Cell sensitivity, non-linearity and inverse effects

It has been claimed that the homeopathic principle of 'similarity' (or 'similia') and the use of individualized remedies in extremely low doses conflicts with scientific laws, but this opinion can be disputed on the basis of recent scientific advances. Several mechanisms to explain the responsiveness of cells to ultra-low doses and the similarity as inversion of drug effects, have again been suggested in the framework of hormesis and modern paradoxical pharmacology. Low doses or high dilutions of a drug interact only with the enhanced sensitivities of regulatory systems, functioning as minute harmful stimuli to trigger specific compensatory healing reactions. Here we review hypotheses about homeopathic drug action at cellular and molecular levels, and present a new conceptual model of the principle of similarity based on allosteric drug action. While many common drugs act through orthostatic chemical interactions aimed at blocking undesired activities of enzymes or receptors, allosteric interactions are associated with dynamic conformational changes and functional transitions in target proteins, which enhance or inhibit specific cellular actions in normal or disease states. The concept of allostery and the way it controls physiological activities can be broadened to include diluted/dynamized compounds, and may constitute a working hypothesis for the study of molecular mechanisms underlying the inversion of drug effects

Study of the action mechanisms of Arnica montana effects on macrophages

Objective: To test the effect of Arnica montana (Arnica m.) on human macrophages. Method: The human monocytic leukaemia cell line THP-1 was cultured and differentiated in mature macrophages with PMA and other differentiating agents. Macrophages were exposed to Arnica m. homeopathic dilutions (2c, 3c, 5c, 9c and 15c) or Control solvent. Total RNA was isolated and sequenced to perform quantitative evaluation. Results: Screening sorted out with a list of 20 genes that were significantly changed by Arnica m. 2c treatment: 7 up-regulated and 13 down-regulated. Most notably, a clearly up-regulated function concerned the proteinaceous extracellular matrix (ECM), including genes HSPG2, FBN2, FN1 (

Biological activity of interferon gamma and lipopolysaccharide on the nitric oxide production in C6 astroglioma cells and some unexpected effects of potentization

Background: A proinflammatory environment is a hallmark of several neurodegenerative diseases where astrocyte involvement is also well established. Astrocytes and microglia in central nervous system are mainly involved in the release of cytokines, oxygen free radicals and nitric oxide (NO). Several studies on C6 astroglioma cells, a widely used in vitro model for these events, demonstrated that co-stimulation of this cell line with bacterial lipopolysaccharide (LPS) and interferon gamma (IFN-) induces a synergistic nitric oxide synthase (iNOS) expression.1 In our laboratory we are using this versatile cell model in order to carefully investigate dose-response effects of various putative agonists or inhibitors and to assess the possible changes provoked in those agents by different procedures of dilution and succussion (agitation) (potentization according to the homeopathic terminology)

Testing Homeopathy in Mouse Emotional Response Models: Pooled Data Analysis of Two Series of Studies

Two previous investigations were performed to assess the activity of Gelsemium sempervirens (Gelsemium s.) in mice, using emotional response models. These two series are pooled and analysed here. Gelsemium s. in various homeopathic centesimal dilutions/dynamizations (4C, 5C, 7C, 9C, and 30C), a placebo (solvent vehicle), and the reference drugs diazepam (1 mg/kg body weight) or buspirone (5 mg/kg body weight) were delivered intraperitoneally to groups of albino CD1 mice, and their effects on animal behaviour were assessed by the light-dark (LD) choice test and the open-field (OF) exploration test. Up to 14 separate replications were carried out in fully blind and randomised conditions. Pooled analysis demonstrated highly significant effects of Gelsemium s. 5C, 7C, and 30C on the OF parameter “time spent in central area” and of Gelsemium s. 5C, 9C, and 30C on the LD parameters “time spent in lit area” and “number of light-dark transitions,” without any sedative action or adverse effects on locomotion. This pooled data analysis confirms and reinforces the evidence that Gelsemium s. regulates emotional responses and behaviour of laboratory mice in a nonlinear fashion with dilution/dynamization

Serum Exosomal microRNA-21, 222 and 124-3p as Noninvasive Predictive Biomarkers in Newly Diagnosed High-Grade Gliomas: A Prospective Study

Background: High-grade gliomas (HGG) are malignant brain tumors associated with frequent recurrent disease. Clinical management of HGG patients is currently devoid of blood biomarkers for early diagnosis, monitoring therapeutic effects and predicting recurrence. Different circulating miRNAs, both free and associated with exosomes, are described in patients with HGG. We previously identified miR-21, miR-222 and miR-124-3p purified from serum exosomes as molecular signature to help pre-operative clinical diagnosis and grading of gliomas. The aim of the present study was to verify this signature as a tool to assess the effect of treatment and for the early identification of progression in newly diagnosed HGG patients. Material and Methods: Major inclusion criteria were newly diagnosed, histologically confirmed HGG patients, no prior chemotherapy, ECOG PS 0-2 and patients scheduled for radiochemotherapy with temozolomide as first-line treatment after surgery. RANO criteria were used for response assessment. Serum was collected at baseline and subsequently at each neuroradiological assessment. mir-21, -222 and -124-3p expression in serum exosomes was measured in all samples. Results: A total number of 57 patients were enrolled; 41 were male, 52 with glioblastoma and 5 with anaplastic astrocytoma; 18 received radical surgery. HGG patients with higher exosomal miRNA expression displayed a statistically significant lower progression-free survival and overall survival. Increased expression of miR-21, -222 and -124-3p during post-operative follow-up was associated with HGG progression. Conclusions: These data indicate that miR-21, -222 and -124-3p in serum exosomes may be useful molecular biomarkers for complementing clinical evaluation of early tumor progression during post-surgical therapy in patients with HGG

GM1 as adjuvant of innovative therapies for cystic fibrosis disease

Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein is expressed at the apical plasma membrane (PM) of different epithelial cells. The most common mutation responsible for the onset of cystic fibrosis (CF), F508del, inhibits the biosynthesis and transport of the protein at PM, and also presents gating and stability defects of the membrane anion channel upon its rescue by the use of correctors and potentiators. This prompted a multiple drug strategy for F508delCFTR aimed simultaneously at its rescue, functional potentiation and PM stabilization. Since ganglioside GM1 is involved in the functional stabilization of transmembrane proteins, we investigated its role as an adjuvant to increase the effectiveness of CFTR modulators. According to our results, we found that GM1 resides in the same PM microenvironment as CFTR. In CF cells, the expression of the mutated channel is accompanied by a decrease in the PM GM1 content. Interestingly, by the exogenous administration of GM1, it becomes a component of the PM, reducing the destabilizing effect of the potentiator VX-770 on rescued CFTR protein expression/function and improving its stabilization. This evidence could represent a starting point for developing innovative therapeutic strategies based on the co-administration of GM1, correctors and potentiators, with the aim of improving F508del CFTR function

Applicazioni delle biotecnologie in microbiologia clinica

La biologia molecolare negli ultimi decenni ha rivoluzionato il panorama della ricerca scientifica e della diagnostica, permettendo di rendere visibili e di quantificare \u201centit\ue0\u201d delle quali, in un passato non molto lontano, si ignorava persino l\u2019esistenza. Se fino agli anni \u201950 si parlava di presunte proteine in grado di trasmettere l\u2019informazione genetica, ora si conoscono esattamente le molecole responsabili del perpetuarsi della vita: l\u2019acido deossiribonucleico (DNA) e l\u2019acido ribonucleico (RNA). In un susseguirsi incessante di scoperte l\u2019uomo prende pieno possesso della chimica, della biologia e della struttura degli acidi nucleici. L\u2019essenza di ogni forma di vita diventa decifrabile in pochi e semplici elementi: le basi azotate, che costituiscono una catena fitta di informazioni e legami chimici, codificano, di tre in tre, gli aminoacidi da inglobare nelle nascenti proteine. \uc8 la stele di Rosetta per decriptare il codice genetico: il gene da entit\ue0 astratta prende forma, sostanza e dimora. Il processo di \u201cmodernizzazione\u201d della scienza della vita \ue8 cominciato: nasce la biologia molecolare che in pochi anni rivoluziona l\u2019indagine scientifica in campo medico, farmaceutico, alimentare, ambientale\u2026 Dopo l\u2019introduzione della reazione di amplificazione a catena (PCR) del DNA, avvenuta negli anni \u201980, il codice genetico della vita diventa \u201cvisibile\u201d e a \u201cportata d\u2019uomo\u201d: una singola molecola viene replicata in modo esponenziale milioni di volte, milioni di copie di un gene sono disponibili per essere studiate, comprese e clonate. I geni di interesse vengono inseriti in vettori ricombinanti, a loro volta ospiti di quelli che oggi chiamiamo organismi geneticamente modificati (OGM). In tutti i laboratori del mondo prendono vita nuove entit\ue0 biologiche, con geni ancestrali frutto di una selezione milionaria, ma in combinazioni che solo l\u2019uomo ha saputo creare. E non solo. Nel XXI secolo l\u2019uomo riesce anche a sequenziare il proprio genoma, a scrivere il vocabolario della sua storia biologica, pur senza comprendere pienamente il significato di ogni \u201cparola\u201d. Ma, indipendentemente dalla comprensione di queste \u201cparole\u201d, \ue8 opportuno chiedersi quanto il contesto ambientale, gli eventi contingenti, limitino e condizionino la totipotenza del nostro codice genetico. Sotto questo profilo, in una visione deterministica, l\u2019uomo potrebbe presuntuosamente credere di aver raggiunto lo scopo, quello di prevedere l\u2019insorgere di malattie o patologie, quello di modellare la vita secondo i propri gusti e desideri\u2026 Ma la persuasione di ci\uf2 \ue8 lontanissima dalla realt\ue0 cosmica, policromatica, mutevole di cui siamo pervasi. Le conoscenze di biologia molecolare acquisite in questi ultimi anni ci possono aiutare ad indagare, capillarmente, i meccanismi alla base di molte malattie, ad accelerare i tempi per una diagnosi, a disegnare farmaci personalizzati, a migliorare le caratteristiche di un prodotto alimentare o di un vegetale necessario per l\u2019alimentazione umana e animale, ma non potranno mai prevedere, secondo una visione riduzionista della scienza, il nostro destino biologico.not availabl

Response to Comment on Olioso D. et al. "Effect of Aerobic and Resistance Training on Circulating Micro-RNA Expression Profile in Subjects with Type 2 Diabetes. J Clin Endocrinol Metab 2019; 104(4):1119-1130" by Chirumbolo S

[no abstract available

Biological activity of interferon gamma and lipopolysaccharide on the nitric oxide production in C6 astroglioma cells and some unexpected effects of potentization

Background: The search for new therapeutic approaches with fewer side effects and better treatment efficacy to the Chagas Disease has been a major challenge. Aim: To evaluate the effects of Kalium causticum, Conium maculatum, and Lycopodium clavatum 13 cH in mice inoculated with the Y strain of Trypanosoma cruzi. Materials and methods: In a blind, controlled, randomized study, 102 male Swiss mice, eight weeks old, were inoculated with 1,400 trypomastigotes of the Y strain of T. cruzi and distributed into the following groups: CI (treated with 7% hydroalcoholic solution), Ca (treated with Kalium causticum 13cH), Co (treated with Conium maculatum 13cH), and Ly (treated with Lycopodium clavatum 13cH). The medicines were selected by three homeopaths using Lince Expert System Software (Albuquerque, NM, USA), considering the behavioral characteristics of the mice. The treatments were performed 48 hours before and 48, 96, and 144 hours after infection [1]. The following parameters were evaluated: infectivity, prepatent period, parasitemia peak, total parasitemia, tissue tropism, inflammatory infiltrate, and survival. Results: The prepatent period was greater in the Ly group than in the CI group (p = 0.02). The number of trypomastigotes on the 8th day after infection was lower in the Ca group than in the CI group (p < 0.05). Total parasitemia was significantly lower in the Ca, Co, and Ly groups than in the CI group. On the 12th day after infection, the Ca, Co, and Ly groups had fewer nests of amastigotes and amastigotes/nest in the heart than the CI group (p < 0.05) (Figure-I). A decrease in the number of nests and amastigotes in the intestine were observed in the Ly group compared with the CI group (p < 0.05). In the liver (day 12), Ly significantly prevented the formation of inflammatory foci compared with the other groups. In muscle, Co and Ly decreased the formation of inflammatory foci compared with CI (p < 0.05). Ly afforded greater animal survival compared with CI, Ca, and Co (p < 0.05). The animals in the Co group died prematurely compared with the CI group (p = 0.031). (Figure-II) Conclusion: All of the experimental homeopathic medications with 13cH dynamization studied herein reduced the parasite peak and total parasitemia. Ly had significantly more benefits in the treatment of mice infected with T. cruzi, reducing the number of blood parasites, amastigotes nests in tissue and the number of amastigotes per nest, resulting in the increasing animal survival. The data may contribute to changes in management strategies in individuals with Chagas disease

Arnica Montana effects on gene expression in a human macrophage cell line. Evaluation by quantitative Real-Time PCR

Background: Arnica montana is a popular traditional remedy widely used in complementary medicine, also for its wound healing properties. Despite its acknowledged action in clinical settings at various doses, the molecular aspects relating to how Arnica montana promotes wound healing remain to be elucidated. To fill this gap, we evaluated the whole plant extract, in a wide range of dilutions, in THP-1 human cells, differentiated into mature macrophages and into an alternative IL-4-activated phenotype involved in tissue remodelling and healing. Methods: Real-time quantitative Reverse Transcription Polymerase Chain Reaction (PCR) analysis was used To study the changes in the expression of a customized panel of key genes, mainly cytokines, receptors and transcription factors. Results: On macrophages differentiated towards the wound healing phenotype, Arnica montana affected the expression of several genes. In particular CXC chemokine ligand 1 (CXCL1), coding for a chief chemo- kine, exhibited the most consistent increase of expression, while also CXC chemokine ligand 2 (CXCL2), Interleukin8 (IL8) and bone morphogenetic protein (BMP2) were slightly up-regulated, suggesting a positive influence of Arnica montana on neutrophil recruitment and on angiogenesis. MMP1, coding for a metalloproteinase capable of cleaving extracellular matrix substrates, was down-regulated. Most results showed non-linearity of the dose-effect relationship. Conclusions: This exploratory study provides new insights into the cellular and molecular mechanisms of action of Arnica montana as a promoter of healing, since some of the genes it modifies are key regulators of tissue remodelling, inflammation and chemotaxis