Ancient DNA confirmation of lepromatous leprosy in a skeleton with concurrent osteosarcoma, excavated from the leprosarium of St. Mary Magdalen in Winchester, Hants., UK.

To establish a biological profile and disease aetiologies for one of four burials recovered during a Time Team dig at the St. Mary Magdalen leprosarium, Winchester, UK in AD 2000. Osteological techniques were applied to estimate age at death, biological sex, stature and pathology. Visual assessment of the material was supplemented by radiographic examination. Evidence for leprosy DNA was sought using ancient DNA (aDNA) analysis. The remains are those of a male individual excavated from a west-east aligned grave. The skeleton shows signs of two pathologies. Remodelling of the rhino-maxillary area and degenerative changes to small bones of the feet and reactive bone on the distal lower limbs suggest a multibacillary form of leprosy, whereas the right tibia and fibula show the presence of a primary neoplasm identified as an osteosarcoma. The aDNA study confirmed presence of Mycobacterium leprae in several skeletal elements, and the strain was genotyped to the 3I lineage, one of two main SNP types present in mediaeval Britain and ancestral to extant strains in America. This is a rare documentation of leprosy in association with a primary neoplasm.

Unusual effects of Be doping in the iron-based superconductor FeSe.

Recent superconducting transition temperatures (T c) over 100 K for monolayer FeSe on SrTiO3 have renewed interest in the bulk parent compound. In KCl:AlCl3 flux-transport-grown crystals of FeSe0.94Be0.06, FeSe0.97Be0.03 and, for comparison, FeSe, this work reports doping of FeSe using Be-among the smallest of possible dopants, corresponding to an effective 'chemical pressure'. According to lattice parameter measurements, 6% Be doping shrank the tetragonal FeSe lattice equivalent to a physical pressure of 0.75 GPa. Using this flux-transport method of sample preparation, 6% of Be was the maximum amount of dopant achievable. At this maximal composition of FeSe0.94Be0.06, the lattice unit cell shrinks by 2.4%, T c-measured in the bulk via specific heat-increases by almost 10%, the T c versus pressure behavior shifts its peak [Formula: see text] downwards by ~1 GPa, the high temperature structural transition around T S = 89 K increases by 1.9 K (in contrast to other dopants in FeSe which uniformly depress T S), and the low temperature specific heat γ increases by 10% compared to pure FeSe. Also, upon doping by 6% Be the residual resistivity ratio, ρ(300 K)/ρ(T → 0), increases by almost a factor of four, while ρ(300 K)/ρ([Formula: see text]) increases by 50%.

Mycobacterium tuberculosis ESAT-6 is a leukocidin causing Ca2+ influx, necrosis and neutrophil extracellular trap formation.

Mycobacterium tuberculosis infection generates pulmonary granulomas that consist of a caseous, necrotic core surrounded by an ordered arrangement of macrophages, neutrophils and T cells. This inflammatory pathology is essential for disease transmission and M. tuberculosis has evolved to stimulate inflammatory granuloma development while simultaneously avoiding destruction by the attracted phagocytes. The most abundant phagocyte in active necrotic granulomas is the neutrophil. Here we show that the ESAT-6 protein secreted by the ESX-1 type VII secretion system causes necrosis of the neutrophils. ESAT-6 induced an intracellular Ca(2+) overload followed by necrosis of phosphatidylserine externalised neutrophils. This necrosis was dependent upon the Ca(2+) activated protease calpain, as pharmacologic inhibition prevented this secondary necrosis. We also observed that the ESAT-6 induced increase in intracellular Ca(2+), stimulated the production of neutrophil extracellular traps characterised by extruded DNA and myeloperoxidase. Thus we conclude that ESAT-6 has a leukocidin function, which may facilitate bacterial avoidance of the antimicrobial action of the neutrophil while contributing to the maintenance of inflammation and necrotic pathology necessary for granuloma formation and TB transmission.

Intracerebral transplantation of adult mouse neural progenitor cells into the Niemann-Pick-A mouse leads to a marked decrease in lysosomal storage pathology.

Niemann-Pick disease is caused by a genetic deficiency in acid sphingomyelinase (ASM) leading to the intracellular accumulation of sphingomyelin and cholesterol in lysosomes. In the present study, we evaluated the effects of direct intracerebral transplantation of neural progenitor cells (NPCs) on the brain storage pathology in the ASM knock-out (ASMKO) mouse model of Type A Niemann-Pick disease. NPCs derived from adult mouse brain were genetically modified to express human ASM (hASM) and were transplanted into multiple regions of the ASMKO mouse brain. Transplanted NPCs survived, migrated, and showed region-specific differentiation in the host brain up to 10 weeks after transplantation (the longest time point examined). In vitro, gene-modified NPCs expressed up to 10 times more and released five times more ASM activity into the culture media compared with nontransduced NPCs. In vivo, transplanted cells expressed hASM at levels that were barely detectable by immunostaining but were sufficient for uptake and cross-correction of host cells, leading to reversal of distended lysosomal pathology and regional clearance of sphingomyelin and cholesterol storage. Within the host brain, the area of correction closely overlapped with the distribution of the hASM-modified NPCs. No correction of pathology occurred in brain regions that received transplants of nontransduced NPCs. These results indicate that the presence of transduced NPCs releasing low levels of hASM within the ASMKO mouse brain is necessary and sufficient to reverse lysosomal storage pathology. Potentially, NPCs may serve as a useful gene transfer vehicle for the treatment of CNS pathology in other lysosomal storage diseases and neurodegenerative disorders.

Plutonium-based superconductivity with a transition temperature above 18 K.

Plutonium is a metal of both technological relevance and fundamental scientific interest. Nevertheless, the electronic structure of plutonium, which directly influences its metallurgical properties, is poorly understood. For example, plutonium's 5f electrons are poised on the border between localized and itinerant, and their theoretical treatment pushes the limits of current electronic structure calculations. Here we extend the range of complexity exhibited by plutonium with the discovery of superconductivity in PuCoGa5. We argue that the observed superconductivity results directly from plutonium's anomalous electronic properties and as such serves as a bridge between two classes of spin-fluctuation-mediated superconductors: the known heavy-fermion superconductors and the high-T(c) copper oxides. We suggest that the mechanism of superconductivity is unconventional; seen in that context, the fact that the transition temperature, T(c) approximately 18.5 K, is an order of magnitude greater than the maximum seen in the U- and Ce-based heavy-fermion systems may be natural. The large critical current displayed by PuCoGa5, which comes from radiation-induced self damage that creates pinning centres, would be of technological importance for applied superconductivity if the hazardous material plutonium were not a constituent.

Overexpression of heat-shock proteins reduces survival of Mycobacterium tuberculosis in the chronic phase of infection.

Elevated expression of heat-shock proteins (HSPs) can benefit a microbial pathogen struggling to penetrate host defenses during infection, but at the same time might provide a crucial signal alerting the host immune system to its presence. To determine which of these effects predominate, we constructed a mutant strain of Mycobacterium tuberculosis that constitutively overexpresses Hsp70 proteins. Although the mutant was fully virulent in the initial stage of infection, it was significantly impaired in its ability to persist during the subsequent chronic phase. Induction of microbial genes encoding HSPs might provide a novel strategy to boost the immune response of individuals with latent tuberculosis infection.

Hyperplastic changes within the leptomeninges of the rat and monkey in response to chronic intracerebroventricular infusion of nerve growth factor.

Recombinant human nerve growth factor (rhNGF) was delivered for up to 6 months by continuous intracerebroventricular (i.c.v.) infusion to CD (Sprague-Dawley derived) rats and cynomolgus monkeys. Rats (n = 15/sex/group) received doses of 0 (vehicle), 6, 60, or 300 ng/day; monkeys (n = 5/sex/group) received 0, 0.6, 6, or 60 microg/day of rhNGF. Animals tolerated i.c.v. infusion with no behavioral signs attributable to rhNGF. Body weight was transiently decreased in female rats at the highest dose. At the completion of dosing, histological examination in both species revealed an increase in the thickness of the leptomeninges along the ventral and lateral surfaces of the hindbrain and extending over the dorsal aspect of the spinal cord. The change was present to varying degrees at all doses of rhNGF and tended to be more severe at higher doses. At the light microscopic level, the leptomeninges contained layers of well-differentiated, spindle-shaped cells and a plexus of axonal fibers. Cells were immunoreactive for S-100 protein and were associated with an accumulation of Type IV collagen, suggesting Schwann cell origin. Electron microscopy revealed numerous fine caliber axons ensheathed by the presumptive Schwann cells, with myelination of individual axonal segments. These findings suggest that chronic i.c.v. delivery of rhNGF has stimulated axonal sprouting and secondary hyperplasia of Schwann or Schwann-like support cells within the pia-arachnoid.

Cloning and characterisation of a glutamate dehydrogenase cDNA from tomato (Lycopersicon esculentum L.).

A full-length cDNA (legdh1) has been cloned encoding glutamate dehydrogenase (GDH) from tomato (Lycopersicon esculentum L.). legdh1 is 1568 bp long and contains an open reading frame encoding a 44.8 kDa polypeptide with a putative mitochondrial-matrix-targeting pre-sequence at its N-terminus. Southern analysis indicates the existence of one copy of legdh1 per haploid genome, and no closely related genes were detected by Southern analysis at low stringency. We hypothesise that in tomato, the two GDH subunits may arise from post-transcriptional modifications of a single gene. Northern analysis reveals high expression of legdh1 in roots, lower levels of expression in stems, flowers and leaves, and no detectable expression in fruits. In general, there was no correlation between steady-state mRNA level and protein activity in the tissues analysed, again suggesting the importance of post-transcriptional events in the regulation of GDH. Comparison of cloned plant GDH proteins reveals a high degree of homology throughout the sequence except for a very specific, highly divergent region.

Mineralization of the globus pallidus following excitotoxic lesions of the basal forebrain.

The excitotoxin N-methyl aspartic acid was injected into the rat nucleus basalis to destroy basal forebrain cholinergic (BFC) neurons. In long-term survival experiments (up to 11 months post-lesion), conspicuous mineralized deposits were found in the globus pallidus and to a lesser extent in the thalamus. Deposits stained in a manner consistent with a composition of calcium and iron. Typically, deposits were absent from the center of the injection site, where BFC cell loss was most severe, but were present within the ventral and lateral globus pallidus where there was substantial sparing of BFC neurons. The similarity of this pathology to basal ganglia calcification and its relationship to Alzheimer's Disease and Down's syndrome is discussed.

Recombinant human ciliary neurotrophic factor stimulates the metabolic activity of SH-SY5Y cells as measured by a cytosensor microphysiometer.

Information on the transmembrane signaling events and subsequent biochemical processes initiated by ciliary neurotrophic factor (CNTF) receptor activation in neurons is lacking. SH-SY5Y cells, a human neuroblastoma cell line expressing CNTF receptors, were used to study metabolic changes associated with functional ligand-receptor interactions. Real-time measurements quantifying the rate of extracellular acidification by SH-SY5Y cells (a measure of metabolic activity) were made using a silicon-based cytosensor. Application of recombinant human CNTF (rhCNTF) to resting SH-SY5Y cells increased their acidification rate in a concentration and time-dependent manner with an apparent EC50 of 60 ng/ml. Pretreatment of cells with phosphatidylinositol-specific phospholipase C (PI-PLC) prevented the CNTF, but not an NGF-stimulated increase in acidification rate. Collectively, these results demonstrate that: (1) SH-SY5Y cells express functional CNTF receptors; and (2) the initial signal transduction mechanism activated by the CNTF receptor in SH-SY5Y cells is distinct from that activated by the NGF receptor; however, both may ultimately stimulate the same downstream biochemical messengers to increase cellular metabolism.

Role of Ethylene in the Germination of the Hemiparasite Striga hermonthica.

Seed germination of the hemiparasitic angiosperm Striga hermonthica (Del.) Benth is elicited by compounds present in the root exudates of the host plant. Although a variety of compounds can substitute for the host-derived signal, the mechanism through which these act is unknown. In the present study, an inhibitor of ethylene biosynthesis, aminoethoxyvinyl glycine, was found to inhibit germination. Addition of an intermediate in ethylene biosynthesis, 1-aminocyclopropane-1-carboxylic acid, was found to override this inhibition and to act as a substitute for the host-derived signal. 2,5-Norbornadiene, an inhibitor of ethylene action, was also found to inhibit germination. Ethylene is rapidly produced by Striga seeds after treatment with host root exudates. These results are consistent with a model for Striga seed germination in which host-derived signals and other compounds act by eliciting the synthesis of ethylene and in which ethylene itself initiates the biochemical changes leading to germination.

Prophylactic cranial irradiation for limited non-small cell lung cancer.

Seventy-three patients with biopsy-proven limited non-small cell lung cancer (NSCLC) were entered on a combined modality study at the University of Washington. Seventy-five percent (55 of 73) of the patients had a histologic diagnosis of adenocarcinoma or large cell carcinoma, whereas 25% (18 of 73) had squamous cell carcinoma. After two cycles of chemotherapy, patients without evidence of progressive disease received prophylactic cranial irradiation (PCI) and chest radiotherapy, followed by two additional cycles of chemotherapy. Brain computed tomography (CT) scans were performed at 3-month intervals after completion of therapy in all patients, and were additionally performed whenever signs or symptoms developed suggesting neurologic dysfunction or recurrent brain disease. Sixty-five patients were treated with PCI. No clinical or CT evidence of recurrence in the brain has developed in patients who completed PCI. PCI appears to be effective in greatly reducing the incidence of brain relapse in patients with limited NSCLC.

Fast neutron radiotherapy--an overview.

Fast neutron radiotherapy has been employed worldwide in the last two decades in the treatment of a variety of advanced malignancies. The basic principles behind neutron treatment will be summarized in this article, along with results to date of a number of clinical trials evaluating the effectiveness and toxicity of neutron radiotherapy.

Choroid plexus carcinoma of the fourth ventricle. Report of a case in an infant.

An 8-month-old female infant with choroid plexus carcinoma presenting in the fourth ventricle is described. The patient was initially treated with combination chemotherapy due to her young age, but developed local tumor recurrence 13 months after beginning treatment. Staging studies to detect tumor seeding of the cerebrospinal fluid were negative, and the patient received posterior fossa radiotherapy. Although the primary tumor regressed with radiation treatment, widespread disease throughout the spinal subarachnoid space subsequently developed. Choroid plexus carcinoma has a significant tendency to metastasize through the cerebrospinal fluid, and treatment of the entire neuraxis should be strongly considered for patients with this disease.

CT manifestation in the pararectal area following fast neutron radiotherapy.

Sixty-four pelvic CT studies of 38 patients with pelvic malignancies were obtained at various times (21 to 678 days) following fast neutron radiotherapy. Multiple scans (2 to 3) were available in 19 cases. Fractionated neutron radiotherapy was delivered over four to seven weeks to a total dose of 54 Gy to 60 Gy photon equivalent. Radiation-induced changes on CT were symmetrical thickening of the perirectal fascia (87%), widening of the presacral space (82%), and thickening of the walls of the bladder and rectum (21% and 11%, respectively). Serial CT studies showed that these changes became stable after 12 weeks from radiotherapy in 13 of 14 patients (93%). CT manifestations of radiation-induced changes in the pararectal area were rather regular and would be easy to differentiate from local recurrence or secondary disease.

Gas exchange characteristics of the sorghum-striga host-parasite association.

Gas exchange characteristics are reported for both members of the sorghum-Striga host-parasite association. Both Striga hermonthica (Del.) Benth and Striga asiatica (L.) Kuntze had transpiration rates considerably in excess of those of sorghum (Sorghum bicolor (L.) Moench, cv CSH1). Stomatal conductance in both Striga spp. showed little response to periods of darkness and moderate water stress. Low rates of net CO(2) fixation and high rates of dark respiration led to no net daily (24 hours) C gain, and Striga would appear to be reliant on its host for photosynthate. Infection of sorghum plants with either S. hermonthica or S. asiatica reduced host photosynthetic capacity. Infected sorghum plants were also more prone to water stress, but reduced rates of CO(2) fixation could not be accounted for in terms of lower stomatal conductance. Lower stomatal conductances were associated with an increase in water use efficiency (WUE) in uninfected sorghum; however, Striga-infected sorghum plants had lower WUE than those of uninfected plants. We suggest that Striga exerts a specific effect on processes affecting C acquisition in sorghum leaves. The water relations of S. hermonthica and S. asiatica are not characteristic of plants growing in semiarid environments and are more likely to reflect the nature of the parasitic life-style. Despite transfer of water and solutes from host to parasite, the reduction in C fixation observed in infected sorghum plants appears to be the major determinant of growth reductions observed in sorghum supporting Striga.

Transthoracic right atrial cannulation for total parenteral nutrition- case report.

A man with severe peripheral vascular disease and requiring total parenteral nutrition because of short bowel syndrome was referred because a central venous catheter could not be inserted by conventional techniques. A right thoracotomy was performed and a Hickman catheter inserted via the right atrial appendage into the right atrium. This catheter was used for a total of seven months for total parenteral nutrition. For the last two months of this time, the patient was maintained at home on a Home Parenteral Nutrition Programme. After four months of total parenteral nutrition the patient developed recurrent fevers and the catheter was found to have migrated from the right atrium into the pulmonary artery. The catheter was resited under X-ray control and used for a further three months until the recurrence of fever and dyspnoea heralded the onset of septic pulmonary emboli resulting in his death.

Rubber sump drainage of enterocutaneous fistulae.

Effective sump drainage of high enterocutaneous fistulae, together with alimentary rest and total parenteral nutrition, is now an integral part of the modern management of patients with this condition. The low tissue reactivity of the plastic and polymer materials currently used in most drainage tubes appears however, to be counterproductive to the establishment of a discrete fistula track and control of the fistula. A case is made for the use of red rubber sump drains for enterocutaneous fistulae. The greater tissue reactivity of rubber is reviewed and confirmed by animal experimentation. A method of rubber sump drainage of enterocutaneous fistulae developed during the management of 83 such fistulae is described.

Early detection of breast cancer.

One thousand referred patients with breast symptoms, a history of breast disease, or a family history of breast carcinoma have been examined at a suburban breast clinic. This is a preliminary survey of the data and results so far obtained from the initial examination of these patients. The examination consisted of history-taking by means of a questionnaire, physical examination of the breasts, thermography and xeromammography. A report correlating the findings was forwarded to the referring doctor in each case and any necessary action was indicated in the report. Seventy-eight patients were thought to require biopsy of a suspicious lump or mammographic lesion (7.8% of the total number); 76 patients actually had biopsies, and 18 patients were found to have carcinoma of the breast (23.8% of the total number of patients who had biopsies). The overall incidence of carcinoma in the 1000 women attending the clinic was 1.8% at the initial visit. While general population screening for breast carcinoma may be hard to justify, from both the detection rate and an economic point of view, it would appear from our experience to date that screening of selected high-risk referred patients is justified and, in fact, desirable.