Endoneurial pathology of the needlestick-nerve-injury model of Complex Regional Pain Syndrome, including rats with and without pain behaviors.

Current rodent models of neuropathic pain produce pain hypersensitivity in almost all lesioned animals and not all identified experimental effects are pain specific. 18G needlestick-nerve-injury (NNI) to one tibial nerve of outbred Sprague-Dawley rats models the phenotype of Complex Regional Pain Syndrome (CRPS), a post-traumatic neuropathic pain syndrome, leaving roughly half of NNI rats with hyperalgesia. We compared endoneurial data from these divergent endophenotypes searching for pathological changes specifically associated with pain-behaviors. Tibial, sural, and common sciatic nerves from 12 NNI rats plus 10 nerves from sham-operated controls were removed 14 days post-surgery for morphometric analysis. PGP9.5(+) unmyelinated-fibers were quantitated in plantar hindpaw skin. Distal tibial nerves of NNI rats had endoneurial edema, 30% fewer axons, twice as many mast cells, and thicker blood-vessel walls than uninjured tibial nerves. However the only significant difference between nerves from hyperalgesic versus non-hyperalgesic NNI rats was greater endoneurial edema in hyperalgesic rats (p < 0.01). We also discovered significant axonal losses in uninjured ipsilateral sural nerves of NNI rats, demonstrating spread of neuropathy to nearby nerves formerly thought spared. Tibial and sural nerves contralateral to NNI had significant changes in endoneurial blood-vessels. Similar pathological changes have been identified in CRPS-I patients. The current findings suggest that severity of endoneurial vasculopathy and inflammation may correlate better with neuropathic pain behaviors than degree of axonal loss. Spread of pathological changes to nearby ipsilateral and contralateral nerves might potentially contribute to extraterritorial pain in CRPS.

Effects of distal nerve injuries on dorsal-horn neurons and glia: relationships between lesion size and mechanical hyperalgesia.

Penetrating limb injuries are common and usually heal without long-lasting effects, even when nerves are cut. However, rare nerve-injury patients develop prolonged and disabling chronic pain (neuralgia). When pain severity is disproportionate to severity of the inciting injury, physicians and insurers may suspect exaggeration and limit care or benefits, although the nature of the relationship between lesion-size and the development and persistence of neuralgia remains largely unknown. We compared cellular changes in the spinal dorsal-horn (the initial CNS pain-processing area) after partial or total tibial-nerve axotomies in male Sprague-Dawley rats to determine if these changes are proportional to the numbers of peripheral axons cut. Unoperated rats provided controls. Plantar hind-paw responses to touch, pin, and cold were quantitated bilaterally to identify hyperalgesic rats. We also compared data from nerve-injured rats with or without hyperalgesic responses to mechanical hind-paw stimulation to evaluate concordance between pain behaviors and dorsal-horn cellular changes. Hyperalgesia was no less prevalent or severe after partial than after total axotomy. L(5) spinal-cord sections from rats killed 7 days postoperatively were labeled for markers of primary afferents (substance P calcitonin gene-related peptide isolectin B4, gamma aminobutyric acid, and glial fibrillary acidic protein), then labeled cells were stereologically quantitated in somatotopically defined dorsal-horn regions. Total axotomy reduced markers of primary afferents more than partial axotomy. In contrast, GABA-immunoreactive profiles were similarly reduced after both lesions, and in rats with sensory loss versus hyperalgesia. Numbers of GFAP-immunoreactive astrocytes increased independently of lesion size and pain status. Small nerve injuries can thus have magnified and disproportionate effects on dorsal-horn neurons and glia, perhaps providing a biological correlate for the disproportionate pain of post-traumatic neuralgias (including complex regional pain syndrome-I) that follow seemingly minor nerve injuries. However, the presence of similar dorsal-horn changes in rats without pain behaviors suggests that not all transcellular responses to axotomy are pain-specific.

Treatment of patients with asymmetries using asymmetric mechanics.

The orthodontic correction of dental asymmetries is often considered a difficult and challenging process, primarily because of misdiagnosis and poorly planned treatment mechanics. A careful differential diagnosis together with a thorough treatment plan can ensure successful treatment outcomes in the management of these malocclusions. In this article, key elements of differential diagnosis and treatment planning are reviewed. Different treatment strategies, including an analysis of the side effects associated with the use of these appliance systems, are described. This approach will help the clinician to best adapt treatment mechanics to a given malocclusion.

Combined orthodontic and prosthetic therapy. Special considerations.

The adult patient may present with a complex medical and dental history. Comprehensive treatment planning with appropriate consultation among the restorative dentist and other dental specialists is a prerequisite for a successful treatment outcome. This article reviewed the role of orthodontic treatment in the restoration of the partially edentulous patient. The case illustrations presented show the value of collaborative efforts in a multidisciplinary team approach. Although there may be initial reluctance on the part of some patients to seek multiple specialty consultations, the rewards are great for the patient and the dentists who subscribe to this treatment philosophy.

Stroke as a late sequela of cranial irradiation for childhood brain tumors.

Cerebrovascular disease involving large and medium-size vessels is thought to be an uncommon sequela of treatment of childhood brain tumors. We reviewed 11 children who developed cerebrovascular disease manifested by strokes or transient ischemic attacks 6 months to 4 years after treatment of brain tumors, while their tumors were in remission. All had received radiation therapy, and seven had received chemotherapy. One child died of acute bilateral cerebral infarctions due to carotid occlusion on one side and marked stenosis on the other 2 years after receiving radiation therapy for an incompletely resected craniopharyngioma. Pathologically, there was marked subendothelial fibrosis of the vessels of the circle of Willis, with inflammatory changes surrounding some of the vessels. In addition to the widely recognized small-vessel damage caused by radiation and chemotherapy in children (mineralizing microangiopathy), damage to medium and large intracranial vessels may result in late sequelae, manifested by stroke or transient ischemic attacks.

The protection of invertebrates, fish, and vascular plants against inorganic mercury poisoning by sulfur and selenium derivatives.

Protection of organisms against mercury (Hg) poisoning is most commonly associated with the antagonistic effects of selenium (Se)-compounds against mercury alkyls in higher animals. This study shows that there is no consistent difference among Periodic Group VIA derivatives including S(IV), S(II) organic, Se (IV), Se (II) organic, possibly Se VI, as well and Te (IV) in their ability to protect against mercury poisoning. The organisms used in assays were: Coleus explants (leaf abscission); turnip (germination); pea (growth inhibition and Hg uptake); a planarian (regeneration); the brineshrimp (excystment, phototaxy); the mealworm larva Tenebrio (metamorphosis) and the fish "tilapia" (survival, Hg uptake). Thiamine was the most effective of the Group VIA derivatives against the widest spectrum of organisms and test systems. In planarian regeneration, it was active where S and Se compounds failed. The most unexpected observation was the hastening of insect metamorphosis by HgCl2 and the enhancement of that effect by thiamine.

Ultrastructural localization of uranium biosorption in Penicillium digitatum by stem X-ray microanalysis.

When Penicillium digitatum Saccardo cultures are exposed to aqueous solutions containing soluble uranium salts, considerable amounts of this element are accumulated in the fungal mycelium. The accumulated uranium is retained after thorough rinsing with distilled water but is removed by alkali carbonate solutions. Analysis of thick sections (0.5 microm) of the fungal hyphae with TEM, after incubation in UO(2)Cl(2) solutions of varying concentrations under both light and dark conditions, revealed conspicuous crystal-like deposits in UO(2)Cl(2)-exposed hyphae, but none in the control hyphae. Thick sections were necessary for crystal visualization. Using energy-dispersive X-ray analysis, uranium was detected as the only heavy element in these crystals. Uranium crystal biosorption was localized on the outside surface of the hyphal cell wall (following short exposures to relatively low uranium concentrations) or inside the cell wall (following long exposure to relatively high uranium concentrations). In some cases, crystal-like deposits of uranium salts were located on the outside surface as well as inside the cell.

The prognostic implication of hypercholesterolemia in infants and children with hepatoblastoma.

This paper reports the relationship between serum cholesterol level and hepatoblastoma in nine patients. Four of the nine patients had a high (417-544 mg%) serum cholesterol. All of these patients were less than one year of age and had tumors of the epithelial type. Three of the infants died soon after being diagnosed. Three patients had a moderately elevated serum cholesterol (206-249mg%). One underwent primary hepatic resection. Two had nonresectable tumors that became resectable after chemotherapy. One died secondary to pulmonary metastases. The remaining two patients had a normal serum cholesterol level prior to treatment. Both patients had hepatic resection after chemotherapy and had no evidence of increased cholesterol postoperatively and are alive six years after diagnosis. From these data, it is suggested that the pretreatment level of serum cholesterol may be of prognostic significance in infants and children with hepatoblastoma.

Short-term and long-term clinostat and vibration-induced biochemical changes in dwarf marigold stems.

Stems of 21-day dwarf Marigold plants cultivated on the clinostat were compared with plants cultivated on vertical axis rotators ("vibrational controls") and stationary controls for long-term changes in cell wall composition. Stems of 21-day plants grown under stationary conditions and subsequently exposed to the clinostat for 24 hours were also analyzed. Among the long-tern markers, calciun, lignin, and protein-bound hemicellulose (possibly cell wall glycoprotein) clearly differentiated the effects of vibration from those of the clinostat. Short-term differential responses included rate of ethylene production, nastic movement and peroxidase activity of the cell wall, but not of the protoplast.

An extraterrestrial habitat on Earth: the algal mat of Don Juan [correction of Jaun] Pond.

On the edge of Don Juan Pond in the Wright Valley of Antarctica lies a mat of mineral and detritus cemented by organic matter. In spite of a CaCl2 concentration of about 33% (w/v), the mat contains Oscillatoria and other cyanobacteria, unicellular forms, colonial forms rich in carotenoids, and diatoms. Bacteria are rare; fungal filaments are not. Oscillatoria showed motility but only at temperatures <10 degrees C. Acetone extracts of the mat and nearby muds yielded visible spectra similar to those of laboratory grown O. sancta, with 50- to 70-fold molar ratio of chlorophyll a to b. Although rare, tardigrades were also found. The algal mat had enzymatic activities characteristic of peroxidase, catalase, dehydrogenase, and amylase. Cellulose, chitin, protein, lipid and ATP were present. Previously, algae in the Wright Valley have been described in melt water, not in the brine itself. Wright Valley has been used as a near sterile Martian model. It obviously contains an array of hardy terrestrial organisms.

Removal of Uranium(VI) from Solution by Fungal Biomass and Fungal Wall-Related Biopolymers.

Penicillium digitatum mycelium can accumulate uranium from aqueous solutions of uranyl chloride. Azide present during the uptake tests does not inhibit the process. Killing the fungal biomass in boiling water or by treatment with alcohols, dimethyl sulfoxide, or potassium hydroxide increases the uptake capability to about 10,000 parts per million (dry weight). Formaldehyde killing does not enhance the uranium uptake. The inference that wall-binding sites were involved led to the testing of uranium uptake by chitin, cellulose, and cellulose derivatives in microcolumns. All were active, especially chitin.

Mercury content of equisetum plants around mount st. Helens one year after the major eruption.

The mercury content of young Equisetum plants collected around Mount St. Helens was higher in the direction of Yakima and Toppenish, Washington (northeast to east-northeast), than at any other compass heading and was about 20 times that measured around Portland, Oregon. The increase in substratum mercury was not as pronounced as that in plants but was also higher toward the northeast, the direction taken by the May 1980 volcanic plume.

Enzyme-mimicking properties of silicates and other minerals.

The adsorptive and/or catalytic properties of clays, silicates in general, and other minerals are well known. More recently, their probable role in prebiotic syntheses of bio-organic compounds has become a matter of record. We demonstrate that, in addition to their role in de novo formation of important biomolecules, clays, micas, fibrous silicates and other minerals mimic the activities of contemporary enzymes including oxidases, esterases, phosphatases and glucosidases. The existence of such capabilities in substances likely to be represented on the surfaces of Earth-like planets may offer a challenge to the technology and design of remote life detection systems which must then distinguish between bona fide biological chemistry and mineral-base pseudometabolism. It also raises questions about the importance of mineral surfaces in post-mortem transformations of organic metabolites in our own biosphere.

Effects of salts and temperatures on post-irradiation growth of Penicillium exposed to ultraviolet.

The growth of Penicillium notatum colonies after UV irradiation of dried mycelium or spores was studied in relation to post-irradiation temperature and salt environment. Dried mycelium and spores behaved differently with respect to sensitivity to temperature, salts and UV, especially the latter. Threshold inhibitory doses for spores were modified markedly either at 4 degrees C or in magnesium and calcium chlorides. It is suggested that these temperature and salt effects are related to prevention of photochemical membrane damage.

Further studies on the environmental capabilities of fungi: interactions of salinity, ultraviolet irradiation, and temperature in Penicillium.

A strain of Penicillium currently under study in this laboratory has revealed some unexpected features in response to salts and UV-radiation. These responses include: (a) stimulation of growth by one molar NaCl and KCl at 23 degrees but not at 5 degrees C, (b) total inhibition by LiCl and RbCl at 5 degrees but not at 23 degrees C, (c) stimulation by 254 nm radiation (2 x 10(6) erg cm-2 min-1, 30-90 minutes) at 23 degrees and 5 degrees C, and (d) very elevated growth rates at 23 degrees after incubation at 5 degrees C.

Gravity and land plant evolution: experimental induction of lignification by simulated hypergravity and water stress.

The ability to lignify cell walls has long been associated with the upright condition and sometimes massive body of the land plant, and gravitational regulation of lignification has been demonstrated by clinostat/centrifuge techniques. Previous centrifuge experiments were conducted using plants in the early stages of growth, hence already programmed for immediate lignification. Elodea, an aquatic Angiosperm long believed to be incapable of lignification synthesizes lignin or lignin-like phenolics during 6 days incubation at 80 g . Yields in some cases exceeded stationary controls by as much as 300%. Supporting data were obtained using bean cotyledons. Exposure to 80 g induced lignification in extra-vascular tissue along with vascular lignin deposits. The loss of a buoyant medium also deprived early land plants of unlimited water: the lignin content in cucumber seedlings grown in water is reduced 50% or more relative to pot grown plants but water stress elevates lignin content as much as 100%. This suggests that the water status of the land environment may have provided a "backup" or "reinforcement" signal for an adaptive axial support system.

Growth of fungi in NaCl-MgSO4 brines.

Long-term studies have shown that common fungi of the Penicillium-Aspergillus group can be grown in a variety of brines or on moist salt crystals, simulating a range of natural terrestrial habitats such as salt flats, or special water-bodies such as the Dead Sea. In general, salt media rich in KCl are favored over other alkali halides; the media become more selective as the salt concentration rises and nutrient requirements become more complex. We here demonstrate that media which resemble the Dead Sea salt mix can support the growth of selected fungal strains, even in the absence of reduced organic nutrients other than glucose. Such media may serve as models for localized microhabitats on Mars.

Gravity as a biochemical determinant.

The existence of obvious morphological and physiological changes in living systems exposed to altered gravity immediately informs us that prior changes have taken place in the chemistry of exposed cells, tissues and organs. These changes include transients that return more or less promptly to the norm when the system is restored to the terrestrial g-field. For example, altered serum hormone and electrolyte levels in man, which appear to reflect successful adaptation to the conditions of orbital weightlessness, disappear shortly after return to Earth. Other changes--in mineral and protein constituents of the skeletal system in man, and cell wall composition in plants--are more persistent or even permanent. Hypogravitational departures from the norm include not only "weightlessness" as achieved in orbit, but also experimental modes of compensation, on the clinostat or by flotation. These techniques are useful in the study of hypogravity but cannot replace fully the weightless environment. Plant ethylene and peroxidase both increase under orbital, clinostat and/or flotation conditions whereas 3-phosphoglyceraldehyde-dehydrogenase increases under orbital but not clinostat conditions; cytochrome reductase and malic dehydrogenase levels are affected by the clinostat, but not by actual weightless conditions. How do the altered organismal biochemistries induced by the centrifuge and the clinostat relate to one another? Does gravity operate on living systems as a continuous variable from 0 to superterrestrial values, or do deviations from g(earth) generate non-uniform, discontinuous stress responses, irrespective of sign? In plants, measurements of wall lignin content and peroxidase activity yield opposite answers. Given the limited data so far available we will consider the meaning of these contradictions.