A curious association of chronic homeopathic arsenic ingestion with nonspecific symptoms in a Swiss teenager.

Arsenic is a toxicant that has no dose threshold below which exposures are not harmful. Here I report a curious association of chronic homeopathic arsenic ingestion with nonspecific symptoms in a Swiss teenager. For about 4 years she had taken globules of a freely purchasable homeopathic remedy containing inorganic arsenic (iAs), infinitesimally diluted to D6 (average arsenic content per single globule: 0.85 ± 0.08 ng). In the previous 7 months she had taken 20 to 50 globules daily (average 30 ng arsenic daily). She complained of nausea, stomach and abdominal cramps, diarrhoea and flatulence, headache, dizziness, anxiety, difficulty concentrating, insomnia, snoring, leg cramps and fatigue, loss of appetite, increased thirst and sweating, reduced diuresis, weight gain, paleness and coolness of both hands with a furry feeling of the hands, eczema of the hands, arms and legs, conjunctivitis and irregular menstruation. The physical and laboratory examinations showed a body mass index of 30 kg/m2, acne vulgaris, bilateral spotted leukonychia, eczema of hands, arms and legs, non-pitting oedema of the legs, elevated plasma alkaline phosphatase activity, folate deficiency and severe vitamin D3 insufficiency. The arsenic concentration in her blood was <0.013 µmol/l, and arsenic was undetectable in her scalp hair. The total iAs concentration was 116 nmol/l in the morning urine and 47 nmol/l in the afternoon urine. The urinary arsenic concentration decreased and the patient’s complaints improved upon interruption of the arsenic globules, vitamin D3, thiamine and folic acid supplementation, and symptomatic therapy. It is concluded that an avoidable toxicant such as inorganic arsenic, for which no scientific safe dose threshold exists, should be avoided and not be found in over-the-counter medications.

The arsenic for phosphorus swap is accidental, rather than a facultative one, and the question whether arsenic is nonessential or toxic is quantitative, not a qualitative one.

Arsenic shares many physicochemical properties with phosphorus, so that arsenic can be taken up inadvertently by cells through the pathways for phosphorus. As a phosphate analog, arsenate competes with phosphate and enters cells via phosphate transporters. In the cell, arsenate can be recognized as a substrate by enzymes that usually use phosphate as a substrate. The phosphate for arsenate swap results in wasteful 'futile cycles' in metabolic pathways, uncoupled oxidative phosphorylation and extreme DNA instability. The disrupting metabolic effects of arsenic have an evolutionary meaning, so that all living organisms-from chemoautotrophic organisms that grow by reducing or oxidizing arsenic to metazoan--carry highly conserved arsenic resistance genes. Arsenic resistance can result from different strategies including selective transport to maximize phosphate uptake and minimize entry of arsenate, active transport to export arsenate, arsenic storage in specialized compartments, enzyme selectivity toward phosphate, and increased efficiency of DNA repair systems. None of these strategies is infallible, though, and susceptibility to arsenic toxicity varies between taxa in many orders of magnitude. Even arsenic-hypertolerant organisms will stop to grow and will eventually die when exposed to arsenic over species-specific resistance limits. The arsenic for phosphorus swap is an accidental one, it does not warrant a conclusion in favor of the essentiality of arsenic to life as we know it.

Arsenic for the fool: an exponential connection.

Anthropogenic arsenic is insidiously building up together with natural arsenic to a level unprecedented in the history of mankind. Arsenopyrite (FeAsS) is the principal ore of arsenic and gold in hard rock mines; it is formed by a coupled substitution of sulphur by arsenic in the structure of pyrite (FeS(2)) - nicknamed "fool's gold". Other important sources of anthropogenic arsenic are fossil fuels such as coal and oil. Here I report on the first indication that the environmental concentration of total arsenic in topsoils - in the 7-18ppm range - is exponentially related to the prevalence and mortality of Alzheimer's disease and other dementias in European countries. This evidence defies the imputed absence of verified cases of human morbidity or mortality resulting from exposure to low-level arsenic in topsoils.

Gold, coal and oil.

Jared Diamond has hypothesized that guns, germs and steel account for the fate of human societies. Here I propose an extension of Diamond's hypothesis and put it in other terms and dimensions: gold, coal and oil account not only for the fate of human societies but also for the fate of mankind through the bodily accumulation of anthropogenic arsenic, an invisible weapon of mass extinction and evolutionary change. The background is clear; arsenic species fulfill seven criteria for a weapon of mass extinction and evolutionary change: (i) bioavailability to all living organisms; (ii) imperceptibility; (iii) acute toxicity; (iv) bioaccumulation and chronic toxicity; (v) adverse impact on reproductive fitness and reproductive outcomes and early-age development and growth in a wide range of microbial, plant and animal species including man; (vi) widespread geographical distribution, mobility and ecological persistence on a centennial to millennial basis and (vii) availability in necessary and sufficient amounts to exert evolutionarily meaningful effects. The proof is becoming increasingly feasible as human exploitation of gold, coal and oil deposits cause sustainable rises of arsenic concentrations in the biosphere. Paradoxically, humans are among the least arsenic-resistant organisms because humans are long-lived, encephalized and complex social metazoans. An arsenic accumulation model is presented here to describe how arsenic accumulates in the human body with increasing age and at different provisionally safe exposure levels. Arsenic accumulates in the human body even at daily exposure levels which are within the lowest possible WHO provisional tolerance limits, yielding bodily arsenic concentrations which are above WHO provisional limits. Ongoing consequences of global scale arsenic poisoning of mankind include age-specific rises in morbidity and mortality followed by adaptive changes. The potential rise of successful forms of inborn resistance to arsenic in humans will make it certain that a number of other hardly won, nicely balanced human-specific adaptednesses will decline. These include a decline of encephalization and life-span, and consequentially intelligence and longevity. These changes are likely to have far-reaching impacts on biological and cultural evolution of mankind. The only efficient way of reducing chronic global exposure to arsenic and avoiding further human losses is the inactivation of important sources of anthropogenic arsenic such as hard rock mining and burning of fossil fuels.

Less DeltamtDNA4977 than normal in various types of tumors suggests that cancer cells are essentially free of this mutation.

Levels of mtDNA(4977) deletions (DeltamtDNA(4977)) have been found to be lower in tumors than in adjacent non-tumoral tissues. In 87 cancer patients, DeltamtDNA(4977) was detected by multiplex polymerase chain reaction (PCR) amplification in 43 (49%) of the tumors and in 74 (85%) of the samples of non-tumoral tissues that were adjacent to the tumors. DeltamtDNA(4977) deletions were detected in 24% of the breast tumors, 52% of the colorectal tumors, 79% of the gastric tumors, and 40% of the head and neck tumors as compared with 77, 83, 100, and 90% of the adjacent respective non-tumoral tissues at the same DNA template dilution. Based on limiting dilution PCR of 16 tumors and their adjacent non-tumoral tissues, it was found that the amount of DeltamtDNA(4977) was 10- to 100-fold lower in the tumor than in the respective control non-tumoral tissues. Real-time PCR experiments were performed to quantify the number of DeltamtDNA(4977) deletions per cell, by determining the mitochondrial-to-nuclear DNA ratio. In all of the cases of breast, colorectal, gastric, and head and neck cancer the proportion of DeltamtDNA(4977) in tumors was lower than that of the respective non-tumoral tissue. Traces of DeltamtDNA(4977) in tumors were apparently due to contamination of tumor tissue with surrounding non-tumoral tissue, as evidenced by tumor microdissection and in situ PCR techniques, suggesting that tumors are essentially free of this mutation. Although the metabolic effect of DeltamtDNA(4977) may be minimal in normal (non-tumor) tissue, in tissue under stress, such as in tumors, even low levels of DeltamtDNA(4977) deletions may be intolerable.

The common mitochondrial DNA deletion deltamtDNA(4977): shedding new light to the concept of a tumor suppressor mutation.

We propose that the age-related accumulation of deltamtDNA(4977) mutations may serve a protective function against tumor-promoting effects of other somatic mutations. The evidence discussed here is consistent with the concept that deltamtDNA(4977) plays a tumor-suppressor role, thus shedding new light to the concept of a tumor suppressor mutation. This concept may help understand how a tumor-promoting mutation may be able to cause malignant transformation in cells lacking a tumor-suppressor mutation, while the same tumor-promoting mutation can be present in cells that carry a tumor-suppressor mutation, without causing cancer.

Evaluation of a method for high yield purification of largely intact mitochondrial DNA from human placentae.

We developed, and quantitatively and qualitatively evaluated an easily reproducible method for high yield purification of mitochondrial DNA (mtDNA) from human placentae by mechanical tissue disruption, differential centrifugation of mitochondria, enzymatic digestion, phenol extraction and ethanol precipitation. Average mtDNA yields were 2.5 microg/g tissue (without an RNAse treatment step) and 1.5 microg/g tissue (with an RNAse treatment step). This mtDNA migrated as a 16.5-kb isolated band in agarose gels; it yielded fragments of expected sizes after digestion with restriction enzymes; it successfully served as a template in long PCR for amplification of mtDNA sequences, and hybridized to an mtDNA probe in a predictable fashion. MtDNA yields of this method were 10-fold higher than those of previously reported ones for mtDNA purification from freshly obtained human cells and tissues, with the advantage that more placental tissue can be obtained for mtDNA purification than other types of tissue, at lower cost, and with minimal or no ethical issues.

A model system for testing gene vectors using murine tumor cells on the chorioallantoic membrane of the chick embryo.

We developed a model system for testing gene vectors, based on the growth of murine tumors on the chorioallantoic membrane (CAM) of embryonic chickens. The ability of selected murine cells to grow on the CAM was rated according to the following criteria: i) formation of tumor masses; ii) metastasis formation; iii) reproducibility; iv) yield, indicated as the number of embryos surviving to assessment time with visible tumors on the CAM; v) maintainability of the cell, both in the original host and the embryonic chick, or 'shuttle maintainability'; vi) detection by the naked eye, and vii) cost/benefit relation. The murine melanoma cell lineage, B16F10, which efficiently forms distinct, pigmented tumor masses and metastases on the CAM, performed better in this model than the murine B61 cell line. In vitro transduction of B16F10 cells with a recombinant adenovirus carrying a construct of the E. coli LacZ gene followed by inoculation onto the CAM resulted in beta-galactosidase expression in the tumor mass growing on the CAM. This model is potentially applicable to preclinical evaluation of gene vectors, especially for gene therapy of cancer.

Analise de mutações somáticas dos genes APP, presenilina 1 e 2, em tecido encefálico na doença de Alzheimer / Somatic mutationals analysis of genes APP, presenilin 1 and 2, in encephalic tissue in Alzheimer's disease

As bases moleculares da forma esporadica da doenca de Alzheimer (DA) permanecem ainda desconhecidas. Nos formulamos a hipotese de que, em alguns casos esporadicos de DA, uma mutacao somatica nos genes da proteina precursora amiloide (APP), da presenilina 1 (PS-1) e 2 (STM2) (genes envolvidos na DA forma familiar) em uma celula embrionaria comprometida com o desenvolvimento neuronal, pode resultar em fenotipo da DA. Usando a tecnica de PCR, eletroforese em gel de gradiente desnaturante (DGGE), analise de restricao e sequenciamento direto de DNA, nos analisamos esses genes em 99 tecidos encefalicos de pacientes com DA comprovada histologicamente. Uma amostra de tecido encefalico mostrou uma mutacao no gene PS-1 (His 163 Arg), que mais tarde foi demonstrada ser uma mutacao de celulas germinativas. Nenhuma outra anormalidade de migracao foi demonstrada, em qualquer amostra, nos exons 16 ou 17 do gene APP, nos exons codificadores da PS-1 ou qualquer padrao anormal de digestao pela analise de restricao no gene PS-2...

Frequencia da mutacao delta-mtDNA 4977 e acumulo de placas neuriticas e emaranhados neurofibrilares no giro para-hipocampal humano, no envelhecimento normal e na doenca de Alzheimer / Delta mtDNA 4977 mutational frequence and neuritic plaques and neurofibrillary tangles in human parahippocampal gyrus, in normal aging and Alzheimer's disease

A presenca da mutacao-delecao mtDNA no giro para-hipocampal humano foi investigada em 95 pacientes autopsiados de tres series de origens geograficas distintas, Alemanha, Brasil e Japao, incluindo 70 pacientes sem doencas neuropsiquiatricas e 25 pacientes portadores da doenca de Alzheimer. Somente a serie alema, caracterizada por maiores proporcoes de neuronios medios e grandes, e alta incidencia de placas neuriticas e emaranhados neurofibrilares no giro para-hipocampal, apresentou a delta-mtDNA em niveis detectaveis pela reacao de cadeia da polimerase (PCR). As series brasileira e japonesa, caracterizadas por menores proporcoes de neuronios medios e grandes e baixa incidencia de placas e emaranhados, nao apresentaram niveis detectaveis da alfa-mtDNA. A frequencia f da alfa-mtDNA foi tres vezes menor no grupo de pacientes portadores da doenca de Alzheimer (f=0,12) que no grupo controle (f=0,37) (p=0,03)...