Impacts of metal mining on river systems: a global assessment.

An estimated 23 million people live on floodplains affected by potentially dangerous concentrations of toxic waste derived from past and present metal mining activity. We analyzed the global dimensions of this hazard, particularly in regard to lead, zinc, copper, and arsenic, using a georeferenced global database detailing all known metal mining sites and intact and failed tailings storage facilities. We then used process-based and empirically tested modeling to produce a global assessment of metal mining contamination in river systems and the numbers of human populations and livestock exposed. Worldwide, metal mines affect 479,200 kilometers of river channels and 164,000 square kilometers of floodplains. The number of people exposed to contamination sourced from long-term discharge of mining waste into rivers is almost 50 times greater than the number directly affected by tailings dam failures.

Outcomes and prognostic factors in patients with haematological malignancy admitted to a specialist cancer intensive care unit: a 5 yr study.

BACKGROUND: Long-held assumptions of poor prognoses for patients with haematological malignancies (HM) have meant that clinicians have been reluctant to admit them to the intensive care unit (ICU). We aimed to evaluate ICU, in-hospital, and 6 month mortality and to identify predictors for in-hospital mortality. METHODS: A cohort study in a specialist cancer ICU of adult HM patients admitted over 5 yr. Data acquired included: patient characteristics, haematological diagnosis, haematopoietic stem cell transplant (HSCT), reason for ICU admission, and APACHE II scores. Laboratory values, organ failures, and level of organ support were recorded on ICU admission. Predictors for in-hospital mortality were evaluated using uni- and multivariate analysis. RESULTS: Of 199 patients, median age was 58 yr [inter-quartile range (IQR) 46-66], 51.7% were emergency admissions, 42.2% post-HSCT, 51.9% required mechanical ventilation, median APACHE II was 21 (IQR 16-25), and median organ failure numbered 2 (IQR 1-4). ICU, in-hospital, and 6 month mortalities were 33.7%, 45.7%, and 59.3%, respectively. Univariate analysis revealed bilirubin >32 µmol litre(-1), mechanical ventilation, ≥2 organ failures, renal replacement therapy, vasopressor support (all P<0.001), graft-vs-host disease (P=0.007), APACHE II score (P=0.02), platelets ≤20×10(9) litre(-1) (P=0.03), and proven invasive fungal infection (P=0.04) were associated with in-hospital mortality. Multivariate analysis revealed that ≥2 organ failures [odds ratio (OR) 5.62; 95% confidence interval (95% CI), 2.30-13.70] and mechanical ventilation (OR 3.03; 95% CI, 1.33-6.90) were independently associated with in-hospital mortality. CONCLUSIONS: Mortality was lower than in previous studies. Mechanical ventilation and ≥2 organ failures were independently associated with in-hospital mortality. 'Traditional' variables such as neutropenia, transplantation status, and APACHE II score no longer appear to be predictive.

The effect of explantation on systemic disease symptoms and quality of life in patients with breast implant illness: a prospective cohort study.

Silicone breast implants (SBIs) have been subject to scientific scrutiny since the 1960's because of their potential link with systemic disease symptoms. Breast implant illness (BII) is a cluster of over 56 (systemic) symptoms attributed by patients to their SBIs. BII remains an unofficial medical diagnosis, although its symptoms include but are not limited to the clinical manifestations of autoimmune/inflammatory syndrome induced by adjuvants (ASIA). The aim of this study was to prospectively analyse the effect of explantation on clinical manifestations of ASIA/BII symptoms, as well as to compare (breast-surgery specific) QoL in patients pre- and postoperatively while recording relevant perioperative/patient data. A prospective cohort study was conducted on 140 patients consulting a single surgeon for explantation of SBIs at a single clinic from 2019 to 2021 via their general practitioner, a medical specialist or self-referral. Of all patients, medical (implant) history, lifestyle factors and biometric data were obtained. Patients filled out a novel ASIA/BII symptom-survey termed the ASIA-scale, three domains of the SF-36 and the augmentation module of the BREAST-Q before and four months after the operation. A total of 109 patients completed both the pre- and postoperative survey with a mean follow-up duration of 205 days. There was a significant decrease in all individual symptom scores as well as ASIA-scale summary scores after explantation (p < .001). All SF-36 subdomains showed significant improvement postoperatively (p < .001). The BREAST-Q subdomain 'satisfaction with breasts' improved significantly after explantation (p = .036). No statistically significant association was found between any clinical parameters (such as age, capsulectomy, rupture etc.) and the recovery of symptom scores. This is the largest prospective cohort study on SBI explantation to date showing significant improvement of the most common systemic complaints in SBI patients as well as improvement of satisfaction with breasts and overall quality of life.

Measuring interoception: The phase adjustment task.

Interoception, perception of one's bodily state, has been associated with mental health and socio-emotional processes. However, several interoception tasks are of questionable validity, meaning associations between interoception and other variables require confirmation with new measures. Here we describe the novel, smartphone-based Phase Adjustment Task (PAT). Tones are presented at the participant's heart rate, but out of phase with heartbeats. Participants adjust the phase relationship between tones and heartbeats until they are synchronous. Data from 124 participants indicates variance in performance across participants which is not affected by physiological or strategic confounds. Associations between interoception and anxiety, depression and stress were not significant. Weak associations between interoception and mental health variables may be a consequence of testing a non-clinical sample. A second study revealed PAT performance to be moderately stable over one week, consistent with state effects on interoception.

Mechanistic validation of a clinical lead stapled peptide that reactivates p53 by dual HDM2 and HDMX targeting.

Hydrocarbon-stapled peptides that display key residues of the p53 transactivation domain have emerged as bona fide clinical candidates for reactivating the tumor suppression function of p53 in cancer by dual targeting of the negative regulators HDM2 and HDMX. A recent study questioned the mechanistic specificity of such stapled peptides based on interrogating their capacity to disrupt p53/HDM2 and p53/HDMX complexes in living cells using a new recombinase enhanced bimolecular luciferase complementation platform (ReBiL). Here, we directly evaluate the cellular uptake, intracellular targeting selectivity and p53-dependent cytotoxicity of the clinical prototype ATSP-7041. We find that under standard serum-containing tissue culture conditions, ATSP-7041 achieves intracellular access without membrane disruption, dose-dependently dissociates both p53/HDM2 and p53/HDMX complexes but not an unrelated protein complex in long-term ReBiL experiments, and is selectively cytotoxic to cancer cells bearing wild-type p53 by inducing a surge in p53 protein level. These studies underscore the importance of a thorough stepwise approach, including consideration of the time-dependence of cellular uptake and intracellular distribution, in evaluating and advancing stapled peptides for clinical translation.

Molecular chirality and charge transfer through self-assembled scaffold monolayers.

The effect of molecular chirality on electron transmission is explored by photoelectrochemistry. Thiol-terminated chiral scaffold molecules containing a porphyrin chromophore were self-assembled on gold surfaces to form a monolayer. Incorporation of the SAM-coated gold into an electrochemical cell and illumination with visible light generated a cathodic photocurrent. When using circularly polarized light, the photocurrent displayed an asymmetry (different magnitude of photocurrent for right versus left polarization) that changed with the molecular chirality (left- or right-handedness of the scaffold). A symmetry constraint on the electronic coupling between the porphyrin and the organic scaffold is proposed as a possible mechanism for the photocurrent asymmetry.

Somatosensory activations during the observation of touch and a case of vision-touch synaesthesia.

In this study, we describe a new form of synaesthesia in which visual perception of touch elicits conscious tactile experiences in the perceiver. We describe a female subject (C) for whom the observation of another person being touched is experienced as tactile stimulation on the equivalent part of C's own body. Apart from this clearly abnormal synesthetic experience, C is healthy and normal in every other way. In this study, we investigate whether C's 'mirrored touch' synesthetic experience is caused by overactivity in the neural system that responds to the observation of touch. A functional MRI experiment was designed to investigate the neural system involved in the perception of touch in a group of 12 non-synesthetic control subjects and in C. We investigated neural activity to the observation of touch to a human face or neck compared with the observation of touch to equivalent regions on an object. Furthermore, to investigate the somatosensory topography of the activations during observation of touch, we compared activations when observing a human face or neck being touched with activations when the subjects themselves were touched on their own face or neck. The results demonstrated that the somatosensory cortex was activated in the non-synesthetic subjects by the mere observation of touch and that this activation was somatotopically organized such that observation of touch to the face activated the head area of primary somatosensory cortex, whereas observation of touch to the neck did not. Moreover, in non-synesthetic subjects, the brain's mirror system-comprising premotor cortex, superior temporal sulcus and parietal cortex-was activated by the observation of touch to another human more than to an object. C's activation patterns differed in three ways from those of the non-synesthetic controls. First, activations in the somatosensory cortex were significantly higher in C when she observed touch. Secondly, an area in left premotor cortex was activated in C to a greater extent than in the non-synesthetic group. Thirdly, the anterior insula cortex bilaterally was activated in C, but there was no evidence of such activation in the non-synesthetic group. The results suggest that, in C, the mirror system for touch is overactive, above the threshold for conscious tactile perception.

Acquired alexithymia following damage to the anterior insula.

Alexithymia is a subclinical condition characterized by impaired awareness of one's emotional states, which has profound effects on mental health and social interaction. Despite the clinical significance of this condition, the neurocognitive impairment(s) that lead to alexithymia remain unclear. Recent theoretical models suggest that impaired anterior insula (AI) functioning might be involved in alexithymia, but conclusive evidence for this hypothesis is lacking. We measured alexithymia levels in a large sample of brain-injured patients (N=129) and non-brain-injured control participants (N=33), to determine whether alexithymia can be acquired after pronounced damage to the AI. Alexithymia levels were first analysed as a function of group, with patients separated into four groups based on AI damage: patients with >15% damage to AI, patients with <15% damage to AI, patients with no damage to AI, and healthy controls. An ANOVA revealed that alexithymia levels varied across groups (p=0.009), with >15% AI damage causing higher alexithymia relative to all other groups (all p<0.01). Next, a multiple linear regression model was fit with the degree of damage to AI, the degree of damage to a related region (the anterior cingulate cortex, ACC), and the degree of damage to the whole brain as predictor variables, and alexithymia as the dependent variable. Critically, increased AI damage predicted increased alexithymia after controlling for the other two regressors (ACC damage; total lesion volume). Collectively, our results suggest that pronounced AI damage causes increased levels of alexithymia, providing critical evidence that this region supports emotional awareness.

XactMice: humanizing mouse bone marrow enables microenvironment reconstitution in a patient-derived xenograft model of head and neck cancer.

The limitations of cancer cell lines have led to the development of direct patient-derived xenograft models. However, the interplay between the implanted human cancer cells and recruited mouse stromal and immune cells alters the tumor microenvironment and limits the value of these models. To overcome these constraints, we have developed a technique to expand human hematopoietic stem and progenitor cells (HSPCs) and use them to reconstitute the radiation-depleted bone marrow of a NOD/SCID/IL2rg(-/-) (NSG) mouse on which a patient's tumor is then transplanted (XactMice). The human HSPCs produce immune cells that home into the tumor and help replicate its natural microenvironment. Despite previous passage on nude mice, the expression of epithelial, stromal and immune genes in XactMice tumors aligns more closely to that of the patient tumor than to those grown in non-humanized mice-an effect partially facilitated by human cytokines expressed by both the HSPC progeny and the tumor cells. The human immune and stromal cells produced in the XactMice can help recapitulate the microenvironment of an implanted xenograft, reverse the initial genetic drift seen after passage on non-humanized mice and provide a more accurate tumor model to guide patient treatment.

Calcium mobilization by inositol phosphates and other intracellular messengers.

Inositol 1,4,5-trisphosphate [Ins(1,4,5)P(3)] is now widely recognized as a messenger controlling the release of calcium from intracellular stores. In oocytes, and also probably in excitable cells, another potential calcium-mobilizing messenger is cyclic ADP ribose, although there is as yet little evidence that its levels are regulated by hormones or other extracellular mediators. In addition to signaling intracellular calcium release, [Ins(1,4,5)P(3)] also regulates calcium entry across the plasma membrane, but not in a direct manner. Rather, the depletion of intracellular stores by the calcium-mobilizing action of [Ins (1,4,5)P(3)] initiates a process of retrograde signaling whereby the depleted stores generate or release a diffusible messenger that is believed to act on the plasma membrane. A phosphorylated metabolite of [Ins(1,4,5)P(3)], inositol 1,3,4,5-tetrakisphosphate [Ins(1,3,4,5)P(4)], has been proposed to modulate this process, but the literature is not consistent on this point. A recently proposed candidate for the retrograde messenger is an activity extracted from Jurkat cells termed CIF (calcium influx factor), which has many properties consistent with such a messenger. There is also evidence that a GTP-dependent process, possibly involving a small G protein, is involved in signaling calcium entry and may be involved in either the formation or action of the diffusible messenger for calcium entry.

cGMP is not required for capacitative Ca2+ entry in Jurkat T-lymphocytes.

In this study, we have addressed the potential role of cGMP in regulating calcium entry in Jurkat T-lymphocytes. These cells display capacitative Ca(2+)-entry in response to the intracellular Ca(2+)-ATPase inhibitor, thapsigargin (TG). In the presence of extracellular Ca2+, TG stimulates a sustained elevation of intracellular cGMP levels. In the absence of extracellular Ca2+, TG induces no apparent increase in the levels of cGMP. However, experiments using Mn2+, as a surrogate for Ca2+, demonstrated that TG increased the rate of divalent cation entry in the absence of extracellular Ca2+. Treatment of Jurkat cells with the guanylyl cyclase inhibitor, LY83583 (20 microM), completely blocked cGMP formation in response to TG. However, LY83583 treated cells still exhibited a sustained, albeit partially reduced, Ca2+ response induced by TG. These data demonstrate that, in Jurkat cells, the sustained formation of cGMP is dependent on elevated intracellular Ca2+, and that elevated levels of cGMP are not necessary for the activation of capacitative Ca2+ entry.

Effect of cytoplasmic Ca2+ on (1,4,5)IP3 formation in vasopressin-activated hepatocytes.

The role of cytoplasmic calcium as a regulator of phospholipase C in vasopressin-activated hepatocytes was examined. According to models in which calcium spiking arises because of a positive feedback by calcium on phospholipase C, Ca2+ is seen as a positive modulator of phospholipase C under conditions of submaximal receptor activation. However, in hepatocytes whose precursor lipids had been labeled by incubation in [3H]-inositol, no increase in [3H]-(1,4,5)IP3 was detected in response to thapsigargin, in either unstimulated cells, or in cells stimulated with 1 nM vasopressin. Addition of a maximal concentration of vasopressin (1 microM) caused a rapid and substantial increase in [3H]-(1,4,5)IP3. These results indicate that changes in cytoplasmic calcium do not influence phospholipase C activity in hepatocytes, even under conditions of submaximal agonist activation. These findings also support models that provide for calcium spiking at constant levels of (1,4,5)IP3 at least in the case of the rat hepatocyte.

Mechanisms of activated Ca2+ entry in the rat pancreatoma cell line, AR4-2J.

The characteristics of Ca2+ entry activated by surface receptor agonists and membrane depolarization were studied in the rat pancreatoma cell line, AR4-2J. Ca2+ mobilization activated by substance P, bombesin, or muscarinic receptor stimulation was found to involve both Ca2+ release and entry. In addition, depolarization of the surface membrane of AR4-2J cells with elevated concentrations of K+ activated Ca2+ entry. Ca2+ entry induced by membrane depolarization was inhibited by the L-channel antagonist, nimodipine, while that due to surface receptor agonists was not inhibited by this agent. The microsomal Ca(2+)-ATPase inhibitor, thapsigargin, caused both depletion of the agonist-sensitive intracellular Ca2+ pool and sustained Ca2+ influx indistinguishable from that produced by bombesin or methacholine. These results confirm that, unlike the pancreatic acinar cells from which they are presumably derived, AR4-2J cells express voltage-sensitive, dihydropyridine-inhibitable Ca2+ channels. However, in contrast to previous reports with this cell line, in the AR4-2J cells in use in our laboratory, and under our experimental conditions, surface receptor agonists (including substance P) do not cause Ca2+ influx through voltage-sensitive Ca2+ channels. Instead, we conclude that agonist-activated Ca2+ mobilization is initiated by (1,4,5)IP3-mediated intracellular Ca2+ release and that Ca2+ influx is regulated primarily, if not exclusively, by the state of depletion of the (1,4,5)IP3-sensitive intracellular Ca2+ pool.

Effects of elevated cytoplasmic calcium and protein kinase C on endoplasmic reticulum structure and function in HEK293 cells.

In human embryonic kidney (HEK) cells stably transfected with green fluorescent protein targeted to the endoplasmic reticulum (ER), elevation of intracellular Ca2+ ([Ca2+]i) altered ER morphology, making it appear punctate. Electron microscopy revealed that these punctate structures represented circular and branched rearrangements of the endoplasmic reticulum, but did not involve obvious swelling or pathological fragmentation. Activation of protein kinase C with phorbol 12-myristate 13-acetate (PMA), prevented the effects of ionomycin on ER structure without affecting the elevation of [Ca2+]i. These results suggest that protein kinase C activation alters cytoplasmic or ER components underlying the effects of high [Ca2+]i on ER structure. Treatment of HEK cells with PMA also reduced the size of the thapsigargin-sensitive Ca2+ pool and inhibited Ca2+ entry in response to thapsigargin. Thus, protein kinase C activation has multiple actions on the calcium storage and signalling function of the endoplasmic reticulum in HEK cells: (1) reduced intracellular Ca2+ storage capacity, (2) inhibition of capacitative Ca2+ entry, and (3) protection of the endoplasmic reticulum against the effects of high [Ca2+]i.

D2 dopamine receptor gene TaqI A1 and B1 restriction fragment length polymorphisms: enhanced frequencies in psychostimulant-preferring polysubstance abusers.

Several lines of evidence suggest that presence of a D2 dopamine receptor (DRD2) gene variant marked by TaqI restriction fragment length polymorphisms (RFLPs) might contribute to vulnerability to substance abuse. Psychostimulants display the most robust enhancement of dopamine activity in mesolimbic/mesocortical circuits important for behavioral reward. The present study tests the hypothesis that a DRD2 gene variant might be more prominent in polysubstance users who preferentially use psychostimulants than in addicts with preferential opiate use or in those with no drug preference. Polysubstance users with histories of heavy daily preferential psychostimulant use more often displayed one or two copies of the TaqI A1 (27/62 = 43.5% vs 33/119 = 27.7% for controls), and B1 (20/62 = 32.3% vs 23/119 = 19.8% for controls) markers at the DRD2 locus. DRD2 gene marker distributions in abusers with more prominent opiate use, or those with no history of drug preference, were similar to control genotypes. Psychostimulant-preferring drug users also reported earlier onset of psychostimulant use. Our data are consistent with the hypothesis that DRD2 gene variants marked by these polymorphisms may work, probably in concert with other genetic and environmental factors, to enhance vulnerability to psychostimulant abuse.

A human synaptic vesicle monoamine transporter cDNA predicts posttranslational modifications, reveals chromosome 10 gene localization and identifies TaqI RFLPs.

A human vesicular monoamine transporter cDNA has been identified by screening a human brainstem library using sequences from the rat brain synaptic vesicle monoamine transporter (SVMT) [(1992) Cell 70, 539-551; (1992) Proc. Natl. Acad. Sci. USA 89, 10993-10997]. The hSVMT shares 92% amino acid identity with the rat sequence, but displays one less consensus site for asparagine N-linked glycosylation and one more consensus site for phosphorylation by protein kinase C. The human SVMT gene maps to chromosome 10q25 using Southern blotting analysis of human/rodent hybrid cell lines and fluorescent in situ hybridization approaches. The cDNA, and a subclone, recognize TaqI polymorphisms that may prove useful to assess this gene's involvement in neuropsychiatric disorders involving monoaminergic brain systems.

Modulation of primary afferent-mediated neurotransmission and Fos expression by glutamate uptake inhibition in rat spinal neurones in vitro.

The effect of altered endogenous levels of synaptic glutamate on neurotransmission and synaptic dorsal horn Fos expression was determined in rat spinal cord in vitro. The uptake inhibitor L-trans-pyrrolidine-2,4-dicarboxylate (L-PDC, 1mM) was tested against dorsal root-ventral root potentials (DR-VRP), afferent-mediated slow dorsal horn excitatory postsynaptic potentials (DR-EPSP) and nociceptive afferent-induced synaptic currents (EPSCs) of substantia gelatinosa neurones. L-PDC reduced DR-VRP fast and slow peak amplitude and duration (P<0.05), slow DR-EPSP amplitude and duration (P<0.005) and EPSC amplitude (P<0.05). The Group II/III mGluR antagonist (RS)-alpha-cyclopropyl-4-phosphonophenylglycine (CPPG, 100 microM) reduced L-PDC inhibition of synaptic potentials. The Group II antagonist (2S)-2-amino-2-(1S,2S-2-carboxycycloprop-1-yl)-3-(xanth-9-yl)propanoic acid (LY341495, 300 nM) and the Group III antagonist (RS)-alpha-methylserine-O-phosphate (MSOP, 10 microM) partially reversed EPSC inhibition by L-PDC. The Group III agonist L(+)-2-amino-4-phosphonobutyric acid (L-AP4, 30 microM) mimicked CPPG-sensitive inhibitory effects of L-PDC on DR-VRP (P<0.001) and the slow DR-EPSP (P<0.005). L-PDC (1mM) or L-AP4 (30 microM) reduced afferent-evoked dorsal horn Fos expression, this effect was reversed by CPPG. These data suggest that increased synaptic glutamate levels may activate inhibitory Group II/III mGluR receptors and impact significantly on nociceptive neurotransmission and transcriptional adaptive responses of target neurones.

Coherent backscatter enhances reflection confocal microscopy.

When thin optically transparent specimens are grown on reflective substrates, contrast in reflection confocal microscopy is markedly enhanced. This enhanced contrast allows for the visualization of the thin filopodia and organelles contained within the neuritic processes of PC12 cells in culture. The characteristics of this contrast enhancement suggest that it arises because of interference between light scattered from the specimen and coherently backscattered illumination reflected off the substrate. This technique provides a method for visualizing living cells or other similarly transparent objects on opaque substrates in a nondestructive manner.

Human and mouse dopamine transporter genes: conservation of 5'-flanking sequence elements and gene structures.

Synaptic reaccumulation of the neurotransmitter dopamine is mediated by the dopamine transporter (DAT), a member of the family of twelve transmembrane domain, sodium- and chloride-dependent neurotransmitter transporters. Several DAT features, including its exclusive expression in dopaminergic neurons, implication in cocaine action, and prominent role in the mechanisms of Parkinsonism-inducing neurotoxins, make understanding of the DAT gene of interest. Isolation and characterization of the human and mouse DAT genes has allowed elucidation of similarities between each and other members of this transporter gene family. Sequences 5' to transcriptional start sites contain G-C rich, TATA-less, CAAT-less regions with striking conservation between human and mouse gene flanking regions. These studies suggest sequence elements that are candidates to contribute to the dopamine transporter's dopaminergic cell-specific expression.