart failure, and drug-induced heart failure. Studying these mechanism, it should be possible to repurpose known agonists and antagonists at the D1,5-dopamine receptors, or to invent new drugs that are biased agonists at relevant dopamine receptors, or to inhibit deleterious signal transduction pathways. More effort might be put into the study of “D2-like” receptors in the heart, where only very scarce data seem to be available.There is growing evidence that dopamine cannot only act on the contractile function of the heart, especially the human heart, by directly or indirectly stimulating α- and β-adrenoceptors. In addition, dopamine seems to stimulate the D1-dopamine receptors, is a timely and promising endeavor in experimental and clinical cardiovascular pharmacology. We hope that these efforts will improve patient care, and possibly reduce cardiovascular morbidity and mortality in the years to come.In summary, this review tried to convince the reader that the study of dopamine in the heart, and a fresh look at cardiac dopamine receptors, especially D


One of the hallmarks of Parkinson’s disease (PD) is the progressive loss of dopaminergic neurons and associated dopamine depletion. Several mechanisms, previously considered in isolation, have been proposed to contribute to the pathophysiology of dopaminergic degeneration: dopamine oxidation-mediated neurotoxicity, high dopamine transporter (DAT) expression density per neuron, and autophagy-lysosome pathway (ALP) dysfunction. However, the interrelationships among these mechanisms remained unclear. Our recent research bridges this gap, recognizing autophagy as a novel dopamine homeostasis regulator, unifying these concepts. I propose that autophagy modulates dopamine reuptake by selectively degrading DAT. In PD, ALP dysfunction could increase DAT density per neuron, and enhance dopamine reuptake, oxidation, and neurotoxicity, potentially contributing to the progressive loss of dopaminergic neurons. This integrated understanding may provide a more comprehensive view of aspects of PD pathophysiology and opens new avenues for therapeutic interventions. Parkinson’s disease (PD) is a progressive neurodegenerative movement disorder. While mainly characterized by motor syndrome, PD is associated with non-motor symptoms (NMS) in almost all patients. More than 10 million primarily older adults worldwide suffer from PD. The most common symptom associated with PD is tremor, which typically begins on one side before eventually affecting both sides over time. Other symptoms may include muscle rigidity (stiffness), gait disturbances , slowness/decreased mobility, impaired speech/swallowing problems, depression/anxiety disorders, hyposmia, constipation, urinary dysfunction, orthostatic hypotension, pain, sleep disturbances, and cognitive decline, including dementia or psychosis later in life if left untreated for too long. While there is no cure for PD, symptomatic dopaminergic therapy helps improve the quality of life for those with the disease. PD is characterized by the presence of certain hallmarks in its pathology. These include progressive selective loss of dopaminergic neurons in the midbrain substantia nigra pars compacta (SNpc); Lewy bodies containing aggregates of α-synuclein protein; gliosis or inflammation around affected areas; and a dramatic decrease in dopamine, a neurotransmitter associated with movement and reward. While the motor syndrome of PD is related to nigral degeneration and dopamine depletion, NMS may be linked to the degeneration of other neural types, including the peripheral autonomic nervous system . PD is aThe molecular pathogenesis of PD involves a complex interplay of several factors, such as impaired mitochondria, protein homeostasis, autophagy-lysosome pathway (ALP) dysfunction at the synapse, trafficking of the dopamine transporter (DAT), DA toxicity, oxidative stress, disruptions in synaptic vesicle endocytosis, autonomous pacemaking, calcium homeostasis imbalance, iron-content, extensive axonal arborization size, prion-like α-synuclein transmission, and neuroinflammation. The reader is referred to several review articles that discuss these pathophysiological mechanisms in detail . AcknowlThe ALP plays a crucial role in the pathogenesis of PD. Autophagy, or self-eating, is a process by which cells degrade and recycle their cellular components, such as organelles, proteins, lipids, and nucleic acids, for use in metabolic processes. The autophagic process is regulated by several molecular pathways involving the lysosomes and other organelles, including the endoplasmic reticulum and mitochondria . The lysMacroautophagy: characterized by the formation of a double membrane organelle, the autophagosome, which forms around its cargo and transports it to the lysosome. There are two types of macroautophagy in terms of cargo recognition: selective and non-selective. Relevant examples of selective macroautophagy include mitophagy (mitochondria) and aggrephagy (protein aggregates), among many others .Microautophagy: characterized by the lysosome’s direct internalization of small cytoplasmic cargo. Microautophagy can also be selective and non-selective .Chaperone-mediated autophagy (CMA): characterized by selective recognition of cytoplasmic proteins harboring a KFERQ or KFERQ-like motif by chaperones that escort them to the lysosome where they undergo translocation through the lysosomal membrane to its lumen for degradation .In PD, the ALP pathway has been found to be dysregulated in both familial  and sporSeveral genes related to ALP have been implicated in PD due to their mutations. These genes have roles in various aspects of the pathway, from regulating autophagy initiation to the proper functioning of lysosomes. Here are a few examples of some key genes:LRRK2) mutations are one of the most common genetic causes of autosomal dominant PD. The LRRK2 protein is involved in various cellular processes, including autophagy and endolysosomal functions. Mutations can lead to impaired autophagy and increased susceptibility to PD  is a retromer complex component. VPS35 mutations have been linked to autosomal dominant PD. VPS35/retromer complex is essential for the retrieving specific membrane proteins from endosomes to the trans-Golgi network, thereby maintaining lysosomal function .ATPase 13A2 (ATP13A2), also known as PARK9, is a lysosomal P-type ATPase. Mutations in this gene are associated with a rare form of early-onset parkinsonism (Kufor-Rakeb syndrome). ATP13A2 is involved in maintaining lysosomal pH and manganese homeostasis, and its dysfunction contributes to impaired autophagy .PTEN-induced putative kinase 1 (PINK1) mutations are associated with autosomal recessive early-onset PD. PINK1 is a mitochondrial kinase that plays a key role in mitophagy, a selective form of autophagy responsible for removing damaged mitochondria .Parkin RBR E3 Ubiquitin Protein Ligase (PARKIN) mutations are also linked to autosomal recessive early-onset PD. PARKIN functions as an E3 ubiquitin ligase and works with PINK1 in the mitophagy pathway, promoting the clearance of damaged mitochondria .Protein deglycase DJ-1, also known as Parkinson’s disease protein 7 (DJ-1), mutations are implicated in autosomal recessive early-onset PD. DJ-1 has multiple roles in the cell, including acting as a redox-sensitive chaperone and a sensor for oxidative stress. DJ-1 is also involved in regulating autophagy and mitophagy .Auxilin (DNAJC6/PARK19) is a cofactor for uncoating clathrin-coated vesicles for the heat shock cognate protein-70 (Hsc70) . Its losSYT11  dopaminergic neurons are moderately affected. The dopaminergic neurons in SNpc are most affected in PD. In contrast, the dopaminergic neurons spared in PD patients’ postmortem tissues show lower DAT expression per neuron than controls. For example, Uhl et al. reported that DAT mRNA per neuron is higher in SNpc compared to the adjacent nucleus paranigralis of the ventral tegmental area (VTA) in neurologically normal study subjects. Surviving SNpc neurons in age-matched PD patients is almost half that in controls . The samty to PD .in vivo.Consistent with the observations mentioned above, preclinical evidence demonstrates that ectopic or elevated DAT expression is sufficient to induce neurodegeneration. Chen et al. generated a Tet-inducible forebrain DAT-transgenic mouse model under the calcium/calmodulin-dependent protein kinase type II alpha (CaMKIIα) promoter. CamDAT mice spontaneously develop motor dysfunctions and progressive neurodegeneration in the striatum, which l-DOPA accelerates . Masoud One of the hallmarks of PD progression is the reduction of DAT levels in the brain, which is the direct result of the loss of DA terminals. However, several studies observed increased DAT levels without or preceding dopaminergic neurodegeneration. Here I list examples of these studies.LRRK2 and GBA mutations enrolled into the Parkinson’s Progression Markers Initiative (PPMI) focused on 123-I Ioflupane DAT imaging reported that non-manifesting carriers of GBA mutation had increased DAT binding in all striatal regions compared with healthy controls and non-manifesting carriers of the LRRK2 mutation (GBA gene) regulates autophagy  as assessed by 123I-PE2I single-photon emission computerized tomography scans and confirmed by post-mortem immunohistochemical analyses. This non-human primate model induces Parkinson’s disease-like pathology, including α-synuclein aggregation with pathological properties such as amyloid-like Lewy body pathology in PD brains and nigral neuronal degeneration . Similarn levels . Furthern levels .LRRK2 G2019S mutant knock-in (GKI) mice show a significant increase in striatal DAT protein as assessed by western blot  and dopamax) in the dorsal but not ventral caudate-putamen  quickly loads dopamine into synaptic vesicles after its synthesis in the cytoplasm or reuptake in the presynaptic terminals by DAT. In addition to dopamine sequestration, the acidic pH ~5.6 of synaptic vesicles stabilizes dopamine . DopaminSeveral dopamine–quinone molecules can undergo oxidative ligation to α-synuclein, selectively inhibiting the protofibril-to-fibril conversion . α-synucCatecholaldehyde hypothesis,” depicting the role of DOPAL in SNpc dopaminergic degeneration in PD (Monoamine oxidase (MAO) enzymatically metabolizes cytoplasmic dopamine to 3,4-dihydroxyphenylacetaldehyde (DOPAL), a toxic metabolite . Under pon in PD .in vitro and in vivo. The rapid induction of autophagy by cocaine is evident in tyrosine hydroxylase-positive nerve terminals located in the nucleus accumbens (NAc), a brain region implicated in reward, motivation, and addiction. Remarkably, this induction of autophagy can be detected as early as 3 min after systemic cocaine injection. Cocaine administration leads to DAT depletion in dopaminergic nerve terminals in the NAc. The depletion of DAT by cocaine in the nucleus accumbens nerve terminals has been detected using various experimental techniques. Western blot analysis of isolated nerve terminals demonstrates a reduction in DAT protein levels, while immunostaining of brain sections reveals decreased DAT expression in dopaminergic neurons. Furthermore, dopamine uptake activity in isolated nerve terminals, assessed using radiolabeled dopamine (3H-dopamine), shows a reduction in the maximum velocity of dopamine uptake, confirming the depletion of DAT from the nerve terminals’ membranes. Interestingly, pharmacological inhibition of autophagy reverses the depletion of DAT induced by cocaine. This suggests that cocaine-mediated autophagy plays a significant role in modulating DAT levels and dopaminergic neurotransmission (Recently, our studies have suggested that DAT can be selectively targeted for autophagic degradation . Our finsmission .Selective degradation of DAT by autophagy represents a novel mechanism for regulating dopamine signaling and dopaminergic neurotransmission. By modulating DAT levels, autophagy can control dopamine reuptake, its availability in the synaptic cleft, and its potential accumulation in the dopaminergic neurons’ cytoplasm in case of sustained elevated dopamine reuptake. This has important implications for the strength and duration of dopaminergic signaling and dopamine neurotoxicity.The above-mentioned abnormal mechanisms have been proposed to contribute to PD pathophysiology. These include:Dopamine oxidation-mediated neurotoxicity. The catechol structure of dopamine makes it particularly susceptible to enzymatic and non-enzymatic oxidation, generating dopamine quinones and other ROS. These oxidative species can damage cellular components, disrupt mitochondrial function, and initiate cell death pathways, contributing to the neurodegeneration seen in PD.High expression density of DAT per neuron in SNpc. The extent of dopaminergic neuron loss in PD has been found to correlate strongly with the expression density of DAT per neuron. High DAT density could increase dopamine reuptake and intracellular accumulation, promoting dopamine oxidation and associated neurotoxicity. However, the precise role and regulation of DAT in PD pathogenesis have remained unclear.Dysfunction of the autophagy-lysosome pathway. The ALP has also been implicated in PD. Dysregulation of this pathway can lead to the accumulation of damaged proteins and organelles, including potentially neurotoxic dopamine metabolites. Although the connection between autophagy dysfunction and PD has been established, its implications for dopamine homeostasis and DAT regulation were not fully understood.However, until recently, these mechanisms have been studied in isolation, with no apparent connection to link them together.Our recent research has provided a unifying framework that integrates these previously separate mechanisms. We have discovered that autophagy is a novel mechanism for controlling dopamine homeostasis, directly linking autophagy-lysosome pathway dysfunction, DAT density, and dopamine oxidation. Autophagy declines with aging , an obseIn conclusion, our research bridges the gap between three previously separate concepts in PD pathophysiology, providing a more comprehensive understanding of aspects of the disease’s pathogenesis. By recognizing autophagy as a novel regulator of dopamine homeostasis, a missing link is recognized between dysfunctional ALP, high expression density of DAT per neuron, dopamine oxidation-mediated neurotoxicity, and correlation with vulnerability of dopaminergic neurons in PD.The author confirms being the sole contributor of this work and has approved it for publication.The author declares that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


As a result, data mining has become a widely used technique in infodemiology and infoveillance research to combat misinformation effects. On the other hand, there is a lack of studies that specifically aim to investigate misinformation about fluoride on Twitter. Web-based individual concerns on the side effects of fluoridated oral care products and tap water stimulate the emergence and propagation of convictions that boost antifluoridation activism. In this sense, a previous content analysis–driven study demonstrated that the term This study aimed to analyze “fluoride-free” tweets regarding their topics and frequency of publication over time.A total of 21,169 tweets published in English between May 2016 and May 2022 that included the keyword “fluoride-free” were retrieved by the Twitter application programming interface. Latent Dirichlet allocation (LDA) topic modeling was applied to identify the salient terms and topics. The similarity between topics was calculated through an intertopic distance map. Moreover, an investigator manually assessed a sample of tweets depicting each of the most representative word groups that determined specific issues. Lastly, additional data visualization was performed regarding the total count of each topic of fluoride-free record and its relevance over time, using Elastic Stack software.We identified 3 issues by applying the LDA topic modeling: “healthy lifestyle” (topic 1), “consumption of natural/organic oral care products” (topic 2), and “recommendations for using fluoride-free products/measures” (topic 3). Topic 1 was related to users’ concerns about leading a healthier lifestyle and the potential impacts of fluoride consumption, including its hypothetical toxicity. Complementarily, topic 2 was associated with users’ personal interests and perceptions of consuming natural and organic fluoride-free oral care products, whereas topic 3 was linked to users’ recommendations for using fluoride-free products  and measures , comprising the propaganda of dental products. Additionally, the count of tweets on fluoride-free content decreased between 2016 and 2019 but increased again from 2020 onward.Public concerns toward a healthy lifestyle, including the adoption of natural and organic cosmetics, seem to be the main motivation of the recent increase of “fluoride-free” tweets, which can be boosted by the propagation of fluoride falsehoods on the web. Therefore, public health authorities, health professionals, and legislators should be aware of the spread of fluoride-free content on social media to create and implement strategies against their potential health damage for the population. Social media have increasingly gained popularity as platforms for individuals and communities to seek and share health information. In that regard, Twitter boasts an impressive user base of approximately 368 million monthly active individuals who share a common interest in discussing common health topics, such as COVID-19, cancer, influenza, mental health, vaccination, and smoking . AlthougAlthough Twitter can contribute to spreading health falsehoods, it can also serve as a valuable tool for elucidating the social factors that contribute to the onset of negative beliefs. As a result, data mining has become a widely used technique in infodemiology and infoveillance research to combat misinformation effects ,13,14. Ffluoride-free was frequently associated with antifluoridation interests on social media [Since its introduction in 1942, multiple studies have consistently demonstrated the role of fluoride in reducing the prevalence and incidence of caries lesions . There ial media .In view of possible negative implications of web-based fluoride content, this study aimed to analyze “fluoride-free” tweets regarding their topics and frequency of publication over time.This study did not require institutional review board approval from the Council of Ethics in Human Research of the University of Waterloo because federal regulations do not apply to research using publicly available data that do not involve human subjects.On June 5, 2022, the Twitter application programming interface  was usedThe CSV file data set was uploaded and processed to enhance the quality of the tweets for subsequent analysis . InitialFurthermore, exploratory data preprocessing was performed to assess the impact of both the most frequent and rare words in the data set. Notably, term frequency filtering did not exhibit substantial improvements on latent Dirichlet allocation (LDA) topic modeling. Therefore, the decision was made to retain the most frequent and rare words to enhance the contextual relevance of the resulting topics .LDA topic modeling is an unsupervised machine learning probabilistic and word count–based algorithm used to effectively determine salient terms and extract emerging groups (topics) of a particular corpus (data set), including social media posts concerning health issues ,32. EvenGensim and spaCy libraries in a Google Colaboratory interface to identify the salient terms and topics [The preprocessed 21,169 tweets were subjected to LDA topic modeling techniques using the Python-based d topics ,32. The d topics .K topics, where K ranged from 2 to 50, before eventually narrowing the consideration range down to 3 to 15 cases. Remarkably, higher coherence scores probabilistically represent better-quality topic modeling, facilitating the interpretation of outputs. An investigator (ML) validated the topic model with the highest coherence score by rating the model on the meaningfulness of its topics, how well publications within a single topic are related, and how publications of different topics are distinct.In this context, the coherence values were computed for x,y) coordinates.Facing the definition of the number of topics, the similarity between them was calculated through an intertopic distance map. It is a visualization of the issues in a bidimensional space, where the bubbles represent topics that emerged from the topic modeling analysis and are dimensionally proportional to the number of contained words. In this graph representation, the axes are based on 2 principal components to facilitate the interpretation of the distance between topics. Synthetically, the LDA algorithm uses a principal component analysis to reduce N-dimensional vectors to bidimensional vectors and achieve  manually assessed a sample of tweets depicting each of the most representative word groups that determined specific issues. Lastly, additional data visualization was performed regarding the total count of each topic of fluoride-free record and its relevance over time, using Elastic Stack software through the Elasticsearch and Kibana features -38.To determine the underlying social issues surrounding the spread of fluoride-free content on Twitter, we applied topic modeling to identify patterns based on the frequency of keywords ,30. A hiUsing an intertopic distance map , we visuTo determine the frequency that each “fluoride-free” topic appears on Twitter, we calculated the percentage  and absoThe relevance of each topic was determined by the number of tweets over time. The points of each curve are separated by a 4-week interval . The numThe purpose of this study was to analyze the fluoride-free content on Twitter. Although studies have investigated fluoride-related content on Facebook, Instagram, and Twitter ,28,39-42Despite a decline in “fluoride-free” tweets between 2016 and 2019, the antifluoridation movement has had a negative impact on dental public policies in English-speaking countries. This has led to some regions discontinuing community water fluoridation to meet specific demands of users, despite community-based intervention being a key factor in promoting oral health equity . For exaFalse health beliefs usually originate from the same mechanisms of accurate health beliefs, influenced by diverse cognitive, social, and affective drivers such as intuitive thinking , cognitive failures (neglecting or forgetting sources and evidence), illusory truth , reliance on source cues , emotion , and worldview  ,50. NotaThe findings may contribute to elaborating educational public health actions regarding eHealth and mobile health solutions, aiding communities in avoiding antifluoridation content on social media. Likewise, these results can orient fact-checking agencies to promptly clarify deceitful web-based publications about fluoride to their audience, addressing the concerns of users to support fluoride-containing measures. For example, issues concerning fluoride overexposure must be clarified, reinforcing safeguard actions for storing and using fluoridated products in early childhood ,25. CertThis study has some limitations. First, we used only data from Twitter for the proposed analyses. As a result, the findings are influenced by platform characteristics , differentiating them from other social media. Despite that limitation, previous studies also focused on Twitter data to investigate health topics on social media because of the advantages relating to data collection and the number of active users ,59. SecoThe public concerns toward a healthy lifestyle, including the adoption of natural and organic oral care products, seem to be the main motivation of the recent increase of “fluoride-free” tweets, which can be boosted by the propagation of fluoride falsehoods noted on the web. Therefore, public health authorities, health professionals, and legislators should be aware of the spread of fluoride-free content on social media to create and implement strategies against their potential health damage for the population.


Dural defects are a common problem in neurosurgical procedures and should be repaired to avoid complications such as cerebrospinal fluid leakage, brain swelling, epilepsy, intracranial infection, and so on. Various types of dural substitutes have been prepared and used for the treatment of dural defects. In recent years, electrospun nanofibers have been applied for various biomedical applications, including dural regeneration, due to their interesting properties such as a large surface area to volume ratio, porosity, superior mechanical properties, ease of surface modification, and, most importantly, similarity with the extracellular matrix (ECM). Despite continuous efforts, the development of suitable dura mater substrates has had limited success. This review summarizes the investigation and development of electrospun nanofibers with particular emphasis on dura mater regeneration. The objective of this mini-review article is to give readers a quick overview of the recent advances in electrospinning for dura mater repair. Electrospinning technology, as an excellent method for producing nano-to-micro-scale fibers, has attracted much attention due to its versatility, rapidness, controllability, low cost, and large surface area to volume ratio micro/nanofibers with uniform or special microscopic morphology ,2,3. EleA general electrospinning setup consists of four primary components: a high-voltage power supply, a capillary tube, a spinneret, and a grounded collector  13]. In. In13]. (i)Electrospun nanofibers have a diameter ranging from nano to micro scale.(ii)Electrospun nanofibers possess high porosity.(iii)The fibers produced by the electrospinning technique have a large aspect ratio and a high surface-to-volume ratio.(iv)Electrospun nanofibers possess superior mechanical properties and flexibility.(v)The electrospinning process enables the production of nano/microfibers with an infinite number of chemical compositions.(vi)Various types of morphology can be prepared by modifying the spinneret.With the increasing popularity of nanofibers, electrospinning technology has been employed extensively. So far, more than 200 polymers have been electrospun into the nanofiber form ,14. The Due to the aforementioned characteristics of electrospun nanofibers, they can be applied for numerous applications such as catalysis, energy storage, environmental remediation, filter media, sensor, textile, protective clothing, biomedicals, fire retardant, etc. ,27,28,29In order to widen the applications of electrospun nanofibers in the biomedical field, a new form called ‘cell electrospinning’ (C-ES) was developed by Townesend-Nicholson et al. in 2006 . The aut-It must be biocompatible.-It should mimic the physiochemical structure of the dura mater.-It should adequately restore the continuity of the dura mater and prevent the leakage of CSF.-It should lower the chances of infection.-The mechanical properties of the substrate should facilitate the suturing.-It should enhance tissue regeneration.-It should promote the adhesion and migration of dural fibroblasts.-It should minimize local tissue inflammation.-It should not induce adverse reactions.-It should be safe, inexpensive, and easy to handle.The brain is surrounded by three protective meningeal layers . The dura mater is the outermost layer that surrounds and protects the brain and spinal cord . It is m-Biocompatibility without inducing an immune or inflammatory response.-Controlled risks of infection.-Appropriate mechanical properties, which are resistant to tearing-Anti-leakage of CSF.The US Food and Drug Administration (FDA)  and The National Medical Products Administration of China  suggest that the ideal dural substitute should include the following attributes :-BiocompIn the last decade, new artificial polymeric dural substitutes have been developed, some of which are in clinical trials. Nanoabsorbable synthetic polymers such as silicon and expanded polytetrafluoroethylene have been used for the dural implant; however, it may cause serious complications such as induction of granulation tissue formation due to long-term foreign body reaction ,70. Natu4HB) possessing good mechanical strength, flexibility, wettability, and biocompatibility via chemosynthesis and electrospinning methods [It is of great significance to develop an artificial dura substitute that matches the intrinsic structure and mechanical properties of natural dura mater. Zwirner et al.  investig methods . The in Control of the fiber orientation is highly desirable to obtain increased complexity and performance . RecentlIntegration of two or more materials in a composite hold great potential in dural repair . The comDeng et al.  designedn-octyl-2-cyanoacrylate (NOCA), a medical glue, into fiber membrane for treating dural defects. The in vitro and in vivo experiments were carried out on egg membranes and goat meninges, respectively. The results showed that the NOCA membranes had properties of high strength, good flexibility, and waterproofness without leakage  and investigated for dural implants.Although electrospun nanofiber membranes can serve as an effective scaffold for dura repair, there are a few limitations that hinder their application in this field. For example, the dura substitute should mimic the intrinsic structure of the human dura mater. It is challenging to fabricate an electrospun nanofiber membrane with precise dimensions and morphology. Poor mechanical properties and rapid degradation, particularly in the case of natural polymers, are also a challenge to overcome. Authors have attempted to address this issue by fabricating composite nanofibers from natural and synthetic polymers. However, it is difficult to obtain properly blended natural-synthetic composite nanofibers due to the poor miscibility of the component polymers. In some cases, the solvents may alarm additional issues such as miscibility, biocompatibility, pungent smell, etc. Aligned nanofiber membranes have been developed to enhance mechanical properties and cell growth. However, it may need a modified electrospinning setup. Another concern is the adhesion formation between the scaffold membrane and overlying tissue. In order to obtain proper and fast growth of nerve cells, neurotropic factors-incorporated nanofibers membranes have been developed. The potential side effects and achieving controlled release are the major concerns. Therefore, several technical and pharmacological issues should be addressed prior to exploiting their potential. Safety is another issue in electrospinning as high voltage is used during its operation.It is evident from the literature that most of the studies are at the lab scales, and further studies are needed for their clinical applications. It means that there is still a need for an optimized dural substitute with safety, proper mechanical properties, and low cost. By optimizing various properties of nanofibers membranes in terms of fiber orientation, morphology, porosity, drug release, mechanical properties, and, most importantly, biocompatibility, the product can fulfill specific requirements. With the rapid development of materials science and ongoing research on the dura substitutes, it can be hoped that new implant materials will be developed soon, which can overcome all the limitations of the currently available products.


Antimicrobial Resistance (AMR) is one of the top ten global public health threats facing humanity, alongside climate change. Here, we aim to summarise the effects of climate change  on spread of antibiotic resistance and on infections with antibiotic-resistant bacteria in Germany.We conducted a literature search with articles published between January 2012 and July 2022. Two authors screened titles, abstracts and full texts and extracted the data systematically.From originally 2,389 titles, we identified six studies, which met our inclusion criteria. These studies show that an increase in temperature may lead to higher antibiotic resistance rates and an increased risk of colonisation as well as spread of pathogens. Furthermore, the number of healthcare-associated infections increases with increased temperature. Data indicate that higher antibiotic use is present in areas with warmer mean temperature.European data are scarce, but all studies identified point towards an increasing AMR burden due to climate change. However, further studies are needed to draw attention to the links between climatic factors and AMR and develop targeted preventive measures. Climate change has a clear impact on disease burden globally, and there is sufficient evidence to act now to mitigate the climate crisis. However, it remains challenging to quantify the impact of climate change on health for many diseases. The complexity of climate-dependent modifying factors of infectious diseases challenges the precision of modelling pathways and subsequent impacts.Escherichia coli (E. coli), meticillin-resistant Staphylococcus aureus (S. aureus), carbapenem-resistant Pseudomonas aeruginosa (P. aeruginosa) and third-generation cephalosporin-resistant Klebsiella pneumoniae (K. pneumoniae). These findings were recently confirmed by Mestrovic et al. [Since the availability of antimicrobials, infections due to bacteria, viruses, fungi and parasites have become treatable. However, due to the misuse and overuse of antimicrobials, these pathogens become increasingly resistant. In 2019, the World Health Organization (WHO) declared antimicrobial resistance (AMR) one of the top ten global public health threats facing humanity, alongside climate change . In Euroc et al.  in anothc et al. . Furtherc et al. .S. aureus, E. coli and K. pneumoniae increased with increasing temperature.Temperature, which may increase due to climate change , is knowIn this review, we aim to summarise the key effects that climate change may have on the spread and burden of AMR among humans in Germany and Europe. We conducted a literature review and adapted a graphical model to support this assessment.We conducted a literature search limited to peer reviewed articles published between January 2012 and July 2022 in English, French or German. PubMed was searched using the keywords climate change, temperature, healthcare-associated infection (HAI), and antimicrobial resistance, including the MeSH terms for climate change, global warming, climatic processes, antibacterial agents, and drug resistance . The complete search string is documented in the Two authors (AM and FW) independently of each other screened titles, abstracts and full texts and extracted the following data systematically: citation, study period, study design, demographics, climate indicators, definition of outcome as pathogen and/or antimicrobial resistance and/or infections analysed. Discrepancies between the reviewers were solved by discussion until a consensus was reached. Data were summarised in tables and stratified by drug resistance, pathogen and type of infection.Additionally, we utilised the webtool provided by Mora et al.  to furthOur search identified a total of 2,389 titles . After eE. coli and K. pneumoniae increased from 2000 to 2016, whereas meticillin-resistant S. aureus (MRSA) decreased over time. The reduction of MRSA detection is commonly attributed to targeted public action and, thus, may not be explained by climate indicators [E. coli (MREC) and carbapenem-resistant K. pneumoniae (CRKP). With increases in mean temperature, the rates of MRSA, MREC and CRKP were shown to increase [E. coli and 0.55% per year for third-generation cephalosporin-resistant E. coli was observed. For fluoroquinolone-resistant E. coli, an increase of 0.57% per year was found after accounting for other recognised resistance drivers including antibiotic consumption and population density. For K. pneumoniae, even higher increases were identified: 0.9% per year for third-generation cephalosporin-resistant K. pneumoniae and 1.2% for fluoroquinolone-resistant K. pneumoniae. McGough et al. [Using data from the European Antimicrobial Resistance Surveillance Network (EARS-Net), McGough et al.  showed tdicators . Kaba etdicators  and McGodicators  show a cincrease . Furtherincrease  demonstrh et al.  concludeh et al.  also conClostridioides difficile, varying by the Southern and Northern hemisphere. Although they found no data supporting associations of temperature, humidity or seasonality with carbapenem-resistant Enterobacteriaceae (CRE), they hypothesised that such associations could also be plausible [The optimal growth temperature for many relevant bacteria is above 30°C , so incrlausible . Surveillausible , 18 and lausible  suggesteS. aureus, Enterococcus spp. and E. coli [Acinetobacter baumannii per 1°C increase and a 4% risk for SSI with Enterobacter spp. in Germany. There was no association found between risk of SSI with Streptococcus spp. or Candida albicans and an increase in temperature.There is also evidence that the number of HAIs increases with increased temperature. This is of relevance because 75% of the AMR burden stem from HAIs, which result in higher use of antibiotics . Surgica E. coli . In part E. coli  showed t E. coli . In a se E. coli  found anStreptococcus pneumoniae with a reduced incidence rate ratio (IRR) of 0.498 (95% Cl: 0.174–1.429) for temperatures ≥20°C compared to temperatures <5°C. This may well be explained by its transmission pathway via droplets, which is more relevant when the population stays predominantly indoors during colder seasons. In addition to temperature, relative humidity inversely correlated with an increase in ha-BSIs [Another study by Schwab et al.  reportedIn the European surveillance study, McGough et al.  presenteE. coli (with 25 publications), followed by S. aureus , P. aeruginosa (five publications) and K. pneumoniae (four publications). For these pathogens, the associated climatic hazards were floods, storms, warming, precipitation, and droughts. The main routes of transmission were waterborne transmission as well as unspecified transmission.In a recent study on the impact of climate hazards on pathogenic human diseases, Mora et al.  collecteClimate change, through increases in temperature and changes in humidity and precipitation, will likely lead to bacterial pathogen spread, increased use of antibiotics and increased AMR in Europe. However, due to the complexity of interactions and the significance of various factors, we were not able not elucidate the full dimension of these associations. European data are scarce, and we were only able to identify six studies that met our inclusion criteria. However, all six of these studies pointed towards an overall further acceleration of increasing AMR burden due to climate change.S. aureus were more frequent in summer and autumn compared to colder seasons. Streptococcus spp. infections were more common in spring and summer months. Another surveillance paper concentrating on AMR genes and their distribution on a global level, taking climatic factors into account, found an uneven distribution of AMR genes in bacteria across several cities worldwide, suggesting an association with higher temperatures in different regions [There is some evidence specifically demonstrating the increase in AMR with increasing temperatures in Europe , 14. Stu regions . More evCandida auris, a fungal pathogen that is often multi-drug-resistant and exists in the environment, may become increasingly pathogenic due to climate change [Climate hazards can bring humans and disease-causing organisms closer together, leading to a rise in pathogen transmission and infections, as demonstrated in our illustrated model with data from Mora et al.  Figure . An incre change . In addie change , 29.Human pathogen colonisation and infection can result from exposure to pathogens across a range of community and healthcare system domains and may be influenced by socioeconomic and environmental determinants of health. With our focus on human health, we did not comprehensively assess the changes in AMR from a One Health perspective, although we partly highlight this aspect by taking the review by Forrester et al.  into accPossible limitations of our analysis are related to the limited number of studies included in this review. Among the excluded 35 non-European studies, there may be additional relevant evidence to better understand the association of climate change and AMR.For included studies, the predominant study type was observational, where surveillance data of pathogens or infection were combined with climate data in form of an ecological study design. Here, measuring errors may occur in exposure and outcome leading to risk of bias and imprecision of the reported estimates. Even though various confounders were addressed within the single studies, there may be additional factors with influence on the association of climate and AMR or HAIs. By focusing on EU/EEA countries in this review, confounding due to societal and economic factors may have been reduced. Nonetheless, their potential relevance for AMR has been demonstrated before and should be kept in mind.The two publications by Aghdassi et al. , 18 inclThe links between AMR and climate change require significantly more attention. Further detailed studies with local data on weather conditions, resistance situation, infectious diseases, but also antibiotic use, social determinants and other infectious and social factors are needed to better understand the pathways and subsequent impacts of these associations. Financial and scientific resources need to be made available to realise these studies. The example of decreasing MRSA incidence in many EU countries due to targeted public health action, in particular infection prevention and control measures in hospitals, is a reassuring sign that the AMR pandemic can be controlled. But climate change will likely make the control of AMR more challenging. On the other hand, there may be synergistic effects (or co-benefits) when leveraging action against both AMR and climate change, for example through reduction of industrial livestock farming or meat production and consumption. Although this summary focused on evidence in Europe, AMR is a pandemic threat. The Global South often suffers from a higher burden of AMR. Thus, prevention and control of AMR will have limited success with single country interventions and needs a global focus.Despite the complexity of climate change processes and AMR, it will be important to closely monitor the changes in AMR over time in order to inform the prioritisation of public health measures. Intensified One Health surveillance approaches will be needed. Global surveillance systems such as the European EARS-Net and the worldwide Global Antimicrobial Resistance and Use Surveillance System (GLASS) are particularly important for country comparisons and could enable ecological studies analysing associations with regional climate change. Globally, we will have to further improve the availability and standardisation of antimicrobial susceptibility testing (AST), particularly in the Global South, to allow for more representative AMR surveillance data. In addition, AMR burden of disease studies will have to be developed and routinely updated to inform timely public health decision-making.In addition to improved One Health AMR surveillance, universal health coverage and effective infection prevention and control measures including reliable access to water, sanitation and hygiene and One Health antimicrobial stewardship are needed to control the AMR pandemic worldwide. Investment in research and development on new antimicrobial drugs is needed as well as on the development of relevant vaccines.Climate change and AMR are two of the biggest and most complex threats facing the world. Both have been exacerbated by, but can be contained through, human action.Increases in temperature and changes in humidity and precipitation may increase the spread of AMR and healthcare-associated infections.Climate change contributing to more frequent and severe extreme weather events will likely result in increased antimicrobial drug use in humans and animals.Overuse of antimicrobial drugs across human, animal and environmental sectors exacerbates AMR.More financing, political advocacy and coordinated global action are needed to better respond to the threats of antimicrobial resistance and the climate crisis.


Rasagiline added in 10−13 to 10−5 mol/L concentrations did not alter the resting or electrically stimulated [3H]dopamine release. Rasagiline (10−9 mol/L), however, suspended the stimulatory effect of selegiline on the electrically induced [3H]dopamine release. The trace amine-associated receptor 1 (TAAR1) antagonist EPPTB (10−8–10−7 mol/L) also inhibited the stimulatory effect of selegiline on [3H]dopamine release. The effect of selegiline in its enhancer dose (5.33 nmol/kg) against tetrabenazine-induced learning deficit measured in a shuttle box apparatus was abolished by a 5.84 nmol/kg dose of rasagiline. The selegiline metabolite (−)methamphetamine (10−9 mol/L) also exhibited enhancer activity on [3H]dopamine release. We have concluded that selegiline acts as an MAO-B inhibitor and a dopaminergic enhancer drug, and the latter relates to an agonist effect on TAAR1. In contrast, rasagiline is devoid of enhancer activity but may act as an antagonist on TAAR1.Selegiline and rasagiline are two selective monoamine oxidase B (MAO-B) inhibitors used in the treatment of Parkinson’s disease. In their clinical application, however, differences in L-dopa-sparing potencies have been observed. The aim of this study was to find neurochemical and behavioral explanations for the antiparkinsonian effects of these drugs. We found that selegiline possesses a dopaminergic enhancer effect: it stimulated the electrically induced [ N-(2-propinyl)-2-methyl-1-phenylethylamine), and the compound later became the first known selective B-type monoamine oxidase (MAO) inhibitor dopamine release from the striatum via activation of the trace amine-associated receptor 1 (TAAR1) dopamine tissue content in the striatal slices was 390.59 ± 111.71 kBq/g after the 60 min initial washout period. This content of radioactivity decreased to 232.60 ± 58.03 kBq/g in the following a 75-min-long superfusion. Thus, approximately 40 percent of the striatal tissue’s [3H]dopamine content was released during the superfusion . The resting dopamine release from superfused rat striatal slices methamphetamine resembles the selegiline effect (10−9 mol/L) on the resting and electrical stimulation-induced [3H]dopamine release methamphetamine increased the resting [−5 mol/L B. This eAs 3H]dopamine release measurements, we found that selegiline but not rasagiline exhibits enhancer activity. This observation may serve as neurochemical evidence for previous findings demonstrating the enhancer effect of selegiline in behavioral tests dopamine release without affecting the resting release. Selegiline, however, also increased dopamine release in 10−6 and 10−5 mol/L concentrations; this effect may be related to the inhibition of A-type MAO activity [3H]dopamine deamination in rat striatal slices and found a 76% inhibition at 10−5 mol/L concentration dopamine release by (−)BPAP in the rat striatum dopamine release from rat striatal slices, indicating a lack of enhancer activity of this MAO-B inhibitor. The missing enhancer activity of rasagiline was also shown by behavioral results. While a specific enhancer dose of selegiline reduced the tetrabenazine-induced abolishment of learning ability, rasagiline failed to do so BPAP, which solely possesses enhancer activity without MAO inhibition, may have antiparkinsonian potential .We also confirmed previous findings that (−)methamphetamine, one of the major metabolites of selegiline, possesses a dopaminergic enhancer effect in the striatum ,40. It iAlthough rasagiline has gradually replaced selegiline in the treatment of Parkinson’s disease, clinical comparisons have indicated significant differences in the mode of action of these two drugs. Thus, an early clinical study of rasagiline in Parkinson’s patients failed to demonstrate a decreased need for levodopa, an effect opposite to that of selegiline . A systead libitum. All the experimental procedures were approved by local ethical committees and were carried out in accordance with the NIH Guide for the Care and Use of Laboratory Animals, 8th Edition, 2011.We used male Wistar rats  weighing 180–220 g for our experiments. The animals were housed five per cage in a standard temperature- and humidity-controlled animal facility using a 12 h light/dark cycle . Food and water were available 2/5% CO2) Krebs bicarbonate buffer with a composition of NaCl 118, KCl 4.7, CaCl2 1.25, NaH2PO4 1.2, MgCl2 1.2, NaHCO3 25, glucose 11.5, ascorbic acid 0.3, and Na2EDTA 0.03 mmol/L.The rats were decapitated by a guillotine, and the brains were removed from the skulls. The striatum was dissected as described by Glowinski and Iversen , and str3H]dopamine added to 1.5 mL oxygenated  and preheated (37 °C) Krebs bicarbonate buffer for 30 min [3H]dopamine, and superfused with aerated and preheated Krebs bicarbonate buffer . The flow rate was maintained at 1 mL/min by using a Gilson peristaltic pump . The superfusate was discarded during the first 60 min preperfusion period, then, twenty-five fractions (3 min each) were collected by a fraction collector . Electrical field stimuli  were delivered by a Grass S88 Electrostimulator  to induce vesicular [3H]dopamine release. The drugs were added to the striatal slices between the first and second electrical stimulations, as was indicated in the figure legends.The striatal slices of the rat were incubated in the presence of 10 µCi of [r 30 min . The sup3H]dopamine was calculated as kBq/g tissue.The striatal tissues were collected from the superfusion chambers at the end of the superfusion, weighed, and solubilized in 0.4 mL of Soluene-350. An aliquot (50 µL) was vortex-mixed with a liquid scintillation reagent (5 mL) and subjected to liquid scintillation spectrometry in order to determine the content of radioactivity remaining in the tissues. The tissue content of [3H] radioactivity released from the brain slices, a sample of the collected superfusate (1 mL) was mixed with Soluene-350 liquid scintillation reagent (5 mL) and subjected to liquid scintillation spectrometry. The release of [3H]dopamine was expressed as kBq/g/3 min fraction or as a fractional rate, i.e., a percentage of the amount of radioactivity in the tissue at the time of the release was determined. To calculate the stimulation-evoked [3H]dopamine release, the mean of the basal outflow determined before and after stimulation was subtracted from each sample and summed [For determination of the [d summed .3H]dopamine efflux at rest were expressed by the B2/B1 ratio, i.e., [3H]dopamine outflow in fraction 17  and fraction 3 . The effects of the drugs on the electrical stimulation-evoked [3H]dopamine release were determined with the ratio of [3H]dopamine release measured in response to the 2nd  and 1st  stimulations, i.e., the S2/S1 ratio , saline + tetrabenazine (SAL + TBZ), saline + tetrabenazine + selegiline (SAL + TBZ + SEL), saline + tetrabenazine + rasagiline (SAL + TBZ + RAS), and saline + tetrabenazine + selegiline + rasagiline (SAL + TBZ + SEL + RAS). The tetrabenazine was applied in a dose of 1 mg/kg sc, and the selegiline and rasagiline were injected sc in a dose of 0.001 mg/kg, respectively.t-statistics for two-means, Student’s paired t-test, and one-way ANOVA followed by Dunnett’s test were used for the statistical analysis of the data. The two-way ANOVA and Bonferroni post hoc test were applied for analyzing the shuttle box results. The number of independent determinations was indicated by n, and the data were expressed as the mean ± S.E.M. The level of probability (p) was set at 5% for consideration significance.The Student’s 3H]([3H]dopamine) with specific activity of 27.8 Ci/mmol, Ultima Gold XR liquid scintillation reagent, and Soluene-350 tissue solubilizer were purchased from PerkinElmer Life and Analytical Sciences, Boston, MA, USA. Selegiline ((−)deprenyl, N-methyl-N-(2-propinyl)-2-methyl-1-phenylethylamine), and rasagiline (N-propargyl-(1-indenyl)-ammonium) were gifts from the Fujimoto Pharmaceutical Co., Osaka, Japan. EPPTB -4-pyrrolidin-1-yl-3-trifluoro-methylbenzamide), β-phenylethylamine HCl (PEA), and (R)-(−)-1-aminoindane were purchased from Sigma-Aldrich Chemical Co, Budapest, Hungary. (−)Methamphetamine HCl and (±)methamphetamine HCl were donated by Chinoin Pharmaceuticals, Budapest, Hungary. The tetrabenazine HCl was synthesized by Prof. Dr. Csaba Szantay, Department of Organic Chemistry, Budapest University of Technology and Economics, Budapest, Hungary. All the other chemicals were of analytical grade.Dihydroxyphenylethylamine-3,4[In conclusion, our experiments provide evidence that selegiline possesses a dopaminergic activity enhancer effect in addition to MAO-B enzyme inhibition, and the two effects are independent. In contrast to selegiline, rasagiline solely inhibits MAO-B activity without possessing dopaminergic enhancer activity. We have speculated that the presence of the dopaminergic enhancer activity in the pharmacological spectrum of selegiline may be the basis of the clinical observations reported in the antiparkinsonian effects of selegiline in comparison with rasagiline. Whether the dopaminergic activity enhancer effect of selegiline is responsible for the differences in the levodopa-sparing effect of selegiline and rasagiline is currently being investigated in our laboratory.


Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, and Klebsiella pneumoniae.The aim of this study was to develop a novel amikacin (AMI) delivery system with prolonged release based on composite electrospun nanofibers of PLA supplemented with AMI-loaded Si nanoparticles of different morphology. The resultant materials were characterized in terms of their physical properties . High-Performance Liquid Chromatography was used to determine the AMI content in the liquid fractions obtained from the release study. The results show that nanofibers of fumed silica exhibited an aggregated, highly porous structure, whereas nanofibers of mesoporous silica had a spherical morphology. Both silica nanoparticles had a significant effect on the hydrophilic properties of PLA nanofiber surfaces. The liquid fractions were investigated to gauge the encapsulation efficiency (EE) and loading efficiency (LE) of AMI, demonstrating 66% EE and 52% LE for nanofibers of fumed silica compared to nanofibers of mesoporous silica nanoparticles (52% EE and 12.7% LE). The antibacterial activity of the AMI-loaded nanofibers was determined by the Kirby–Bauer Method. These results demonstrated that the PLA-based silica nanofibers effectively enhanced the antibacterial properties against the  Treating and managing wounds is largely influenced by the type of dressing applied, with various therapeutic possibilities, including protection of the surface, drug delivery, and antibacterial activity ,2,3. EleAn assortment of biodegradable polymers has been utilized to obtain electrospun nanofibers, e.g., poly(ε-caprolactone) , poly delivery system with prolonged, tailored release kinetics, one based on composite electrospun nanofibers of PLA supplemented with AMI-loaded silica nanoparticles of different morphology.AMI is a semisynthetic aminoglycoside antibiotic with broad therapeutic action against Gram-negative bacteria. However, AMI is known to cause adverse effects, including nephrotoxicity and ototoxicity . AnotherIn this work, two types of Si nanoparticles, commercially available fumed silica (SIAM) and fabricated mesoporous silica (SIMES), were investigated for their suitability as supports for AMI immobilization in PLA matrice and for efficient production of PLA-based silica-modified composite nanofibers with optimal diameters. As far as the authors were aware, no study had taken place on the impact exerted by the varied morphology of Si nanoparticles on the morphology of nanofibers in connection with an immobilized bioactive compound in such nanoparticles. The authors researched the hydrophobic properties of the nanofibers, the time-dependent delivery of AMI under simulated physiological conditions, and its antibacterial activity against Gram-positive and Gram-negative bacterial strains in order to gauge the applicability of the nanofibers for wound healing.The following chemicals were utilized in experiments: amikacin disulfate ; L-lactic acid ; Tin(II) 2-ethylhexanoate (Sn(Oct)2; ~95%; Sigma-Aldrich, Schnelldorf, Germany,); solvents—chloroform, acetone, methanol (MeOH) and ethanol ; chloroform ; Fumed silica ; Tetraethyl orthosilicate ; ammonia (25%), dimethylformamide  and ultrapure water—utilized as precursor materials for the synthesis of mesoporous silica nanoparticles; and diiodomethane .S. aureus CCM 4516; E. coli CCM 4517; Enterococcus faecalis CCM 3956; K. pneumoniae CCM 4415; and P. aeruginosa CM 1961.Antibacterial testing was performed on the following bacterial strains : w/w of Sn(Oct)2 added in afterward. The pressure was subsequently reduced to 3 kPa, upon which the reaction continued for 24 h. The product of the polycondensation reaction was precipitated with MeOH and water, filtered off, washed with water and MeOH (10:1), and subsequently dried at 30 °C for 48 h in a vacuum oven (10 kPa). The resultant PLA was characterized by GPC analysis  as follows: three connected LC columns were used—PL gel MIXED-A  + MIXED-B  + MIXED-D ; THF was applied as a mobile phase at the flow rate of 1 mL/min; separation was carried out at 40 °C and the injection volume equaled 100 μL. A refractive index detector and a viscometric detector were employed for the detection. The calibration curves were plotted according to polystyrene standards . The average molar mass or molecular weight (Mw), number average molar mass (Mn), and polydispersity index (Ð = Mw/Mn) of the tested samples were determined from peaks corresponding to the polymer fraction in accordance with the universal calibration method. All data was processed in Cirrus software . This was carried out in accordance with a procedure derived from a method described by Pavelkova et al. . In brie3: 26, C2H5OH: 12, H2O. A typical process took place [2H5OH in 10.4 mL H2O was stirred, into which 3.1 mL of NH3 was added for each set of experiments. This clear solution was supplemented dropwise with 11 mL of TEOS under stirring (400 rpm), and stirring continued for another 4 h at room temperature (25 °C). The resultant solid product was centrifuged  and washed, followed by air drying at 60 °C overnight. Finally, the dried powder was calcined at 550 °C for 6 h at a heating rate of 1 °C min−1.Preparation of SIMES required that the molar composition of the precursors was maintained thus: TEOS: 0.93, NHok place , wherebyTwo types of AMI-loaded nanoparticles formed of either commercially available fumed silica SIAM or fabricated mesoporous silica SIMES were synthesized by the sol-gel method to expedite sustained drug delivery. Incorporating the antibiotic in SIMES and SIAM involved dispersing 500 mg of nanoparticles in 500 mL of AMI aqueous solution (50 mg/mL), and this was subjected to magnetic stirring overnight. The suspension was then centrifuged, and the isolated AMI-loaded silica nanoparticles (SIAM-AMI and SIMES-AMI) were freeze-dried for 48 h. The concentration of AMI in a solid fraction was established by the difference between the initial loading and concentration in the washing solution that was established by HPLC.w/w. Initial quantities of the compounds in the electrospinning mixture are described in Electrospun nanofibers denoted as PLA-SIAM-AMI and PLA-SIMES-AMI were produced from PLA that incorporated the above-mentioned SIMES-AMI and SIAM-AMI (pre-loaded silica nanoparticles with AMI) at a ratio of 1:1 v/v ratio). In order to fabricate fibres that contained AMI, the drug in powder form was dispersed in a polymeric solution that underwent magnetic stirring overnight so as to homogenize the Si nanoparticles in the polymer matrix solution. The final concentration of AMI in resultant nanofibers PLA-SIAM-AMI and PLA-SIMES-AMI were 3.9 and 4.8%, respectively  was used to determine the Si amounts in prepared nanofibers samples. The EDX is based on X-Ray analysis. The samples were analysed in powder form by using the ARL Quant’EDX-RF analyser of Thermo-Fischer Scientific. The analysed samples were placed in a Teflon cup (approximately 3 g) and suitably closed with a special microcellulose film, after which they were inserted into the autosampler. Each sample was analysed twice. Samples were evaluated using the chosen method: Any Sample Helium in the UniQuantX program. The software then evaluated the presence and amount of elements and basic matrix (CHON) in mass percent (% m/m).2 fibre strips taped to a glass slide. Deionized water, ethylene glycol, and diiodomethane at the volume of 2 μL were utilized as testing liquids for the nanofibers. Mean values for contact angle were calculated from 10 separate readings for each such liquid.A SEE System, by Advex Instruments , was employed to gauge the contact angles of samples by the sessile drop method, with the aid of 10.5 cm−1). A heating rate of 10 °C min−1 was applied at intervals across a temperature range of 30 °C to 600 °C. The weight of samples varied between 10 and 12 mg. Values for the organic phase in the carriers were determined by taking into account the extent of weight reduction.The thermal behaviour was studied using thermogravimetric analysis on a Mettler-Toledo TGA/SDTA 851e instrument  under nitrogen flow (20 mL minThe specific surface area and pore size distribution of the silica nanoparticles and those loaded with AMI were derived from recordings of nitrogen adsorption–desorption isotherms made on a Micrometrics Brunauer–Emmett–Teller (BET)-surface device . The pore size distribution was ascertained from the desorption branches of the isotherms by following the Barrett–Joyner–Halenda (BJH) method.SEM micrographs of the prepared formulations coated with a sputtered gold/palladium layer  were obtained on Nova NanoSEM 450 scanning electron microscope set to 5 kV. Mean values for fibre diameter and related distribution were estimated from SEM images, facilitated by measuring the diameters of fifty randomly selected nanofibers with the aid of image analysis software .2 were immersed in 10 mL of PBS in capped glass flasks and then incubated at 37 °C in a thermostatically controlled orbital shaker set to 100 rpm. At predefined time intervals, 1 mL of the liquid fraction of each sample was collected, and the rest of release medium was completely removed and substituted with the same amount of fresh PBS. Sink condition tests were carried out, and the AMI content of each sample was determined by High-Performance Liquid Chromatography (HPLC) described below. Following the release studies, the total amount of AMI present was calculated by dissolving samples in 1 mL of chloroform, and any remaining AMI was extracted by PBS. Release studies were performed in triplicate for each type of fibre.The release kinetics of AMI were determined by applying a phosphate-buffered saline solution  as the release medium. Samples measuring 14 cm3BO3/100 mL, pH = 11), 3 µL of OPA* reagent, 1 µL of 1 M acetic acid. The separation after derivatization was performed on an XSELECT CSH C18 5 µm column  equipped with a security guard column  at 30 °C. A mixture of 100 mM Acetate buffer  and HPLC grade Acetonitrile  was used as mobile phase  at a flow rate of 0.4 mL/min; FLD detection was performed at λexcitation = 330 nm and λemission = 440 nm.AMI was determined by in-needle derivatization with o-phthaldialdehyde (OPA) reagent and FLD detection . In-needle derivatization followed a specific injection schedule as follows: 1 µL of the original sample solution was gradually mixed with the solutions in the needle: 5 µL of borate buffer  were prepared by diluting the stock solution of AMI (1 mg/mL) with PBS solution (pH = 7.4). An external calibration method was used to determine the amount of the analyte. The limits of detection and quantification were determined as LOD = 0.20 µg/mL and LOQ = 0.60 µg/mL, respectively. The obtained calibration curve was linear with a coefficient of determination R1 is the total amount of AMI loading in the nanoparticles (mg), C2 denotes the amount of excess AMI in the waste solution after the material had been prepared (mg), and Wm constitutes the weight of the dry mass of the prepared material (mg).The liquid fractions of the samples were also investigated to gauge encapsulation efficiency (EE) and loading efficiency (LE) in percent, as per the following equations (Equation (1): EE; Equation (2): LE):Staphylococcus aureus CCM 4516, Escherichia coli CCM 4517, Enterococcus faecalis CCM 3956, Klebsiella pmeumoniae CCM 4415 and Pseudomonas aeruginosa CM 1961 was determined by the disk diffusion test (Kirby–Bauer Method), in line with the European Committee on Antimicrobial Susceptibility Testing (EUCAST) [6–107 cfu/mL. The samples of prepared nanofibers were cut into round pieces ). The AMI was used in the same quantity. The samples were formed into two piles in one Petri dish, and another inoculated with Mueller–Hinton agar placed on top (performed in triplicate), then the combined plates were incubated at 35 °C for 18 to 24 h. Following incubation, the width of the zone of inhibition for each sample was measured to the nearest millimetre on a SCAN 500 inhibition zone reader (version 8.2.0.0). Nanofibers without AMI were also tested as the control. Each type of fibre was tested in triplicate.Such activity of the AMI-loaded nanofibers against (EUCAST) . The conSurface properties were investigated by the BET method. 3-type hysteresis loops at high relative pressure, according to IUPAC classifications, which is a characteristic of mesoporous materials with great uniformity in size distribution [o region from 0.8 to 1.0, indicated that the materials possessed a well-defined and regular array of mesopores caused by gaps between the nanofibers. The PLA-SIMES-AMI sample exhibited a hysteresis loop at low relative pressure, connected with the availability of mesopores on the surfaces of the Si nanoparticles, themselves available on the outer sides of the nanofibers. The structured data of each mesoporous material by BET showed that the PLA-Si-based nanofibers had associated high surface areas  are displayed in w/w) for SIAM and 40% (w/w) for SIMES, as presented in EE and LE were calculated via Equations (1) and (2), which revealed the Si nanoparticles had a high loading capacity due to their high surface area . Values p < 0.05) than formulations with Si nanoparticles. The diameter distributions of the AMI-loaded electrospun nanofibers are presented in The morphology of SIAM, SIMES, and electrospun nanofibers was observed by SEM. The EDXRF method was applied to gauge the content of CHO and Si (%) incorporated in the AMI-loaded nanofibers. Taking into consideration the extent of Si present from results obtained for the loading of the Si particles, this method revealed that materials containing SIMES demonstrated a slightly higher presence of Si nanoparticles see . As for The literature refers to pure PLA composite fibres of a nano/microstructure capable of prolonging the release of several drugs ,33,52 inS. aureus, E. coli, P. aeruginosa, and K. pneumoniae, possibly through the more rapid release of the antibiotic from their open-pore structure.The fabricated nanofibers were assumed to possess enhanced antibacterial ability against Gram-positive and Gram-negative bacterial strains; hence these properties of the PLA-Si nanofibers were studied in detail. The disk diffusion test was carried out to investigate the antibacterial ability of AMI-loaded fibre materials against selected bacterial strains, which are the most common causative organisms associated with wound infection . As descThe aim of this study was to create a novel material with antibacterial properties to counteract wound infection. In this context, two types of AMI-loaded nanoparticles formed of either commercially available fumed silica SIAM or fabricated mesoporous silica SIMES were incorporated in electrospun PLA nanofibers to facilitate prolonged, sustained release of the antibiotic. The results by BET showed that the PLA-Si-based nanofibers had an associated high surface area, large pore volumes, and suitable pore sizes, marking them out as potentially suitable as drug delivery carriers. The incorporation of the silica nanoparticles changed the hydrophobicity of the nanofibers to an extent, which in the context of the intended application, would advantageously curtail inflammatory reactions in living tissue. It was observed that supplementing silica nanoparticles with the AMI and embedding them in nanofibers by electrospinning gave rise to drug-loaded electrospun nanofibers with different release profiles. Experiments revealed that the presence of Si nanoparticles also enhanced the antibacterial properties of the given agent. Since enriching the PLA nanofibers with Si nanoparticles brought about a dramatic increase in antibacterial ability, it would be possible to load the AMI at a limited concentration as a consequence of the Si nanoparticles being so effective against the tested organisms. This antibacterial experiment found that prepared PLA-Si-based nanofibers inherently exerted a strong antibacterial effect against the most common causative Gram-positive and Gram-negative bacteria associated with wound infection, making these nanofibers applicable for wound treatment.