Immunohistochemical Expression of Potential Therapeutic Targets in Canine Thyroid Carcinoma

Background

Thyroid carcinoma is a common endocrine tumor in the dog. Local invasive growth frequently precludes surgical excision and, in up to 38% of dogs, the tumor has already metastasized by the time of diagnosis. Therefore, it is important to investigate new treatment modalities that may be useful for the large number of dogs with inoperable tumors or metastatic disease.

Hypothesis/Objectives

To investigate the immunohistochemical expression of potential therapeutic targets in canine thyroid tumors.

Animals

74 dogs with thyroid neoplasia.

Methods

Immunohistochemistry was performed for thyroglobulin, calcitonin, vascular endothelial growth factor (VEGF), p53, cycloxygenase‐2 (cox‐2), and P‐glycoprotein (P‐gp).

Results

Fifty‐four (73%) tumors were classified as follicular cell thyroid carcinomas (FTCs) and 20 (27%) as medullary thyroid carcinomas (MTCs). Eighty percent of FTCs and all MTCs had a high percentage (76–100%) of neoplastic cells immunopositive for VEGF. Thirteen percent of FTCs and 50% of MTCs expressed cox‐2. Seven percent of FTCs and 70% of MTCs expressed P‐gp. No tumor was immunopositive for p53 expression. Expression of VEGF (P = .034), cox‐2 (P = .013), and P‐gp (P < .001) was significantly higher in MTCs compared to FTCs.

Conclusions and Clinical Importance

VEGF is a potential therapeutic target in both FTC and MTC in dogs. Cox‐2 and P‐gp may be useful molecular targets in canine MTC.     

Molecular phylogeny and morphological characterization of the aetiological agent of sour rot on fruits and vegetables in Brazil

In Brazil, sour rot is an important postharvest disease on fruits and vegetables. Geotrichum candidum (synonym Galactomyces candidus) has been reported as the main species causing this disease. However, the identity of the causal agent is still uncertain. This research aimed to determine the identity of 165 fungal isolates associated with sour rot obtained from fruits and vegetables in Brazil, and to evaluate the effect of different temperatures on the incidence of sour rot on artificially inoculated tomato fruits. Based on the phylogenetic analyses of DNA sequences from the D1/D2 domain of the large subunit (LSU) rRNA gene and morphological analyses, 129 samples belonged to Galactomyces candidus, 15 to G. candidum var. citri-aurantii, 6 to G. phurueaense, 2 to Gal. pseudocandidus, 1 to Hyphopichia burtonii, 1 to H. khmerensis, 3 to Saccharomycopsis crataegensis, 1 to S. vini, 1 to Magnusiomyces tetrasperma, 1 to Trichosporon coremiiforme, and 1 to Zygoascus meyerae. Two new species were found, namely, Geotrichum solani (on potato) and Geotrichum spondiadis (on red mombin). All isolates were pathogenic when inoculated on healthy tomato fruits, including the new species of Geotrichum, which were also inoculated into their respective hosts, that is, potatoes and red mombins. To the best of our knowledge, this study is the first to report the presence of five other genera besides Geotrichum associated with sour rot on fruits and vegetables in Brazil, which demonstrates the diversity of fungi and yeasts associated with this disease.     

Inheritance of the resistant reaction of the lettuce cultivar `Grand Rapids' to the southern root-knot nematode Meloidogyne incognita (Kofoid & White) Chitwood

Resistance to the southern root-knot nematode Meloidogyne incognita Chitwood would be an important attribute of lettuce Lactuca sativa L. cultivars adapted to both protected and field cultivation in tropical regions. `Grand Rapids' and a few other cultivars are reported to be resistant to this nematode. In this paper, we studied the inheritance of the resistant reaction of `Grand Rapids' (P₂) in a cross with a standard nematode-susceptible cultivar Regma-71 (P₁). F₁(Regina-71 × Grand Rapids) and F₂ seed were obtained, and inoculated along with the parental cultivars with different races of M. incognita to evaluate nematode resistance. Broad sense heritability estimates for the number of galls and of egg masses per root system, gall size and gall index were generally in the order of 0.5 or higher. Class distributions of these variables over generations P₁, P₂, F₁ and F₂ were in agreement with simulated theoretical distributions based on monogenic inheritance models. F₃ families were obtained from randomly sampled F₂ plants and tested for reaction to the nematode. The frequency ratio of homozygous resistant, segregating and homozygous susceptible F₃ families did not differ from the 1:2:1 ratio expected from monogenic inheritance. M. incognita resistance appears to be under control of a single gene locus. The Grand Rapids allele (for which the symbol Me is proposed) is responsible for the resistant reaction, and shows high (though incomplete) penetrance, variable expressivity and predominantly additive gene action. This revised version was published online in July 2006 with corrections to the Cover Date.     

Convergence of developmental mutants into a single tomato model system: 'Micro-Tom' as an effective toolkit for plant development research

Background

The tomato (Solanum lycopersicum L.) plant is both an economically important food crop and an ideal dicot model to investigate various physiological phenomena not possible in Arabidopsis thaliana. Due to the great diversity of tomato cultivars used by the research community, it is often difficult to reliably compare phenotypes. The lack of tomato developmental mutants in a single genetic background prevents the stacking of mutations to facilitate analysis of double and multiple mutants, often required for elucidating developmental pathways.

Results

We took advantage of the small size and rapid life cycle of the tomato cultivar Micro-Tom (MT) to create near-isogenic lines (NILs) by introgressing a suite of hormonal and photomorphogenetic mutations (altered sensitivity or endogenous levels of auxin, ethylene, abscisic acid, gibberellin, brassinosteroid, and light response) into this genetic background. To demonstrate the usefulness of this collection, we compared developmental traits between the produced NILs. All expected mutant phenotypes were expressed in the NILs. We also created NILs harboring the wild type alleles for dwarf, self-pruning and uniform fruit, which are mutations characteristic of MT. This amplified both the applications of the mutant collection presented here and of MT as a genetic model system.

Conclusions

The community resource presented here is a useful toolkit for plant research, particularly for future studies in plant development, which will require the simultaneous observation of the effect of various hormones, signaling pathways and crosstalk.     

Agarose gel electrophoresis of cerebrospinal fluid proteins of dogs after sample concentration using a membrane microconcentrator technique

BACKGROUND: Cerebrospinal fluid (CSF) is produced in the cerebral ventricles through ultrafiltration of plasma and active transport mechanisms. Evaluation of proteins in CSF may provide important information about the production of immunoglobulins within the central nervous system as well as possible disturbances in the blood-brain barrier. OBJECTIVE: The objective of this study was to measure the concentration and fractions of protein in CSF samples using a membrane microconcentrator technique followed by electrophoresis, and to compare the protein fractions obtained with those in serum. METHODS: CSF samples from 3 healthy dogs and 3 dogs with canine distemper virus infection were concentrated using a membrane microconcentrator having a 0.5 to 30,000 d nominal molecular weight limit (Ultrafree, Millipore, Billerica, MA, USA). Protein concentration was determined before and after concentration. Agarose gel electrophoresis was done on concentrated CSF samples, serum, and serial dilutions of one of the CSF samples. RESULTS: Electrophoretic bands were clearly identified in densitometer tracings in CSF samples with protein concentrations as low as 1.3 g/dL. The higher CSF protein concentration in dogs with distemper was mainly the result of increased albumin concentration. CONCLUSION: The microconcentrating method used in this study enables characterization of the main protein fractions in CSF by routine electrophoresis and may be useful for interpreting the underlying cause of changes in CSF protein concentrations.     

Paleo-Eskimo mtDNA genome reveals matrilineal discontinuity in Greenland

The Paleo-Eskimo Saqqaq and Independence I cultures, documented from archaeological remains in Northern Canada and Greenland, represent the earliest human expansion into the New World's northern extremes. However, their origin and genetic relationship to later cultures are unknown. We sequenced a mitochondrial genome from a Paleo-Eskimo human by using 3400-to 4500-year-old frozen hair excavated from an early Greenlandic Saqqaq settlement. The sample is distinct from modern Native Americans and Neo-Eskimos, falling within haplogroup D2a1, a group previously observed among modern Aleuts and Siberian Sireniki Yuit. This result suggests that the earliest migrants into the New World's northern extremes derived from populations in the Bering Sea area and were not directly related to Native Americans or the later Neo-Eskimos that replaced them.     

Geospatial variation of physical attributes and sugarcane productivity in cohesive soils

Precision Agriculture - Sugarcane productivity is highly variable across space, and knowledge about the spatial distribution of soil physical attributes is important to guide management practices...