Statistical models of tumour onset and growth for modern breast cancer screening cohorts.

Historically, multi-state Markov models have been used to study breast cancer incidence and mammography screening effectiveness. In recent years, more biologically motivated continuous tumour growth models have emerged as alternatives. However, a number of challenges remain for these models to make use of the wealth of information available in large mammography cohort data. In particular, methodology is needed to address random left truncation and individual, asynchronous screening. We present a comprehensive continuous random effects model for the natural history of breast cancer. It models the unobservable processes of tumour onset, tumour growth, screening sensitivity, and symptomatic detection. We show how the unknown model parameter values can be jointly estimated using a prospective cohort with diagnostic data on age and tumour size at diagnosis, and individual screening histories. We also present a microsimulation study calibrated to population breast cancer incidence data, and to data on mode of detection and tumour size. We highlight the importance of adjusting for random left truncation, derive tumour doubling time distributions for screen-detected and interval cancers, and present results concerning the relationship between tumour presence time and age at diagnosis.

Early onset of inflammation during ontogeny of bipolar disorder: the NLRP2 inflammasome gene distinctly differentiates between patients and healthy controls in the transition between iPS cell and neural stem cell stages.

Neuro-inflammation and neuronal communication are considered as mis-regulated processes in the aetiology and pathology of bipolar disorder (BD). Which and when specific signal pathways become abnormal during the ontogeny of bipolar disorder patients is unknown. To address this question, we applied induced pluripotent stem cell (iPSC) technology followed by cortical neural differentiation on adipocyte-derived cells from BD type I patients (with psychotic episodes in psychiatric history) and healthy volunteers (controls). RNA sequencing in iPSC and cortical neural stem cell (NSC) lines were used to examine alterations between the transcriptomes from BD I and control samples during transition from the pluripotent stage towards the neural developmental stage. At the iPSC stage, the most highly significant differentially expressed gene (DEG) was the NLRP2 inflammasome (P=2.66 × 10-10). Also among 42 DEGs at the NSC stage, NLRP2 showed the strongest statistical significance (P=3.07 × 10-19). In addition, we have also identified several cytoskeleton-associated genes as DEGs from the NSC stage, such as TMP2, TAGLN and ACTA2; the former two genes are recognised for the first time to be associated with BD. Our results also suggest that iPSC-derived BD-cortical NSCs carry several abnormalities in dopamine and GABA receptor canonical pathways, underlining that our in vitro BD model reflects pathology in the central nervous system. This would indicate that mis-regulated gene expression of inflammatory, neurotransmitter and cytoskeletal signalling occurs during early fetal brain development of BD I patients.

Improving environmental and biodiversity monitoring in the Baltic Sea using DNA barcoding of Chironomidae (Diptera).

As for many other regions, environmental and biodiversity monitoring of the brackish Baltic Sea suffers from low species resolution for several taxa. One such case is the benthic larvae of midges Chironomidae (Diptera), which are estimated to constitute about 30% of the macrozoobenthos species of the Baltic Sea and are important indicators of environmental quality. We assessed the usefulness of COI (cytochrome oxidase I) gene barcoding to improve species resolution and its potential for implementation in monitoring programmes. Neighbour-Joining, Maximum parsimony and Bayesian-inference analyses all provided high congruency with morphological analyses of adult males for almost all 42 species studied. Barcoding was helpful to elucidate some cases of taxonomical difficulties, such as synonyms. In contrast to the high identification accuracy when using our local database, there were a number of cases where matching with GenBank and BOLD provided puzzling results. For reliable species identification at least 15-30 specimens from 5-10 well-distributed sites within the geographical range of the species might be needed in a database to adequately cover the intraspecific variability of chironomids. Implementation of DNA barcoding, as applied here, in monitoring would result in an increase from at present less than 10% to more than 90% successful chironomid species identification of Baltic Sea benthic samples, as it also would for many nearby lakes. Routine monitoring of benthic environmental samples based on Next-Generation sequencing techniques would provide a cost effective way to obtain a taxonomically much more complete assessment of environmental quality and biodiversity, as required by EU directives and national legislation.

Critical and complex role of N-methyl-D-aspartate receptors in long-term depression at CA3-CA1 synapses in the developing hippocampus.

In the developing brain synaptogenesis is accompanied by elimination of synapses possibly initiated by activity-dependent synaptic plasticity such as long-term depression (LTD). Experiments using 2nd postnatal week hippocampal CA3-CA1 synapses have previously shown that these synapses, in contrast to those in the more adult brain, are easily depressed even during very low frequency (0.05-0.2 Hz) activity. We have now addressed the question whether such stimulation actually results in LTD, and if so, under which conditions this occurs. By introducing 30-60 min of stimulus interruption following 900 stimuli at 0.2 Hz and 0.05 Hz we found this stimulation to result in an LTD of -37% and -24%, respectively. The LTD following 0.2 Hz stimulation did not differ significantly from that resulting from 900 stimuli using the common LTD-inducing frequency of 1 Hz. When 0.2 Hz and 1 Hz stimulations were applied in the presence of a combined N-methyl-d-aspartate receptor (NMDAR)/mGluR blockade the LTDs were only marginally smaller. However, the LTD observed under this latter condition was labile in that it reversed (de-depressed) by spontaneous and/or ambient NMDAR activity (labile LTD). 0.2 and 1 Hz-evoked NMDAR activity resulted in LTD not de-depressed by spontaneous and/or ambient NMDAR activity (stable LTD) and in little or no labile LTD. The stable LTD was fully de-depressed by high frequency-evoked NMDAR activity. 0.2 and 1 Hz-evoked mGluR activity impaired the labile LTD but did not result in stable LTD. In conclusion, in 2nd postnatal week CA3-CA1 synapses LTD is induced at frequencies well below one Hz as well as in the absence of NMDAR activity. Very low/low frequency-evoked NMDAR activity stabilizes LTD by raising its threshold for NMDAR-dependent de-depression. LTD at these developing synapses thus seems adapted for ease of induction as well as of de-depression.

In vivo mammary tumourigenesis in the Sprague-Dawley rat and microdosimetric correlates.

Standard methods for risk assessments resulting from human exposures to mixed radiation fields in Space consisting of different particle types and energies rely upon quality factors. These are generally defined as a function of linear energy transfer (LET) and are assumed to be proportional to the risk. In this approach, it is further assumed that the risks for single exposures from each of the radiation types add linearly. Although risks of cancer from acute exposures to photon radiations have been measured in humans, quality factors for protons and ions of heavier atomic mass are generally inferred from animal and/or cellular data. Because only a small amount of data exists for such particles, this group has been examining tumourigenesis initiated by energetic protons and iron ions. In this study, 741 female Sprague-Dawley rats were irradiated or sham irradiated at approximately 60 days of age with 250 MeV protons, 1 GeV/nucleon iron ions or both protons and iron ions. The results suggest that the risk of mammary tumours in the rats sequentially irradiated with 1 GeV/nucleon 56Fe ions and 250 MeV protons is less than additive. These data in conjunction with earlier results further suggest that risk assessments in terms of only mean LETs of the primary cosmic rays may be insufficient to accurately evaluate the relative risks of each type of particle in a radiation field of mixed radiation qualities.

Survival and Persistence of Alternaria dauci in Carrot Cropping Systems.

Alternaria dauci was recovered in California from carrot crop residue and from volunteer carrot plants in fallow carrot fields. The fungus was not recovered from common weeds surrounding fallow fields. To evaluate further the survival of A. dauci on carrot crop residue, infected carrot leaf tissue was placed in fields or in soil in greenhouse pots, and recovered over time. In California, A. dauci was recovered from infected leaf tissue in both fallow and irrigated fields for as long as 1 year. In Florida, A. dauci was recovered from infected leaf tissue in fallow fields for up to 30 weeks. In greenhouse experiments, A. dauci was recovered from infected leaf tissue for as long as 1 year in dry soil, but only up to 30 weeks in soil that was watered weekly. To determine the infectivity of A. dauci borne on carrot crop residue, infected carrot crops were incorporated into organic and mineral field soils, and soil samples were collected over time. Carrot seed were planted in collected soil, and seedling infection by A. dauci was recorded. Seedling infection was detected up to 13 and 14 weeks after crop incorporation in organic and mineral soil, respectively. Seedling infection was detected for up to 5 weeks in soil that remained dry compared with 3 weeks in flooded soil.

Photodynamic therapy in the canine prostate using motexafin lutetium.

Our purpose was to determine the feasibility of comprehensive treatment of the canine prostate with photodynamic therapy (PDT) using motexafin lutetium (Lu-Tex) and to evaluate the toxicity and tissue effects associated with this treatment. Twenty-five adult male beagles with normal prostate glands were given an i.v. injection of the second-generation photosensitizer Lu-Tex (2-6 mg/kg). An additional two dogs were used as controls and did not receive any photosensitizing drug. All 27 dogs underwent laparotomy to expose the prostate. Three hours postinjection, a total dose of 75-150 J/cm of 732 nm laser light was delivered interstitially and/or transurethrally to the prostate via cylindrical diffusing fibers. Dogs were euthanized between 2 days and 3 months after PDT. All subjects were monitored for clinical evidence of toxicity. Specimens were examined macroscopically and microscopically to characterize the tissue reaction and assess extent of tissue effect as a result of treatment. Interstitial and/or transurethral PDT were successfully delivered in all dogs with no perioperative complications. No clinical evidence of acute urinary obstruction or rectal bleeding was noted. At all dose levels, macroscopic and microscopic evaluation revealed a prostatic tissue reaction characterized initially (within 48 h) by inflammation and necrosis followed by fibrosis and glandular epithelial atrophy. Comprehensive treatment of the entire prostate could be achieved using the interstitial alone approach or combined transurethral and interstitial approach. The transurethral alone approach did not result in complete coverage of the prostate. Dogs receiving transurethral or combined interstitial and transurethral treatment developed erythema and urethral epithelial disruption at all dose levels. Those receiving combined treatment at the highest dose level (Lu-Tex 6 mg/kg, 150 J/cm light) developed urethral fistulae and peritonitis. Dogs treated with the interstitial alone approach were found to have the least amount of urethral damage. Comprehensive treatment of the canine prostate with Lu-Tex PDT is feasible using an interstitial alone or combined interstitial and transurethral approach. The interstitial alone technique results in the least amount of toxicity. The prostatic tissue reaction to treatment is characterized by initial inflammation and necrosis followed by fibrosis and glandular epithelial atrophy.

A New Disease of Flowering Dogwood Caused by Colletotrichum acutatum.

In 1992 symptoms consisting of twig dieback, canker-like deformities, and often, mortality of all sizes of container-grown flowering dogwood trees (Cornus florida L.) was frequently observed in northern Florida where nursery production of dogwood is concentrated. Twigs, branches, growing points, and portions of main trunks up to 3 cm in diameter were killed. On affected portions, leaves drooped downward but did not wilt severely; they later became silvery, gray-green and eventually died but did not drop. Affected trees produced flower buds, but the flower buds did not open. Many trees died and others were destroyed because of unsightly dead limbs and deformed trunks. The inciting pathogen was not satisfactorily determined. After 1994, disease incidence subsided, but in 1999, it reappeared and caused serious economic damage. In 1999, we followed the disease from propagation through several stages of nursery production, including growth in large containers. A Colletotrichum species was consistently isolated from acervuli produced in abundance near the soil line on diseased, rooted cuttings and small plants. The same fungus was consistently isolated from acervuli produced on diseased leaves and twigs of larger plants and small trees. Conidia of the isolated pathogen were used to inoculate small dogwood trees kept at 100% RH for 2 days then grown in a greenhouse. Within 23 weeks, a slowly developing leaf spot was produced on inoculated plants. After 3 months, numerous acervuli were produced on inoculated leaves and on adjacent small twigs from which the same pathogen was consistently reisolated. The symptoms and pathogen were not consistent with descriptions of anthracnose incited by Discula destructiva. On potato-dextrose agar (PDA), spore masses were bright red-orange on lawn plates inoculated with large numbers of conidia. Conidiomata did not produce setae in culture nor did acervuli on diseased plant material. On PDA, mycelial growth was tufted and pale-gray. The reverse side of colonies was buff to cream-colored or pale-gray to tan but never dark. With age, a pale pink or orange-pink pigment often formed within the agar media (1,3). When produced on PDA, most conidia were elliptical and elongated with abruptly tapering ends. An average of 18% had both ends slightly rounded; none had only one end abruptly tapered or rounded. Conidia measured 15.3 × 4.78 µm; the length/width ratio was 3.20 (1,3). The teleomorph was not found on diseased plant material or in culture. Analysis of DNA by polymerase chain reaction (PCR) using the CaInt2-ITS4 and CgInt-ITS4 primer pairs (1,2) and by comparison of PCR products with those produced by C. acutatum pathotypes isolated from leatherleaf fern, lime, post-bloom fruit drop-affected Navel orange fruit, and strawberry fruit, and with isolates of C. gleosporiodes from citrus, Camellia, Nandina, holly, and strawberry indicated that the fungus was C. acutatum. The amplified PCR product (approximately 490 bp) obtained from all dogwood isolates using primers CaInt2 and ITS4 was consistent with the size of product expected from C. acutatum. No products were produced with the CgInt-ITS4 primer pair. Based on the morphology of conidia, growth in culture and PCR results, the pathogen was identified as C. acutatum and represents the first report of this pathogen on flowering dogwood. References: (1) J. E. Adaskaveg and R. J. Hartin. Phytopathology 87:979, 1997. (2) A. E. Brown et al. Phytopathology 86:523, 1996. (3) B. C. Sutton. Pages 1-27 in: Colletotrichum: Biology, pathology and control. Brit. Soc. For Plant Pathol., CAB International, 1992.

National Institutes of Health--supported opportunities in Comparative Medicine.

The Comparative Medicine (CM) area of the National Institutes of Health (NIH) is a major source of support for research on laboratory animals, training of laboratory animal specialists, and support of shared, regional animal resources. We present a brief history of CM at NIH and the major mechanisms by which it accomplishes its goals in programs located across the United States.

Cestodiasis with intestinal diverticulosis in a lesser flamingo (Phoeniconaias minor).

An adult female lesser flamingo (Phoeniconaias minor), caught in the African Rift Valley in 1991 and subsequently housed at the Baltimore Zoo, died of severe visceral gout in 1996. Necropsy revealed a white, moderately firm, nodular lesion, 1 cm in diameter, in the serosal wall of the small intestine. Although it was initially thought to be a tumor or focal granuloma, histologic examination revealed multiple cestodes deeply embedded at the base of the crypts between the intestinal villi, with their massive scolices (up to 3.4 mm in diameter) distending these spaces into multiple diverticulae. The mucosal epithelium surrounding the scolices was severely attenuated. Around the diverticulae, in the submucosa and muscularis, was a mild to moderate lymphocytic reaction and mild fibrosis. The proximity of multiple scolices and extensive invasion of host tissue suggested that the infection occupied a preexisting lesion. The cestodes were cyclophyllids but were distinct from any species previously reported from flamingos. Helminths should be included in differential diagnoses for gastrointestinal nodules in flamingos.

Nontraumatic osteonecrosis: MR perfusion imaging evaluation in an experimental model.

RATIONALE AND OBJECTIVES: Because the nature and time course of changes in early, nontraumatic osteonecrosis at perfusion and magnetic resonance (MR) imaging are unknown, the authors evaluated this technique in the assessment of early osteonecrosis with a nontraumatic model. MATERIALS AND METHODS: Five rabbits underwent intravenous injection of lipopolysaccharide endotoxin followed by intramuscular injection of methylprednisolone. MR imaging of the femora was performed before and at weekly intervals after endotoxin injection. Histologic findings from the areas of osteonecrosis were correlated with the findings of MR imaging and MR perfusion studies. RESULTS: Histologic evaluation showed osteonecrosis in six femora of four animals 2-4 weeks after endotoxin injection. Findings on T1-weighted images of the femur were normal in all animals; T2-weighted images of one femur showed equivocal changes. On MR perfusion images, the baseline mean peak percentage of enhancement was 52.7% +/- 12.6. In the six areas without osteonecrosis, the mean percentage of enhancement was similar to the baseline percentage of enhancement at 1 week (62.2% +/- 31.2). In the four areas with diffuse osteonecrosis, there was essentially no contrast enhancement 1-4 weeks after endotoxin injection. CONCLUSION: T1- and T2-weighted MR imaging is insensitive to the presence of early nontraumatic osteonecrosis. MR perfusion imaging might be useful to detect early nontraumatic osteonecrosis.

Cholesterol granulomas in mice deficient in apolipoprotein E.

Three 12- to 16-month-old female B6,129 apolipoprotein E-deficient mice were presented for necropsy because of bilateral masses in the gluteal region. Histopathologic examination revealed cholesterol granulomas extending from the superficial dermis to the underlying gluteal muscles. Microscopic granulomas were found on the face, perivaginal tissues, mesenteric lymph nodes, spleen, joints, kidneys, and choroid plexus. Other lesions included severe atherosclerosis of the aortic valves, aorta, pulmonary artery, and renal artery. Here we show that macroscopic cholesterol granulomas can develop in apolipoprotein E-deficient that receive a normal rodent diet.

Processing postmortem specimens with C18-carboxypropylbetaine and analysis by PCR to develop an antemortem test for Mycobacterium avium infections in ducks.

Mycobacterium avium is the causative agent of the avian mycobacteriosis commonly known as avian tuberculosis (ATB). This infection causes disseminated disease, is difficult to diagnose, and is of serious concern because it causes significant mortality in birds. A new method was developed for processing specimens for an antemortem screening test for ATB. This novel method uses the zwitterionic detergent C18-carboxypropylbetaine (CB-18). Blood, bone marrow, bursa, and fecal specimens from 28 ducks and swabs of 20 lesions were processed with CB-18 for analysis by smear, culture, and polymerase chain reaction (PCR). Postmortem examination confirmed nine of these birds as either positive or highly suspect for disseminated disease. The sensitivities of smear, culture, and PCR, relative to postmortem analysis and independent of specimen type, were 44.4%, 88.9%, and 100%, respectively, and the specificities were 84.2%, 57.9%, and 15.8%, respectively. Reductions in specificity were due primarily to results among fecal specimens. However, these results were clustered among a subset of birds, suggesting that these tests actually identified birds in early stages of the disease. Restriction fragment length polymorphism mapping identified one strain of M. avium (serotype 1) that was isolated from lesions, bursa, bone marrow, blood, and feces of all but three of the culture-positive birds. In birds with confirmed disease, blood had the lowest sensitivity and the highest specificity by all diagnostic methods. Swabs of lesions provided the highest sensitivity by smear and culture (33.3% and 77.8%, respectively), whereas fecal specimens had the highest sensitivity by PCR (77.8%). The results of this study indicate that processing fecal specimens with CB-18, followed by PCR analysis, may provide a valuable first step for monitoring the presence of ATB in birds.

Feline asthma syndrome in African lions (Panthera leo).

Feline asthma syndrome, previously recognized only in domestic cats, was diagnosed in three captive African lions (Panthera leo), one of which died as a result of the condition. Two of the lions displayed progressive signs for 7 yr, including severe bouts of coughing, wheezing, dyspnea, rhonchi, and tachypnea that were most severe during the spring and summer, and the third lion displayed acute signs only once. Scattered to diffuse increased interstitial markings, peribronchial cuffing, and focal atelectasis were visible in radiographs. At necropsy, multiple subpleural bullae, 2-3 cm in diameter, were scattered throughout the lung tissue. There were thick-walled bronchi and bronchioles filled with thick grayish mucus, and alveolar spaces were enlarged with severe, diffuse, banded multifocal areas of alveolar wall fibrosis. The lions had significantly elevated IgE type I immediate hypersensitivity responses to recognized aeroallergens. The captive management of lions should address the design and maintenance of allergen-free air supplies. Ventilation systems should be examined routinely and thoroughly cleaned of any residue. The frequency of examination should increase during the summer. Lions and other large cats should be routinely screened for IgE aeroallergen-specific titers, asthma cases should be treated promptly with prednisolone, and investigations of etiology should be initiated.

Comparison of acute endotoxin-induced lesions in A/J and C57BL/6J mice.

Resistance to the action of endotoxin varies among inbred strains of mice, indicating that a component of this resistance has a genetic basis. Different responses to endotoxin that are characteristic of individual inbred strains represent phenotypes that can be used to genetically map the response modifier genes. This study compares the acute histologic lesions in 8-week-old male A/J and C57BL/6J (B6) mice injected intraperitoneally with endotoxin of E. coli O265:B6 (15 mg/kg). Animals of both strains exhibited splenitis, splenic lymphoid hyperplasia, splenic lymphoid necrosis, and sequestration of neutrophils in the pulmonary alveoli. The B6 mice showed increased margination of white blood cells to the pulmonary vascular endothelium relative to A/J mice. A large number of degenerating neutrophils was observed in the liver sinusoids of most B6 animals, while this lesion was much less severe in A/J mice. This difference was quantified, demonstrating a highly significant difference in neutrophil infiltration in B6 mice relative to A/J mice. Analysis of this phenotype in F1 mice demonstrates that major genes encoding the trait are not X-linked, imprinted, or maternally inherited and do not show the codominant inheritance expected if Lps(d) were primarily responsible. The distinctive, quantitative nature of these differences provides a useful assay for mapping genes that modify endotoxin responsiveness using the AXB and BXA recombinant inbred (RI) strains derived from A/J and B6 mice.

Real-time bronchoscope tip localization enables three-dimensional CT image guidance for transbronchial needle aspiration in swine.

STUDY OBJECTIVE: To determine the feasibility of using real-time bronchoscope position technology coupled with previously acquired three-dimensional CT data to enhance transbronchial needle aspiration (TBNA). DESIGN: Eight swine were given percutaneously created target lesions for TBNA. A miniature position sensor was placed at the tip of a bronchoscope, and real-time position information during bronchoscopy was presented on a monitor simultaneously displaying previously acquired three-dimensional CT data. INTERVENTIONS: TBNA of target lesions and submucosal ink-spot injection of computer-generated targets. MEASUREMENTS AND RESULTS: TBNA specimens revealed successful aspiration of target material. Distances between ink marks made at computer-generated tracheal targets varied, on average (+/- SD), 4.2 mm +/- 2.6 mm from predetermined computer-distance coordinates. CONCLUSION: Real-time bronchoscope position technology coupled with previously acquired CT images may aid with TBNA of nonvisible extrabronchial lesions.

Gongylonema macrogubernaculum in captive African squirrels (Funisciurus substriatus and Xerus erythropus) and lion-tailed macaques (Macaca silenus).

Necropsies performed between 1989 and 1995 on 15 African rope squirrels (Funisciurus substriatus) and 20 African ground squirrels (Xerus erythropus) from the Baltimore Zoo revealed 13 cases of gongylonemiasis. Nematodes were embedded in the epithelium of the esophagus, pharynx, buccal mucosa, and tongue, resulting in varying degrees of esophagitis, pharyngitis, stomatitis, and glossitis, respectively. Routine fecal examinations were negative, and the nematodes appeared to be unaffected by repeated treatments with ivermectin. Most of the affected animals had shown clinical signs of dyspnea and/or inanition and emaciation. Suppurative rhinitis was also a frequent finding at necropsy and was associated with the presence of the nematodes in eight animals. Dissection of whole nematodes from formalin-fixed specimens revealed morphologic features consistent with Gongylonema macrogubernaculum, a species previously only reported in nonhuman primates. The squirrels were housed in the same building with numerous primate species, and a review of pathology records revealed esophageal gongylonemiasis in three lion-tailed macaques (Macaca silenus), lingual gongylonemiasis in a spotnose monkey (Cercopithecus buettikoferi), and buccal gongylonemiasis in a brown-headed tamarin (Saguinus fuscicollis). Examination of whole nematodes dissected from one of the lion-tailed macaques also demonstrated the unique morphology of G. macrogubernaculum. Nematodes belonging to the species Gongylonema are acquired by ingestion of the intermediate host, the cockroach. This is the first report of G. macrogubernaculum in a nonprimate species and suggests that captive African squirrels can serve as reservoir hosts for this parasite in a zoo environment.

Asymmetric infectivity of pseudorecombinants of cabbage leaf curl virus and squash leaf curl virus: implications for bipartite geminivirus evolution and movement.

The bipartite geminiviruses squash leaf curl virus (SqLCV) and cabbage leaf curl virus (CLCV) have distinct host ranges. SqLCV infects a broad range of plants within the Cucurbitaceae, including pumpkin and squash, and CLCV has a broad host range within Brassicaceae that includes cabbage and Arabidopsis thaliana. Despite this, the genomic A components of these viruses share a high degree of sequence identity, particularly in the gene encoding the replication protein AL1, and their common regions are 77% identical. However, there is unexpected sequence diversity in the common regions of the two CLCV genomic A and B components, these being only 80% identical. Based on these sequence similarities, we investigated the host range properties of pseudorecombinants of SqLCV and CLCV. We found that in a pseudorecombinant virus consisting of the A component of CLCV and the B component of SqLCV, both components replicated in tobacco protoplasts, and this pseudorecombinant was infectious and caused systemic disease in Nicotiana benthamiana, a common host to all bipartite geminiviruses. However, this pseudorecombinant did not move systemically in pumpkin or Arabidopsis, despite the demonstrated replication compatibility of the genome components. As a result of the greater sequence differences between the common regions, the pseudorecombinant of SqLCV A and CLCV B components neither replicated the CLCV B component nor systemically infected any of the hosts tested. These findings demonstrate that for different geminiviruses with distinct host ranges, the replication origins and AL1 proteins can be sufficiently similar to permit infectious pseudorecombinants, but replication alone is not sufficient to cause systemic disease, and host range may ultimately be limited at the level of movement. The results of this study further suggest that CLCV is an evolving virus that can provide insights into how new bipartite geminiviruses arise from mixed infections.

Age-dependent and lobe-specific spontaneous hyperplasia in the brown Norway rat prostate.

We showed previously that exogenously administered testosterone caused age- and lobe-specific overgrowth of the prostate in Brown Norway rats. A common feature observed in testosterone-treated animals was cell hypertrophy in each of the ventral, dorsal, and lateral lobes of both young (6 mo old) and old (24 mo old) rats. By contrast, hyperplasia was seen only in the dorsal and lateral lobes of old rats treated with testosterone. These observations prompted us to examine whether age- and lobe-specific overgrowth might also occur in untreated rats as a consequence of the endogenous hormonal milieu. To this end, blood and prostates were collected from a large number (25-30 rats per group) of 4- to 6-mo-old (young) and 21- to 24-mo-old (old) Brown Norway rats. Both serum testosterone (-45%) and estradiol (-22%) concentrations decreased significantly with age, but the greater magnitude of the decrement in testosterone relative to estradiol led to a reduction in the serum testosterone:estradiol ratio. Paradoxically, although the prostate is androgen dependent, the wet weight, protein, and DNA contents increased significantly with age in the dorsal and lateral lobes of old rats despite the decrease in testosterone level. Histologic examination revealed that the increased weights and DNA contents of the dorsal and lateral lobes in old rats coincided with an increased number of epithelial cells in the distal and intermediate segments of these lobes, indicative of hyperplasia but independent of change in cell size. Taken together, these results show a spontaneous age-related overgrowth of cells in the dorsal and lateral prostatic lobes of old Brown Norway rats despite diminished serum testosterone concentrations. The aging Brown Norway rat, therefore, may be a useful model for studies of some aspects of the pathogenesis underlying spontaneous age-related prostatic hyperplasia.