Sorption/Desorption of lincomycin from three arid-region soils.

The antibiotic lincomycin is commonly found in treated municipal waste water and in waste from swine and poultry production. Environmental disposal of these wastes has the potential to introduce a significant mass of lincomycin into the ecosystem. In the present study, a series of sorption and desorption experiments were conducted to determine the potential mobility of lincomycin in soils from arid environments. Sorption and desorption isotherms were obtained for lincomycin using three different soils. Isotherms were fit to the Freundlich equation. Adsorption of lincomycin was found to have a of 11.98 for a biosolid-treated soil (1.58% OC) and a of 210.15 for a similar unamended soil (1.42% OC). It was also found that for a low-organic-content soil the was 5.09. The differences in adsorption can be related to the soil pH and the pKa of lincomycin (7.5-7.8). When the soil solution pH is below the pKa, the cationic species of lincomycin dominates, resulting in increased water solubility. Interaction with the cation exchange complex is minimal due to a high solution cation concentration (Ca and Na). Desorption isotherms also indicate that when the solution pH is lower than the pKa, retention of lincomycin is reduced. Our results indicate that the mobility of lincomycin in these arid region soils is dependent on soil pH.

Duration of preclinical rheumatoid arthritis-related autoantibody positivity increases in subjects with older age at time of disease diagnosis.

OBJECTIVES: To investigate factors that may influence the prevalence and timing of appearance of rheumatoid factor (RF) and anti-cyclic citrullinated peptide (anti-CCP) antibodies during the preclinical phase of rheumatoid arthritis (RA) development. METHODS: 243 serial prediagnosis serum samples from 83 subjects with RA were examined for the presence of RF and anti-CCP antibodies. RESULTS: Of the 83 cases, 47 (57%) and 51 (61%) subjects had at least one prediagnosis sample positive for RF or anti-CCP, respectively. Gender and race were not significantly associated with the prevalence or timing of preclinical antibody appearance. Preclinical anti-CCP positivity was strongly associated with the development of erosive RA (odds ratio = 4.64; 95% confidence interval 1.71 to 12.63; p<0.01), but RF was not (p = 0.60). Additionally, as age at the time of diagnosis of RA increased the duration of prediagnosis antibody positivity for RF and anti-CCP increased, with the longest duration of preclinical antibody positivity seen in patients diagnosed with RA over the age of 40. In no subjects did symptom onset precede the appearance of RF or anti-CCP antibodies. CONCLUSIONS: The period of time that RF and anti-CCP are present before diagnosis lengthens as the age at the time of diagnosis of RA increases. This finding suggests that factors such as genetic risk or environmental exposure influencing the temporal relationship between the development of RA-related autoantibodies and clinically apparent disease onset may differ with age.

Two centuries of the Scandinavian wolf population: patterns of genetic variability and migration during an era of dramatic decline.

The grey wolf (Canis lupus) was numerous on the Scandinavian peninsula in the early 19th century. However, as a result of intense persecution, the population declined dramatically and was virtually extinct from the peninsula by the 1960s. We examined historical patterns of genetic variability throughout the period of decline, from 1829 to 1979. Contemporary Finnish wolves, considered to be representative of a large eastern wolf population, were used for comparison. Mitochondrial DNA (mtDNA) variability among historical Scandinavian wolves was significantly lower than in Finland while Y chromosome variability was comparable between the two populations. This may suggest that long-distance migration from the east has been male-biased. Importantly though, as the historical population was significantly differentiated from contemporary Finnish wolves, the overall immigration rate to the Scandinavian peninsula appears to have been low. Levels of variability at autosomal microsatellite loci were high by the early 1800s but declined considerably towards the mid-20th century. At this time, approximately 40% of the allelic diversity and 30% of the heterozygosity had been lost. After 1940, however, there is evidence of several immigration events, coinciding with episodes of marked population increase in Russian Karelia and subsequent westwards migration.

Multifocal, metachronous, giant cell tumor of the lower limb.

A case of multifocal giant cell tumor in a skeletally immature male with documented metachronous disease of the lower limb is described followed by a review of the literature including treatment options and their outcomes.

Multiple protein differences distinguish clam leukemia cells from normal hemocytes: evidence for the involvement of p53 homologues.

In coastal locations, marine invertebrates, primarily molluscs, develop fatal leukemias in their blood or hemolymph. In the clam Mya arenaria, non-adhesive, mitotic, spherical leukemia cells replace adhesive, motile, normal hemocytes as leukemia progresses. End-stage leukemia cells express a unique antigen, IE10, while normal cells express the 2A4 marker. The goals of this work were to further differentiate the normal and leukemia specific antigens relative to protein structure, determine if other protein distinctions exist, and examine p53 gene family expression in both cell types. Recognized by the monoclonal antibody 2A4, normal cells express a 185-kDa glycoprotein that may have multiple forms. Detected by the monoclonal antibody 1E10, leukemic cells express a very hydrophobic 252-kDa glycoprotein that is likely to be a transmembrane protein with spectrin/dystrophin-like characteristics. After normalization to the major cytoskeletal protein actin, sodium dodecyl sulfate-polyacrylamide gel electrophoresis reveals major distinguishing protein and glycoprotein differences between the two cell types. Most obvious is the near-absence of tubulin in the non-mitotic normal hemocytes. We have also characterized the expression of p53 gene family members in normal and end-stage leukemia cells, finding shifts in expression of the p53 gene homologues p73 and p97 coincident with leukemia-specific protein synthesis.

Exposure to ultraviolet radiation (290-400 nm) causes oxidative stress, DNA damage, and expression of p53/p73 in laboratory experiments on embryos of the spotted salamander, Ambystoma maculatum.

Developing embryos of the spotted salamander, Ambystoma maculatum, exposed to ultraviolet radiation (UVR; 290-400 nm) in the laboratory show a significant sensitivity to UVB (290-320 nm) radiation. Embryos in laboratory experiments exhibited significant DNA damage during exposures to UVR despite a significant increase in the production of the protective pigment melanin in response to UVR exposure. DNA damage occurs as a result of both the direct effects of exposure to UVR, and the indirect effects are mediated by the production of reduced oxygen intermediates. The production of reactive oxygen species initiates the expression of p53/p73 that leads to either DNA repair or apoptosis. When similar experiments are conducted on salamander embryos exposed to solar UVR in vernal pools, the embryos show significantly less sensitivity and higher survivorship. The differences between laboratory and field experiments are a result of the attenuation of UVR caused by the accumulation of dissolved organic carbon within the pools of these wooded areas. These findings suggest that northeastern populations of spotted salamanders are sensitive to UVR but are not significantly affected by present-day irradiances of UVR in the field. These results do suggest that continued decreases in stratospheric ozone over temperate latitudes have the potential to affect spotted salamanders in their natural habitats.

Cutaneous botryomycosis of the cervicofacial region.

BACKGROUND: Botryomycosis is a rare, chronic, bacterial infection of insidious onset involving the integument or viscera that often mimics actinomycosis or a deep fungal infection. The pathogenesis is thought to be a symbiotic relationship between the host and the infecting organism. METHODS: Case report of a patient with a chronic infection involving the cervicofacial region diagnosed as cutaneous botryomycosis arising from a chronic osteomyelitis of the mandible. The diagnosis was based on the chronicity of the infection along with the identification of botryomycotic (bacteria-containing) granules on histopathologic examination. Special stains excluded fungi and mycobacterium. Cultures identified the offending bacteria, and antibiotic therapy was initiated on the basis of the sensitivities, resulting in resolution of this chronic infectious process. A review of the English language literature revealed that this is the first case of cutaneous botryomycosis arising from a chronic osteomyelitis of the mandible. RESULTS: Medical therapy proved curative at 14 months follow-up. Surgery was performed for diagnostic purposes only. CONCLUSIONS: Botryomycosis is exceedingly rare in the head and neck, and consideration of this entity in the differential diagnosis is critical to the diagnosis. The mainstay of therapy is medical with surgery reserved for biopsy and/or excision of persistent disease. Published 2001 John Wiley & Sons, Inc.

Expression of homologues for p53 and p73 in the softshell clam (Mya arenaria), a naturally-occurring model for human cancer.

Homologues for human p53 (Hsp53) and p73 (Hsp73) genes were cloned and expression patterns for their corresponding proteins analysed in tissues from normal and leukemic softshell clams (Mya arenaria). These are the first structural and functional data for p53 and p73 cDNAs and gene products in a naturally occurring, non-mammalian disease model. Core sequence of the predicted clam p53 (Map53) and p73 (Map73) proteins is virtually identical and includes the following highly conserved regions: the transcriptional activation domain (TAD), MDM2 binding site, ATM phosphorylation site, proline rich domain, DNA binding domains (DBDs) II-V, nuclear import and export signals and the tetramerization domain. The core sequence is a structural mosaic of the corresponding human proteins, with the TAD and DBDs resembling Hsp53 and Hsp73, respectively. This suggests that Map53 and Map73 proteins may function similarly to human proteins. Clam proteins have either a short (Map53) or long (Map73) C-terminal extension. These features suggest that Map53 and Map73 may be alternate splice variants of a p63/p73-like ancestral gene. Map73 is significantly upregulated in hemocytes and adductor muscle from leukemic clams. In leukemic hemocytes, both proteins are absent from the nucleus and sequestered in the cytoplasm. This observation suggests that a non-mutational p53/p73-dependent mechanism may be involved in the clam disease. Further studies of these gene products in clams may reveal p53/p73-related molecular mechanisms that are held in common with Burkitt's lymphoma or other human cancers.

Genetic variation and population structure in Scandinavian wolverine (Gulo gulo) populations.

Wolverine (Gulo gulo) numbers in Scandinavia were significantly reduced during the early part of the century as a result of predator removal programmes and hunting. Protective legislation in both Sweden and Norway in the 1960s and 1970s has now resulted in increased wolverine densities in Scandinavia. We report here the development of 15 polymorphic microsatellite markers in wolverine and their use to examine the population sub-structure and genetic variability in free-ranging Scandinavian wolverine populations as well as in a sample of individuals collected before 1970. Significant subdivision between extant populations was discovered, in particular for the small and isolated population of southern Norway, which represents a recent recolonization. Overall genetic variability was found to be lower than previously reported for other mustelids, with only two to five alleles per locus and observed heterozygosities (H(O)) ranging from 0.269 to 0.376 across the examined populations, being lowest in southern Norway. Analysis of the mitochondrial DNA control region revealed no variation throughout the surveyed populations. As the historical sample did not show higher levels of genetic variability, our results are consistent with a reduction in the genetic variation in Scandinavian wolverines that pre-dates the demographic bottleneck observed during the last century. The observed subdivision between populations calls for management caution when issuing harvest quotas, especially for the geographically isolated south Norwegian population.

Oxidative stress, DNA damage and p53 expression in the larvae of atlantic cod (Gadus morhua) exposed to ultraviolet (290-400 nm) radiation.

Decreases in stratospheric ozone levels from anthropogenic inputs of chlorinated fluorocarbons have resulted in an increased amount of harmful ultraviolet-B (UVB, 290-320 nm) radiation reaching the sea surface in temperate latitudes (30-50 degrees N). In the Gulf of Maine, present-day irradiances of ultraviolet-A (UVA, 320-400 nm) radiation can penetrate to depths of 23 m and UVB radiation can penetrate to depths of 7-12 m, where the rapidly developing embryos and larvae of the Atlantic cod (Gadus morhua) are known to occur. Laboratory exposures of embryos and larvae of Atlantic cod to ultraviolet radiation (UVR) equivalent to a depth of approximately 10 m in the Gulf of Maine resulted in significant mortality of developing embryos and a decrease in standard length at hatching for yolk-sac larvae. Larvae at the end of the experimental period also had lower concentrations of UVR-absorbing compounds and exhibited significantly greater damage to their DNA, measured as cyclobutane pyrimidine dimer formation, after exposure to UVB radiation. Larvae exposed to UVB radiation also exhibited significantly higher activities and protein concentrations of the antioxidant enzyme superoxide dismutase and significantly higher concentrations of the transcriptional activator p53. p53 is expressed in response to DNA damage and can result in cellular growth arrest in the G1- to S-phase of the cell cycle or to programmed cell death (apoptosis). Cellular death caused by apoptosis is the most likely cause of mortality in embryos and larvae in these laboratory experiments, while the smaller size at hatching in those larvae that survived is caused by permanent cellular growth arrest in response to DNA damage. In addition, the sub-lethal energetic costs of repairing DNA damage or responding to oxidative stress may also contribute to poor individual performance in surviving larvae that could also lead to increases in mortality. The irradiances of UVB radiation that elicit these responses in cod larvae can occur in many temperate latitudes, where these ecologically and commercially important fish are known to spawn, and may contribute to the high mortality of cod embryos and larvae in their natural environment.

The superolateral approach for aspiration of total hip arthroplasties.

Hip aspiration for exclusion of infection is a common procedure prior to revision arthroplasty. Some patients with total hip arthroplasties develop superomedial migration of the prosthesis, while others can have extensive heterotopic bone formation. The traditional approaches may be contraindicated due to overlying bowel or bone anterior/lateral to the prosthesis. These patients are candidates for the superolateral approach. With the patient supine marks are made on the skin at (i) the greater trochanter and (ii) a superior site that aligns with the neck of the prosthesis and soft-tissue window. The needle enters the lateral skin parallel to the table top. Usually only one pass is necessary.

Limited magnetic resonance imaging in the diagnosis of occult hip fractures.

The majority of fractures of the hip are readily diagnosed on the basis of the clinical findings and plain radiographs. When initial X-rays are negative or equivocal and a high clinical suspicion exists for an occult hip fracture, additional diagnostic studies need to be performed. Use of limited magnetic resonance imaging in the diagnosis of occult hip fracture upon initial presentation to the emergency department is reported.

Preoperative physical therapy in primary total knee arthroplasty.

In order to evaluate the efficacy of preoperative physical therapy for patients undergoing elective primary total knee arthroplasty, 10 patients completed 6 weeks of physical therapy before surgery (PT group). Ten patients served as controls (C group). Subjects were tested at baseline (PT only), before surgery, 6 weeks after surgery, and 3 months after surgery using the Hospital for Special Surgery knee rating scale, range of motion, thigh circumference, walking speed, Cybex II isokinetic knee flexion, and extension testing, and computed tomography scanning for cross-sectional muscle area. Hospital stay and need for physical therapy after inpatient rehabilitation were also compared. Physical therapy produced modest gains in isokinetic flexion strength in these severely arthritic knees but no difference in extension strength. The decrease in isokinetic strength after surgery was not affected by preoperative physical therapy. Muscle area did not decrease significantly for the PT group, but it did decrease for the C group after surgery. While postoperative strength differences could not be demonstrated, preoperative physical therapy preserved thigh muscle area after surgery. The clinical significance of this finding is uncertain. Consequently, this study failed to support the routine use of preoperative physical therapy in knee replacement surgery.

A pseudo-outbreak of respiratory infection with Acinetobacter species.

We describe an apparent outbreak of respiratory infection with Acinetobacter species involving 14 patients over 8 days. Epidemiological investigation revealed two consecutive pseudo-outbreaks of infection caused by two consecutive, unrelated laboratory errors in the processing of sputum, nasopharyngeal, and endotracheal aspirates.

Characterization of gene expression of a p53 homologue in the soft-shell clam (Mya arenaria).

Expression of a clam p53 homologue was examined in tissues of the soft-shell clam, Mya arenaria, from Beal's Island, Maine. Southern analysis reveals that p53, in this population, is a single copy gene. A 1.7 to 1.9-kb p53 mRNA was detected at very low levels in normal adult gonadal tissue. This transcript is similar in size to that of vertebrate p53 genes. RNAs were harvested from several tissues, including individual clam gonads during gametogenesis. These were hybridized in ribonuclease (RNase) protection assays to a p53 antisense probe designed from the clam p53 cDNA sequence. RNase protection profiles indicate that p53 mRNA is expressed in adductor muscle, gill, and gonads of both sexes. Although p53 mRNA is expressed throughout gametogenesis in mature male and female gonads, ovaries have significantly higher levels of expression. The significance of our findings to the study of normal clam gametogenesis and to etiology of gonadal tumors is discussed.

Imaging of skeletal and soft tissue injuries in and around the knee.

This article addresses the place of MR imaging in the diagnosis of knee injuries. MR imaging of subtle fractures and bone contusions is described with emphasis placed on those types of fractures frequently associated with specific patterns of soft tissue injury. Soft tissue injuries are divided into ligamentous, meniscal, and muscular; each is discussed separately. The MR imaging appearance of these lesions is explained.

Detection of mutant p53 in clam leukemia cells.

Leukemia in the soft-shell clam, Mya arenaria, is characterized by tumor cells which are detected initially in the hemolymph. This disease is much more common in clams inhabiting polluted waters, suggesting an environmental component to its pathogenesis. In this study, leukemia cells were identified using a murine monoclonal antibody, 1E10, which recognizes a leukemia-specific protein expressed by tumor cells. Mutant p53 protein was detected using a murine monoclonal antibody (PAb 240) which reacts with mutant p53. Using immunofluorescence, the reactivity of clam cells to the 1E10 antibody was evaluated along with mutant p53 protein reactivity. Reverse transcriptase-polymerase chain reactions followed by sequence analyses were utilized to examine clams with hemocytes reacting with the p53 antibody for possible p53 gene mutations. Mutant p53 protein was expressed by tumor cells from five animals with advanced disease (in which greater than 90% of cells reacted with 1E10). A C-->G transversion was detected at the end of exon 6 from two of the five animals that reacted with both the mutant p53 antibody and 1E10. This substitution changes the amino acid of this codon from proline to alanine. Overall, our results suggest that environmentally induced alterations in p53 can contribute to the pathogenesis of leukemia in soft-shell clams inhabiting polluted water and/or sediment.

Acanthamoeba: ecology, pathogenicity and laboratory detection.

Acanthamoeba spp. are ubiquitous free-living protozoa found in a wide range of environmental niches. They are resistant to disinfectants, temperature variation and desiccation and are responsible for two recognised diseases in humans, granulomatous amoebic encephalitis and keratitis. Both infections are rare, although the latter is currently receiving more attention following the association between Acanthamoeba and the wearing of contact lenses. Laboratory diagnosis is unusual but not beyond the bounds of most routine clinical microbiology departments. In this review the various aspects surrounding the ecology, pathogenicity and laboratory detection of Acanthamoeba spp. are considered.