Segmentation of 3D images of plant tissues at multiple scales using the level set method

Background

Developmental biology has made great strides in recent years towards the quantification of cellular properties during development. This requires tissues to be imaged and segmented to generate computerised versions that can be easily analysed. In this context, one of the principal technical challenges remains the faithful detection of cellular contours, principally due to variations in image intensity throughout the tissue. Watershed segmentation methods are especially vulnerable to these variations, generating multiple errors due notably to the incorrect detection of the outer surface of the tissue.

Results

We use the level set method (LSM) to improve the accuracy of the watershed segmentation in different ways. First, we detect the outer surface of the tissue, reducing the impact of low and variable contrast at the surface during imaging. Second, we demonstrate a new edge function for a level set, based on second order derivatives of the image, to segment individual cells. Finally, we also show that the LSM can be used to segment nuclei within the tissue.

Conclusion

The watershed segmentation of the outer cell layer is demonstrably improved when coupled with the LSM-based surface detection step. The tool can also be used to improve watershed segmentation at cell-scale, as well as to segment nuclei within a tissue. The improved segmentation increases the quality of analysis, and the surface detected by our algorithm may be used to calculate local curvature or adapted for other uses, such as mathematical simulations.

     

Perivascular wall tumour presenting as pastern mass in a Standardbred gelding

A 2-year-old Standardbred gelding was referred for a mass on the palmaromedial right front pastern which was accompanied by progressively worsening lameness. The mass was firm to palpation and covered by normal skin. Ultrasonographically, a smooth encapsulated mass was present, medial to the flexor tendons and palmar to the neurovascular bundle. Because of a poor prognosis for future athletic performance without surgical or chemotherapeutic intervention and economic constraints preventing further diagnostics and treatment, the horse was euthanised. Post-mortem magnetic resonance imaging, histopathology and immunohistochemistry revealed the mass to be a perivascular wall tumour, the first record of such a neoplasia in the horse.     

Comparison of oil and phytosterol levels in germplasm accessions of corn, teosinte, and Job's tears

Seeds of 49 accessions of corn (Zea mays ssp. mays), 9 accessions of teosinte (Zea species that are thought to be ancestors and probable progenitors to corn), and 3 accessions of Job's tears (Coix lacryma), obtained from a germplasm repository, were ground and extracted with hexane. Whole kernel oil yields and levels of four phytonutrients (free phytosterols, fatty acyl phytosterol esters, ferulate phytosterol esters, and gamma-tocopherol) in the oils were measured. Among the seeds tested, oil yields ranged from 2.19 to 4.83 wt %, the levels of ferulate phytosterol esters in the oil ranged from 0.047 to 0.839 wt %, the levels of free phytosterols in the oil ranged from 0.54 to 1.28 wt %, the levels of phytosterol fatty acyl esters in the oil ranged from 0.76 to 3.09 wt %, the levels of total phytosterols in the oil ranged from 1.40 to 4.38 wt %, and the levels of gamma-tocopherol in the oil ranged from 0.023 to 0.127 wt %. In general, higher levels of all three phytosterol classes were observed in seed oils from accessions of Zea mays ssp. mays than in seed oils from accessions of the other taxonomic groups. The highest levels of gamma-tocopherol were observed in teosinte accessions.     

SENSITIVITY AND SPECIFICITY OF RADIOGRAPHIC DETECTION OF CANINE ELBOW INCONGRUENCE IN AN IN VITRO MODEL

Canine elbow incongruence is believed to be the consequence of underdevelopment of the radius. The purpose of this study is to determine the sensitivity and specificity of radiography to detect elbow incongruence in an in vitro model and to assess the optimal elbow angle and radiographic beam position. Five normal cadaveric canine left forelimbs were used. A four-pin, type 1 external fixator with a linear motor side bar was fixed to the cranial part of the radius of each limb and a 1 cm segment of bone was removed from the mid-diaphysis to allow radial shortening. Each elbow was subjected to the same protocol. They were radiographed at two different angles (90 degrees and 135 degrees) of flexion, with 10 different radiographic beam positions (centered on the humeral condyle, 3 cm cranial, 3 cm caudal, 3 cm distal, 3 cm proximal, 3 cm cranio-proximal, 3 cm cranio-distal, 3 cm caudo-proximal, 3 cm caudo-distal and on the shoulder joint) and at four different level of radial shortening (0, 1, 2 and 3 mm). In addition, a radiographic view centered on the elbows flexed at 135 degrees was made after simulating weight bearing. The acquired digital images were independently evaluated by three evaluators unaware of the elbows status. The elbows were judged normal, incongruent or borderline based on specific criteria. The sensitivity for detection of elbow incongruence at and beyond 2 mm was excellent at 90 degrees (median = 100% for all views) and good at 135 degrees (median = 80%) of flexion with no difference between examiners. The sensitivity at 1 mm of incongruence was unchanged at 135 degrees but was reduced at 90 degrees of flexion (median = 60%) with a significant difference between the evaluators. The specificity was significantly different between the evaluators and ranged from 70% to 90% at 90 degrees of flexion and from 50% to 80% at 135 degrees. The lowest specificities at 90 degrees were obtained with the proximal displacements of the X-ray beam. Simulating weight bearing significantly decreased the sensitivity at 1 mm (from 80% to 50%) and 3 mm (from 100% to 80%) of incongruence and slightly increased the specificity (from 55% to 65%). Radiography is a sensitive and specific test to detect moderate-to-severe radio-ulnar incongruence (2 mm and over) if the elbow is flexed at a 90 degrees angle regardless of the radiographic beam position. Finally, canine elbow incongruence appeared reduced after an in vitro weight-bearing simulation.     

Insecticide resistance in cotton aphid Aphis gossypii (Glov.) in the Sudan Gezira

The resistance to insecticides of three Sudanese strains of A. gossypii (Glov.) collected from cotton fields in the Sudan Gezira Scheme over three seasons (1988, 1989, 1990) and that of two French strains was studied in the laboratory. When compared with a known susceptible strain, the aphids were found to be resistant-to the eight insecticides tested. Evolution of resistance in Sudanese strains during the three crop seasons was observed. Assay of aphid homogcnate for carboxytesterase activity towards the substrates α-naphthyl acetate and β-naphthyl acetate showed that there was no enhancement of this class of enzyme and thus it was not a cause of resistance in this species. A study of interaction between the acetylcholinesterase (AchE) and pirimicarb established the kinetics of the inhibition process. I₅₀ values were found to be much higher for the Sudanese strains than for the susceptible strain. First-order inhibition kinetics revealed that resistance towards pirimicarb in Sudanese-strains was caused by a modified AchE which had a reduced affinity (higher K_a value) and poor carbamylation ability (lower K₂ value) for pirimicarb. The resistance mechanisms for the other insecticides remain to be studied.     

Massive transfusion in dogs: 15 cases (1997-2001)

OBJECTIVE: To determine clinical characteristics of dogs that received massive transfusion and identify the underlying diseases, complications, and outcomes. DESIGN: Retrospective study. ANIMALS: 15 dogs. PROCEDURE: Medical records of dogs receiving a massive blood transfusion were evaluated for transfusion volume, underlying disease process or injury, benefits and complications of transfusion, and outcome. A massive transfusion was defined as transfusion of a volume of blood products in excess of the patient's estimated blood volume (90 ml/kg [40 ml/lb]) in a 24-hour period or transfusion of a volume of blood products in excess of half the patient's estimated blood volume in a 3-hour period. RESULTS: Six dogs had intra-abdominal neoplasia resulting in hemoabdomen, 3 had suffered a traumatic incident resulting in hemoabdomen, and 6 had non-traumatic, non-neoplastic blood loss. Mean volumes of packed RBC and fresh-frozen plasma administered were 66.5 ml/kg (30 ml/lb) and 22.2 ml/kg (10 ml/lb), respectively. All dogs evaluated developed low ionized calcium concentrations and thrombocytopenia. Transfusion reactions were recognized in 6 dogs. Four dogs survived to hospital discharge. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that massive transfusion is possible and potentially successful in dogs. Predictable changes in electrolyte concentrations and platelet count develop.