Early pathways of maternal mentalization: Associations with child development in the FinnBrain birth cohort study.

Parental mentalization refers to a parents' capacity and interest to consider the individual experience and mental state underlying the behaviors of the child. Higher mentalization is considered a key aspect for parental sensitivity in interaction, fostering child's socioemotional and self-regulatory development. Yet, previous studies have not examined the dynamic pathways through which the maternal mentalization may develop, nor their effects on child development. Thus, in the current person-oriented study, first, we identify distinct profiles and longitudinal trajectories of maternal mentalization from pregnancy to child's 2 years of age. Second, we test how the profiles and trajectories associate with children's internalizing and externalizing problems, social-emotional competence and effortful control at the age of 2 years. Third, we examine how the profiles and trajectories associate with contextual demographic and child related. The substudy was part of the FinnBrain Birth Cohort and included families from general population (n = 2,687). Mothers reported their parental reflective functioning (PRF) at late pregnancy, 6 months and 2 years of child's age. Both mothers (n = 1,437) and fathers (n = 715) reported the developmental child outcomes at the child's age of 2 years. Latent Profile Analysis and Latent Transition Analysis were used to identify PRF profiles and trajectories. The results showed decreasing heterogeneity in PRF from pregnancy to child's age of 6 months and 2 years (i.e., four, three and two latent classes, respectively). Most mothers progressed towards high PRF over time. Second, the profiles and trajectories depicting high PRF associated with child high social-emotional competence at the age of 2 years, yet no clear positive effects were found on child's problems and effortful control. The group of mixed PRF trajectories showed strongest association with child's internalizing and externalizing problems. Finally, there were theoretically meaningful associations between the PRF trajectories and both the contextual (e.g., parity) and child related (e.g., infant temperament) factors. This was the first study to explore the early unfolding of maternal mentalization. The results are discussed in relation with the potential mechanisms accounting for child development and with the nature and limitations of self-reported parental mentalization.

The efficacy of probiotics and/or n-3 long-chain polyunsaturated fatty acids intervention on maternal prenatal and postnatal depressive and anxiety symptoms among overweight and obese women.

BACKGROUND: Maternal depression and anxiety may endanger well-being of both mother and child. We investigated the efficacy of probiotics and/or fish oil (FO) in modifying pre- and postnatal depressive and anxiety symptoms. Symptom trajectories were identified and the influence of lifestyle factors on symptoms was evaluated. METHODS: Overweight women (n = 439) were randomized to intervention groups (probiotics+FO, probiotics+placebo, FO+placebo, placebo+placebo) from early pregnancy until six months postpartum, and assessed for depressive and anxiety symptoms with Edinburgh Postnatal Depression Scale (EPDS) and Anxiety subscale of Symptoms Checklist (SCL-90) at early and late pregnancy and three, six and 12 months postpartum. Latent growth mixture modeling was used to model the symptom courses. Dietary quality and physical activity were assessed with validated indices. RESULTS: Symptom scores were generally low. Statistically significant intervention effect was seen during pregnancy (p = 0.017): EPDS scores increased (by 1.11 points) in the FO+probiotics group and decreased (by 0.85 points) in the FO+placebo group. At 12 months postpartum, FO+placebo group had lower EPDS scores compared to probiotics+placebo group (p = 0.039). No differences in SCL scores were seen in response to the intervention. Irrespective of the intervention, three depressive and two anxiety symptoms trajectories were identified. Dietary quality correlated negatively with depressive symptoms in early pregnancy and six months postpartum and with anxiety symptoms in early pregnancy. Perinatal events including mother-reported colic were related to symptoms. LIMITATIONS: Secondary outcomes of the primary trial. CONCLUSIONS: Intervention had a modest impact on depressive symptoms. Diet and obstetric events were associated with depressive and anxiety symptoms.

Welding fume nanoparticles from solid and flux-cored wires: Solubility, toxicity, and role of fluorides.

Welding fume particles are hazardous. Their toxicity likely depends on their composition and reactivity. This study aimed at exploring the role of sodium or other fluorides (NaF), which are intentionally added to flux-cored wire electrodes for stainless steel welding, on the solubility (in phosphate buffered saline) and toxicity of the generated welding fume particles. A multi-analytical particle characterization approach along with in-vitro cell assays was undertaken. The release of Cr(VI) and Mn from the particles was tested as a function of fluoride solution concentration. The welding fume particles containing NaF released significantly higher amounts of Cr(VI) compared with solid wire reference fumes, which was associated with increased cytotoxicity and genotoxicity in-vitro. No crystalline Na or potassium (K) containing chromates were observed. Cr(VI) was incorporated in an amorphous mixed oxide. Solution-added fluorides did not increase the solubility of Cr(VI), but contributed to a reduced Mn release from both solid and flux-cored wire fume particles and the reduction of Cr(VI) release from solid wire fume particles. Chemical speciation modeling suggested that metal fluoride complexes were not formed. The presence of NaF in the welding electrodes did not have any direct, but possibly an indirect, role in the Cr(VI) solubility of welding fumes.

A variation in the infant oxytocin receptor gene modulates infant hippocampal volumes in association with sex and prenatal maternal anxiety.

Genetic variants in the oxytocin receptor (OTR) have been linked to distinct social phenotypes, psychiatric disorders and brain volume alterations in adults. However, to date, it is unknown how OTR genotype shapes prenatal brain development and whether it interacts with maternal prenatal environmental risk factors on infant brain volumes. In 105 Finnish mother-infant dyads (44 female, 11-54 days old), the association of offspring OTR genotype rs53576 and its interaction with prenatal maternal anxiety (revised Symptom Checklist 90, gestational weeks 14, 24, 34) on infant bilateral amygdalar, hippocampal and caudate volumes were probed. A sex-specific main effect of rs53576 on infant left hippocampal volumes was observed. In boys compared to girls, left hippocampal volumes were significantly larger in GG-homozygotes compared to A-allele carriers. Furthermore, genotype rs53576 and prenatal maternal anxiety significantly interacted on right hippocampal volumes irrespective of sex. Higher maternal anxiety was associated both with larger hippocampal volumes in A-allele carriers than GG-homozygotes, and, though statistically weak, also with smaller right caudate volumes in GG-homozygotes than A-allele carriers. Our study results suggest that OTR genotype enhances hippocampal neurogenesis in male GG-homozygotes. Further, prenatal maternal anxiety might induce brain alterations that render GG-homozygotes compared to A-allele carriers more vulnerable to depression.

Partial Support for an Interaction Between a Polygenic Risk Score for Major Depressive Disorder and Prenatal Maternal Depressive Symptoms on Infant Right Amygdalar Volumes.

Psychiatric disease susceptibility partly originates prenatally and is shaped by an interplay of genetic and environmental risk factors. A recent study has provided preliminary evidence that an offspring polygenic risk score for major depressive disorder (PRS-MDD), based on European ancestry, interacts with prenatal maternal depressive symptoms (GxE) on neonatal right amygdalar (US and Asian cohort) and hippocampal volumes (Asian cohort). However, to date, this GxE interplay has only been addressed by one study and is yet unknown for a European ancestry sample. We investigated in 105 Finnish mother-infant dyads (44 female, 11-54 days old) how offspring PRS-MDD interacts with prenatal maternal depressive symptoms (Edinburgh Postnatal Depression Scale, gestational weeks 14, 24, 34) on infant amygdalar and hippocampal volumes. We found a GxE effect on right amygdalar volumes, significant in the main analysis, but nonsignificant after multiple comparison correction and some of the control analyses, whose direction paralleled the US cohort findings. Additional exploratory analyses suggested a sex-specific GxE effect on right hippocampal volumes. Our study is the first to provide support, though statistically weak, for an interplay of offspring PRS-MDD and prenatal maternal depressive symptoms on infant limbic brain volumes in a cohort matched to the PRS-MDD discovery sample.

Presence of CD3+ and CD79a+ Lymphocytes in the Pituitary Gland of Dogs at Post-mortem Examination.

Hypophysitis has been reported occasionally in dogs, with most cases resembling primary lymphocytic hypophysitis in man. Although it is generally assumed that lymphocytes are not present normally in the canine pituitary gland, few studies have investigated this hypothesis. However, lymphocytes are recognized in the pituitary gland of people and horses without signs of pituitary disease. It is unknown to what degree lymphocyte infiltration of the pituitary gland might occur as an incidental finding in dogs. The aim of the present study was to investigate the presence and distribution of lymphocytes in the pituitary gland of dogs without clinical suspicion of pituitary disease. Twenty dogs were subjected to routine necropsy examination. Formalin-fixed and paraffin wax-embedded sections of pituitary were stained with haematoxylin and eosin (HE) or subjected to immunohistochemistry (IHC) using primary antibodies specific for the T-cell marker CD3 and the B-cell marker CD79a. The number of CD3+ and CD79a+ cells per area unit (CPA) was determined for different pituitary regions. Two dogs had extensive neoplastic lesions in the pituitary gland and were excluded from analysis. In the remaining 18 dogs, occasional scattered CD3+ cells were found in the pituitary gland. There was a significant difference in CD3+ CPA between pituitary regions (P = 0.001). The highest CD3+ CPA was found in the pars tuberalis (median 41.3 cells/mm2, interquartile range 20.9-50.5 cells/mm2). In six of the 18 dogs (33%), CD79a+ cells were detected in small number (median total cell number 0 cells/section, interquartile range 0-1.0 cells/section). This study shows that T cell, and fewer B cells, may be found in the pituitary gland of dogs without clinical suspicion of pituitary disease. Regional difference in T-cell density, with the highest CD3+ CPA in the pars tuberalis, may imply regional immunoregulatory functions in the canine pituitary gland.

Associations of age and sex with brain volumes and asymmetry in 2-5-week-old infants.

Information on normal brain structure and development facilitates the recognition of abnormal developmental trajectories and thus needs to be studied in more detail. We imaged 68 healthy infants aged 2-5 weeks with high-resolution structural MRI (magnetic resonance imaging) and investigated hemispheric asymmetry as well as the associations of various total and lobar brain volumes with infant age and sex. We found similar hemispheric asymmetry in both sexes, seen as larger volumes of the right temporal lobe, and of the left parietal and occipital lobes. The degree of asymmetry did not vary with age. Regardless of controlling for gestational age, gray and white matter had different age-related growth patterns. This is a reflection of gray matter growth being greater in the first years, while white matter growth extends into early adulthood. Sex-dependent differences were seen in gray matter as larger regional absolute volumes in males and as larger regional relative volumes in females. Our results are in line with previous studies and expand our understanding of infant brain development.

Nature of magnetization and lateral spin-orbit interaction in gated semiconductor nanowires.

Semiconductor nanowires are interesting candidates for realization of spintronics devices. In this paper we study electronic states and effects of lateral spin-orbit coupling (LSOC) in a one-dimensional asymmetrically biased nanowire using the Hartree-Fock method with Dirac interaction. We have shown that spin polarization can be triggered by LSOC at finite source-drain bias,as a result of numerical noise representing a random magnetic field due to wiring or a random background magnetic field by Earth magnetic field, for instance. The electrons spontaneously arrange into spin rows in the wire due to electron interactions leading to a finite spin polarization. The direction of polarization is, however, random at zero source-drain bias. We have found that LSOC has an effect on orientation of spin rows only in the case when source-drain bias is applied.

Behavioral testing and litter effects in the rabbit.

BACKGROUND: Behavioral testing provides an essential approach in further developing our understanding of brain structure and function. The aim of our study was to outline a more expanded approach to cognition- and anxiety-related behavior in the rabbit. METHODS: Twenty-one 70-day old rabbits (13 female, 8 male) were exposed to open field test, dark-light box test and object recognition testing with variations in inter-trial-interval, olfactory recognition and object location testing. Independent T-tests were used to compare data by individual baseline characteristics, i.e. birth weight, weight at testing, sex, litter #, litter size. RESULTS: In the open field test, median time spent in the center was 3.64 s (0.84-41.36) for the 9 rabbits who entered the center; median distance moved in the arena was 874.42 cm (54.20-3444.83). In the dark light box test, 12 rabbits entered the light compartment. In the object recognition task, rabbits spent significantly less time exploring the familiar object compared to the novel (0.40 s [0-2.8] vs. 3.17 s [1.30-32.69]; P = 0.003) when using a 30-min inter-trial interval, as well with a 90-min inter-trial interval: 0.87 s [0-7.8] vs. 7.65 s [0-37.6] (P = 0.008). However, recognition was lost when using a 24-h inter-trial interval (time spent exploring the familiar object: 3.33 [0-10.90]; novel object:3.87 [1.15-48.53]; n.s). In the object location task and in olfactory object recognition task, median discrimination indexes were 0.69 (-1 to 1) and 0.37 (-0.38 to 0.78) respectively, higher than level expected by chance (P < 0.001). Litter size >3 during the neonatal period was associated with increased explorative behavior in the dark light box test (P = 0.046) and in the visual object recognition task (P = 0.005), whereas body weight and sex were not. CONCLUSIONS: Settings and outcome measures for multiple behavioral tests, providing reference values and considerations for future developmental studies are reported. Discrimination and memory in the rabbit appear to relate to litter characteristics, although a larger sample size is needed to confirm our findings.

Lipoprotein modifications by gingipains of Porphyromonas gingivalis.

BACKGROUND AND OBJECTIVE: Several studies have shown an association between periodontitis and cardiovascular disease (CVD). Atherosclerosis is the major cause of CVD, and a key event in the development of atherosclerosis is accumulation of lipoproteins within the arterial wall. Bacteria are the primary etiologic agents in periodontitis and Porphyromonas gingivalis is the major pathogen in the disease. Several studies support a role of modified low-density lipoprotein (LDL) in atherogenesis; however, the pathogenic stimuli that induce the changes and the mechanisms by which this occur are unknown. This study aims to identify alterations in plasma lipoproteins induced by the periodontopathic species of bacterium, P. gingivalis, in vitro. MATERIAL AND METHODS: Plasma lipoproteins were isolated from whole blood treated with wild-type and gingipain-mutant (lacking either the Rgp- or Kgp gingipains) P. gingivalis by density/gradient-ultracentrifugation and were studied using 2-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ionization mass spectrometry. Porphyromonas gingivalis-induced lipid peroxidation and antioxidant levels were measured by thiobarbituric acid-reactive substances and antioxidant assay kits, respectively, and lumiaggregometry was used for measurement of reactive oxygen species (ROS) and aggregation. RESULTS: Porphyromonas gingivalis exerted substantial proteolytic effects on the lipoproteins. The Rgp gingipains were responsible for producing 2 apoE fragments, as well as 2 apoB-100 fragments, in LDL, and the Kgp gingipain produced an unidentified fragment in high-density lipoproteins. Porphyromonas gingivalis and its different gingipain variants induced ROS and consumed antioxidants. Both the Rgp and Kgp gingipains were involved in inducing lipid peroxidation. CONCLUSION: Porphyromonas gingivalis has the potential to change the expression of lipoproteins in blood, which may represent a crucial link between periodontitis and CVD.

Illuminating the clinical significance of alexithymia subtypes: A cluster analysis of alexithymic traits and psychiatric symptoms.

BACKGROUND: Alexithymia is a personality construct involving difficulties identifying and verbalizing feelings, and an externally oriented thinking style. There is preliminary evidence for alexithymia subtypes that may carry different risk profiles for psychiatric illness. The aim of this study was to gain support for the existence of alexithymia subtypes and further characterize their clinical relevance. METHODS: To identify possible subtypes, a cluster analysis was conducted for individuals with high alexithymic traits (N=113). Current depressive and anxiety symptoms, self-reported psychiatric medical history, and self-reported early life adversity were compared between subtypes. The cluster analysis was replicated with the low (N=2471) and moderate (N=290) alexithymia groups. RESULTS: We identified two alexithymia subtypes. Compared to type A, type B alexithymia was associated with higher levels of difficulties in identifying feelings, and was more strongly associated with current depressive (Cohen's d=0.77, p<0.001) and anxiety symptoms (Cohen's d=0.82, p<0.001), and self-reported early life adversity (Cohen's d 0.42, p=0.048). Compared to type A, type B alexithymia was also associated with a higher prevalence of self-reported diagnosis of major depressive- (30.2% vs. 8.3%) and anxiety disorder (18.9% vs. 3.3%). CONCLUSIONS: The results of this study support the hypothesis of alexithymia subtypes, and add support to the growing evidence showing that alexithymia is likely a heterogeneous and dimensional phenomenon. The subtype (type B) with most pronounced difficulties in identifying feelings may be associated with a higher risk for psychiatric illness compared to type A alexithymia, and may exhibit a more severe history of early life adversity.

Pregnancy-related anxiety and depressive symptoms are associated with visuospatial working memory errors during pregnancy.

BACKGROUND: Cognitive deficits, especially in memory and concentration, are often reported during pregnancy. Similar cognitive dysfunctions can also occur in depression and anxiety. To date, few studies have investigated the associations between cognitive deficits and psychiatric symptoms during pregnancy. This field is of interest because maternal cognitive functioning, and particularly its higher-order aspects are related to maternal well-being and caregiving behavior, as well as later child development. METHODS: Pregnant women (N =230), reporting low (n =87), moderate (n =97), or high (n =46) levels of depressive, general anxiety and/or pregnancy-related anxiety symptoms (assessed repeatedly with EPDS, SCL-90/anxiety subscale, PRAQ-R2, respectively) were tested in mid-pregnancy for their cognitive functions. A computerized neuropsychological test battery was used. RESULTS: Pregnant women with high or moderate level of psychiatric symptoms had significantly more errors in visuospatial working memory/executive functioning task than mothers with low symptom level. Depressive symptoms throughout pregnancy and concurrent pregnancy-related anxiety symptoms were significant predictors of the performance in the task. General anxiety symptoms were not related to visuospatial working memory. LIMITATIONS: Cognitive functions were evaluated only at one time-point during pregnancy precluding causal conclusions. CONCLUSIONS: Maternal depressive symptoms and pregnancy-related anxiety symptoms were both associated with decrements in visuospatial working memory/executive functioning. Depressive symptoms seem to present more stable relationship with cognitive deficits, while pregnancy-related anxiety was associated only concurrently. Future studies could investigate, how stable these cognitive differences are, and whether they affect maternal ability to deal with demands of pregnancy and later parenting.

Non-destructive and direct determination of the degree of substitution of carboxymethyl cellulose by HR-MAS 13C NMR spectroscopy.

We report on the direct assessment of the degree of substitution (DS) of carboxymethyl cellulose (CMC) by High Resolution Magic Angle Spinning (HR-MAS) 13C NMR spectroscopy. The method is applied to industrial CMCs with low and high viscosity and nominal DS, purified and technical samples, and from cellulose linters or wood. The preparation of a set of purified CMC working standards with accurate DS values for the method validation is also described. The DS values determined via HR-MAS 13C NMR on the industrial samples are critically compared to the corresponding values achieved through the USP 37 〈281〉 method (ASH method) and the HPLC method, and the advantages and limitations of the HR-MAS NMR method highlighted. Finally, the HR-MAS NMR approach allowed the accurate DS assessment in CMC with low DS, characterized by a non-negligible fraction of non-functionalized cellulose. The proposed "effective DS" accounts for the DS of the solvent-exposed CMC.

Activation of myeloid and endothelial cells by CD40L gene therapy supports T-cell expansion and migration into the tumor microenvironment.

CD40 is an interesting target in cancer immunotherapy due to its ability to stimulate T-helper 1 immunity via maturation of dendritic cells and to drive M2 to M1 macrophage differentiation. Pancreatic cancer has a high M2 content that has shown responsive to anti-CD40 agonist therapy and CD40 may thus be a suitable target for immune activation in these patients. In this study, a novel oncolytic adenovirus armed with a trimerized membrane-bound extracellular CD40L (TMZ-CD40L) was evaluated as a treatment of pancreatic cancer. Further, the CD40L mechanisms of action were elucidated in cancer models. The results demonstrated that the virus transferring TMZ-CD40L had oncolytic capacity in pancreatic cancer cells and could control tumor progression. TMZ-CD40L was a potent stimulator of human myeloid cells and T-cell responses. Further, CD40L-mediated stimulation increased tumor-infiltrating T cells in vivo, which may be due to a direct activation of endothelial cells to upregulate receptors for lymphocyte attachment and transmigration. In conclusion, CD40L-mediated gene therapy is an interesting concept for the treatment of tumors with high levels of M2 macrophages, such as pancreatic cancer, and an oncolytic virus as carrier of CD40L may further boost tumor killing and immune activation.

Altered heart proteome in fructose-fed Fisher 344 rats exposed to bisphenol A.

Bisphenol A (BPA), is an artificial estrogen initially produced for medical purposes but is today widely used in polycarbonate plastics and epoxy resins. Exposure-related reproductive disorders have been found, but recently it has also been suggested that BPA may be involved in obesity, diabetes, myocardial hypertrophy and myocardial infarction in humans. To mimic a modern lifestyle, female rats were fed with fructose or fructose plus BPA (0.25mg/L drinking water). The myocardial left ventricle proteome of water controls, fructose-fed and fructose-fed plus BPA supplemented rats was explored. The proteome was investigated using nano-liquid chromatography tandem mass spectrometry and two-dimensional gel electrophoresis followed by matrix assisted laser desorption/ionization mass spectrometry identification. In total, 41 proteins were significantly altered by BPA exposure compared to water or fructose controls. Principal component analysis and cellular process enrichment analysis of altered proteins suggested increased fatty acid transport and oxidation, increased ROS generation and altered structural integrity of the myocardial left ventricle in the fructose-fed BPA-exposed rats, indicating unfavorable effects on the myocardium. In conclusion, BPA exposure in the rats induces major alterations in the myocardial proteome.

Adaption of pregnancy anxiety questionnaire-revised for all pregnant women regardless of parity: PRAQ-R2.

The 10-item Pregnancy-Related Anxiety Questionnaire-Revised (PRAQ-R) is a widely used instrument to assess and identify pregnancy-specific anxiety in nulliparous women. It has good psychometric values and predictive validity for birth and childhood outcomes. Nonetheless, the PRAQ-R is not designed for use in parous women, as particularly one item of the questionnaire is not relevant for women who gave birth before. We tested the factorial and scalar invariance of a modified PRAQ-R2 across nulliparous and parous women with an adapted item to fit both groups of pregnant women. A longitudinal study among 1144 pregnant women (n = 608 nulliparous and n = 536 parous) with two repeated measures of the PRAQ-R2 was used to test for measurement invariance of the instrument. Results show metric and scalar invariance, indicating that the PRAQ-R2 measures similar constructs on the same scale for all pregnant women at two different times during pregnancy. We conclude that the PRAQ-R2 can be used, compared, or combined in a sample of nulliparous and parous women.

Weight loss after bariatric surgery normalizes brain opioid receptors in morbid obesity.

Positron emission tomography (PET) studies suggest opioidergic system dysfunction in morbid obesity, while evidence for the role of the dopaminergic system is less consistent. Whether opioid dysfunction represents a state or trait in obesity remains unresolved, but could be assessed in obese subjects undergoing weight loss. Here we measured brain µ-opioid receptor (MOR) and dopamine D2 receptor (D2R) availability in 16 morbidly obese women twice-before and 6 months after bariatric surgery-using PET with [(11)C]carfentanil and [(11)C]raclopride. Data were compared with those from 14 lean control subjects. Receptor-binding potentials (BPND) were compared between the groups and between the pre- and postoperative scans among the obese subjects. Brain MOR availability was initially lower among obese subjects, but weight loss (mean=26.1 kg, s.d.=7.6 kg) reversed this and resulted in ~23% higher MOR availability in the postoperative versus preoperative scan. Changes were observed in areas implicated in reward processing, including ventral striatum, insula, amygdala and thalamus (P's<0.005). Weight loss did not influence D2R availability in any brain region. Taken together, the endogenous opioid system plays an important role in the pathophysiology of human obesity. Because bariatric surgery and concomitant weight loss recover downregulated MOR availability, lowered MOR availability is associated with an obese phenotype and may mediate excessive energy uptake. Our results highlight that understanding the opioidergic contribution to overeating is critical for developing new treatments for obesity.

In vitro discovery of promising anti-cancer drug combinations using iterative maximisation of a therapeutic index.

In vitro-based search for promising anti-cancer drug combinations may provide important leads to improved cancer therapies. Currently there are no integrated computational-experimental methods specifically designed to search for combinations, maximizing a predefined therapeutic index (TI) defined in terms of appropriate model systems. Here, such a pipeline is presented allowing the search for optimal combinations among an arbitrary number of drugs while also taking experimental variability into account. The TI optimized is the cytotoxicity difference (in vitro) between a target model and an adverse side effect model. Focusing on colorectal carcinoma (CRC), the pipeline provided several combinations that are effective in six different CRC models with limited cytotoxicity in normal cell models. Herein we describe the identification of the combination (Trichostatin A, Afungin, 17-AAG) and present results from subsequent characterisations, including efficacy in primary cultures of tumour cells from CRC patients. We hypothesize that its effect derives from potentiation of the proteotoxic action of 17-AAG by Trichostatin A and Afungin. The discovered drug combinations against CRC are significant findings themselves and also indicate that the proposed strategy has great potential for suggesting drug combination treatments suitable for other cancer types as well as for other complex diseases.