Evidence for vertical transmission of Mycoplasma haemocanis, but not Ehrlichia ewingii, in a dog.

A 2-year-old female intact pregnant Beagle was evaluated after the owner surrendered her to a shelter. Prepartum and 2 months postpartum at the time of routine spay, the dam was whole-blood polymerase chain reaction (PCR) positive for Ehrlichia ewingii. She was also whole-blood PCR positive for Mycoplasma haemocanis prepartum and continuously for 5 months thereafter. The dam delivered 5 healthy puppies, 1 of which was whole-blood PCR positive for M. haemocanis. All 5 puppies had antibodies against Ehrlichia spp. at 1 month of age but not thereafter, and all puppies were Ehrlichia spp. PCR negative for 5 months of follow-up. Therefore, this study supports a potential role for vertical transmission in the maintenance of M. haemocanis in dogs as reservoir hosts. In contrast, in this case there was no evidence that E. ewingii was transmitted transplacentally or during the perinatal period.

A Case of a Paediatric Patient With Allergic Contact Dermatitis to Benzoyl Peroxide.

Allergic contact dermatitis to benzoyl peroxide can occur in up to 6.5% of those with a history of exposure to this potential allergen. Conversely, irritant contact dermatitis is very common with benzoyl peroxide and can be differentiated from allergic contact dermatitis based on the patient's history and clinical signs and symptoms. We present a case of a paediatric patient with patch test-confirmed severe allergic contact dermatitis to benzoyl peroxide requiring hospitalisation and systemic treatment.

Harmonic subtraction for evaluating right ventricle ejection fraction from planar equilibrium radionuclide angiography.

We report an initial investigation of a subtraction-based method to estimate right ventricle ejection fraction (RVEF) from ECG-gated planar equilibrium radionuclide angiography (ERNA) data. Twenty-six consecutive patients referred for scintigraphic evaluation of cardiac function prior to chemotherapy had ECG-gated first-pass (FP) imaging and ERNA imaging performed following the same radiotracer injection. RVEF was computed from FP images (RVEFFP) and separately from ERNA images (RVEFERNA). Standard methods for computing ejection fractions were used to obtain RVEFFP values. RVEFERNA values were obtained using harmonic subtraction of the left ventricular contribution from a biventricular region of interest contoured on the equilibrium images acquired in the shallow right anterior oblique projection. Clinically acquired chest CT data were used to derive information regarding the relative position of the left and right ventricle and about the presence of pulmonary artery enlargement. Computation of RVEFERNA was successful for each of the 26 patients. Computation of RVEFFP failed for four patients. For the 22 patients for which RVEF was computed using both methods, the average RVEFFP was 49% and the average RVEFERNA was 51%, with coefficients of variation of 11 and 7.5%, respectively. Low RVEFERNA values were associated with pulmonary artery dilation. Estimation of RVEFERNA, using a harmonic subtraction-based method of computation is clinically feasible and accurate in the patient population studied. The results support further investigation in patients with frank heart failure.

Clinical intervention for quality improvement of gastric-emptying studies.

UNLABELLED: Prompted by clinical concerns for false-negative tests, we implemented a clinical intervention consisting of a training session and an image-based verification procedure to document homogeneous radioactivity distribution in the radiolabeled meal (egg substitute per the guideline). METHODS: A technologist training session emphasized the importance of thorough mixing of (99m)Tc-sulfur colloid in the egg meal. For 6 mo after training, an image of the prepared mixed egg was acquired before patient ingestion. Consecutive gastric-emptying studies performed 6 mo before and after training were reviewed by 2 experienced physicians. RESULTS: There were 7 abnormal and 44 normal studies before and 15 abnormal and 29 normal studies after training (P < 0.05). Subjective evaluations of images for meal-mixing quality by 2 readers correlated with each other and with an objective measure of expected gastric-emptying physiology (correlation coefficients, 0.54 and 0.38, respectively). CONCLUSION: The described clinical intervention improved the accuracy of our gastric-emptying studies by decreasing false-negative studies.

The EANM and SNMMI practice guideline for lymphoscintigraphy and sentinel node localization in breast cancer.

PURPOSE: The accurate harvesting of a sentinel node in breast cancer includes a sequence of procedures with components from different medical specialities, including nuclear medicine, radiology, surgical oncology and pathology. The aim of this document is to provide general information about sentinel lymph node detection in breast cancer patients. METHODS: The Society of Nuclear Medicine and Molecular Imaging (SNMMI) and the European Association of Nuclear Medicine (EANM) have written and approved these guidelines to promote the use of nuclear medicine procedures with high quality. The final result has been discussed by distinguished experts from the EANM Oncology Committee, the SNMMI and the European Society of Surgical Oncology (ESSO). CONCLUSION: The present guidelines for nuclear medicine practitioners offer assistance in optimizing the diagnostic information from the SLN procedure. These guidelines describe protocols currently used routinely, but do not include all existing procedures. They should therefore not be taken as exclusive of other nuclear medicine modalities that can be used to obtain comparable results. It is important to remember that the resources and facilities available for patient care may vary.

Status of sentinel lymph node for breast cancer.

Long-awaited results from randomized clinical trials designed to test the validity of sentinel lymph node biopsy (SLNB) as replacement of axillary lymph node dissection (ALND) in management of early breast cancer have recently been published. All the trials conclude SLNB has survival rates comparable to those of ALND (up to 10 years in one study) and conclude SLNB has less morbidity than ALND. All the trials support replacing ALND with SLNB for staging in early breast cancer; all support SLNB as the standard of care for such cancer. The SLNB protocols used in the trials varied, and no consensus that would suggest a standard protocol exists. The results of the trials and of other peer-reviewed research do, however, suggest a framework for including some specific methodologies in accepted practice. This article highlights the overall survival and disease-free survival data as reported from the clinical trials. This article also reviews the status of SLN procedures and the following: male breast cancer, the roles of various imaging modalities (single-photon emission computed tomography/computed tomography, positron emission tomography/computed tomography, and ultrasound), ductal carcinoma in situ, extra-axillary SLNs, SLNB after neoadjuvant chemotherapy, radiation exposure to patients and medical personnel, and a new radiotracer that is the first to label SLNs not by particle trapping but by specific macrophage receptor binding. The proper Current Procedural Terminology (CPT) code for lymphoscintigraphy and SLN localization prior to surgery is 78195.

Microsclerotia development in Verticillium dahliae: Regulation and differential expression of the hydrophobin gene VDH1.

The vascular wilt fungus Verticillium dahliae produces persistent resting structures, known as microsclerotia, which are important for this plant pathogen's long-term survival. Previously, we identified a hydrophobin gene (VDH1) that is necessary for microsclerotial production. The current study of VDH1's expression, and its regulation, was undertaken to provide insight into the largely uncharacterized molecular mechanisms relevant to microsclerotial development. Reporter gene analysis showed that VDH1 is specifically expressed in developing microsclerotia, as well as in hyphal fusions and conidiophores, suggesting that VDH1 mediates the development of microsclerotia from conidiophores and other hyphal structures. We report also on the effects of nutrient availability on the regulation of microsclerotial development in V. dahliae; the gene's activity appears to be regulated in response to carbon availability. Lastly, constitutive expression of VDH1 results in delayed disease symptom development, but has no noticeable effect on in vitro microsclerotial development.

Monitoring renal function: a prospective study comparing camera-based technetium-99m mercaptoacetyltriglycine clearance and creatinine clearance.

OBJECTIVES: To determine the value and limitations of technetium-99m mercaptoacetyltriglycine (MAG3) clearance measurements obtained using a gamma camera (camera-based MAG3 clearance), a prospective study was conducted to evaluate the reproducibility of camera-based MAG3 clearance compared with that of conventional creatinine clearance. METHODS: A total of 24 male patients with stable renal function were entered into the study. The mean age was 66.5 +/- 7.9 years, and the mean serum creatinine was 1.38 +/- 0.57 mg/dL. MAG3 renal scans and 24-hour creatinine clearance measurements were performed 11 +/- 8 days apart. A camera-based MAG3 clearance was obtained at each MAG3 scan; no blood samples were required. Bland-Altman plots were constructed to assist in data analysis. RESULTS: The Pearson correlation for the first and second camera-based MAG3 clearances (mean 151 versus 158 mL/min/1.73 m2, respectively) was 0.965 compared with 0.729 for the two creatinine clearance measurements (mean 62 versus 72 mL/min/1.73 m2, respectively). Even with the omission of two outliers, the creatinine clearance would have to change by 58.2% compared with the baseline measurement before the clinician could be confident the change exceeded the error of measurement. In contrast, the change required for the camera-based MAG3 clearance was 30.8%. CONCLUSIONS: The results of this study have demonstrated that MAG3 clearance obtained using a camera-based technique shows greater precision than the conventional creatinine clearance and is superior to the conventional creatinine clearance for monitoring changes in renal function.

Comparison of camera-based 99mTc-MAG3 and 24-hour creatinine clearances for evaluation of kidney function.

OBJECTIVE: The 24-hour creatinine clearance is the standard clinical technique for measuring kidney function; however, this measurement is cumbersome and inconvenient for patients. We hypothesized that a camera-based technetium-99m mercaptoacetyltriglycine (MAG3) clearance obtained simultaneously with a standard MAG3 scan would correlate well with the 24-hour creatinine clearance and could serve as a simple marker of kidney function. MATERIALS AND METHODS: Data were obtained from a retrospective analysis of 28 patients with varying degrees of kidney dysfunction and 85 subjects evaluated for kidney donation. The MAG3 clearance was calculated using a camera-based technique without blood or urine sampling. The creatinine clearance was measured using the plasma creatinine and a 24-hour urine collection. The MAG3 and creatinine clearances were corrected for body surface area, and clearance values in healthy subjects and patients were compared using the paired Student's t test. The linear association between the MAG3 and creatinine clearances was expressed by Pearson's correlation coefficient. RESULTS: The mean MAG3 clearance in the potential kidney donors was 321 +/- 95 mL/min/1.73 m2 (95% CI, 171-546 mL/min/1.73 m2), significantly higher than the mean creatinine clearance of 152 +/- 51 mL/min/1.73 m2 (79-278 mL/min/1.73 m2, p < 0.001). The mean MAG3 clearance in patients was 153 +/- 70 mL/min/1.73 m2 (32-316 mL/min/1.73 m2) and was also significantly higher than the mean creatinine clearance of 74 +/- 36 mL/min/1.73 m2 (21-138 mL/min/1.73 m2, p < 0.001). The ratio of the mean creatinine clearance to the mean MAG3 clearance was essentially the same for volunteers and patients, 0.47 and 0.48, respectively. The Pearson's correlation between the MAG3 and creatinine clearances was 0.80 (0.72-0.86). CONCLUSION: The camera-based 99mTc-MAG3 clearance correlates well with the 24-hour creatinine clearance and can provide a simple and convenient index of kidney function.

Intraoperative gamma imaging of axillary sentinel lymph nodes in breast cancer patients.

Sentinel lymph node (SLN) biopsy is now standard practice in the management of many breast cancer patients. Localization protocols vary in complexity and rates of success. The least complex involve only intraoperative gamma counting of radiotracer uptake or intraoperative visualization of blue-dye uptake; the most complex involve preoperative gamma imaging, intraoperative counting and intraoperative dye visualization. Intraoperative gamma imaging may improve some protocols. This study was conducted to obtain preliminary experience and information regarding intraoperative imaging. Sixteen patients were enrolled: 8 in a protocol that included intraoperative counting and dye visualization (probe/dye), 8 in a protocol that involved intraoperative imaging, counting and dye visualization (camera/probe/dye). Preoperative imaging of all 16 patients was performed using a GE 500 gamma camera with a LEAP collimator (300 cpm/muCi). The results of this imaging were not, however, given to the surgeon until the surgeon had completed the procedures required for the study. A Care Wise C-Trak probe was used for intraoperative counting. A Gamma Medica Inc. GammaCAM/OR (12.5 x 12.5 cm FOV) with a LEHR collimator (135 cpm/muCi) was used for intraoperative imaging. Times from start of surgery to external detection of a radioactive focus and to completion of excision of SLNs were recorded. Foci were detected preoperatively via imaging in 16/16 patients. Intraoperative external detection using the probe was accomplished in less than 4 min (mean = 1.5 min) in 15/16 patients, and via intraoperative imaging in 6/8 patients. The average time for completion of excision of nodes was 19 min for probe/dye and 28 min for camera/probe/dye. In one probe/dye case, review of the preoperative images prompted the surgeon to resume axillary dissection and remove one additional SLN.

Mutations in VMK1, a mitogen-activated protein kinase gene, affect microsclerotia formation and pathogenicity in Verticillium dahliae.

Verticillium dahliae is an important soil-borne fungal pathogen that causes vascular wilt diseases in a large variety of important crop plants. Due to its persistence in the soil, control of Verticillium wilt relies heavily on soil fumigation. The global ban on methyl bromide, a highly effective soil fumigant, poses an urgent need to develop alternative control measures against Verticillium wilt; and these might be more forthcoming with a better understanding of the molecular and cellular mechanisms that underpin the pathogenicity of V. dahliae. In this study, we assessed the role in growth, development, and pathogenicity of VMK1, a gene encoding a mitogen-activated protein (MAP) kinase (hence, Verticillium MAP Kinase 1). Disruption of VMK1 via Agrobacterium tumefaciens-mediated transformation, in two V. dahliae isolates, one from lettuce and the other from tomato, resulted in severely reduced virulence in diverse host plants, suggesting that VMK1 is essential for pathogenicity and that the MAP kinase-mediated signaling pathway has a conserved role in fungal pathogenicity. The vmk1 mutants also exhibited reduced conidiation and microsclerotia formation, suggesting that the gene is important for multiple cellular processes.

Cloning and targeted disruption, via Agrobacterium tumefaciens-mediated transformation, of a trypsin protease gene from the vascular wilt fungus Verticillium dahliae.

A gene encoding a trypsin protease was isolated from a tomato isolate of Verticillium dahliae. The gene, designated VTP1, contains two introns and is predicted to encode a protein of 256 amino acids. The gene is present in V. dahliae isolates from different host plants and in V. albo-atrum; weakly hybridizing sequences are present in V. tricorpus. VTP1 cDNA sequences were identified in a sequence tag analysis of genes expressed under growth conditions that promote microsclerotia development. Replacement of the gene, by Agrobacterium tumefaciens-mediated transformation (ATMT), with a mutant allele construct did not noticeably alter either pathogenicity or growth in culture. Searches of expressed sequence tag databases showed that, in addition to the VTP1 gene, V. dahliae contains two genes encoding subtilisin-like proteases similar to those produced by pathogenic Aspergillus spp. This is the first description of the application of ATMT to the molecular analysis of phytopathogenic Verticillium spp.

Immunohistochemical evaluation of sentinel lymph nodes in breast carcinoma patients.

Sentinel lymph node sampling has become an alternative to axillary lymph node dissection to provide prognostic and treatment information in breast cancer patients. The role of immunohistochemistry has yet to be established. A total of 241 sentinel lymph nodes (in 270 slides) from 91 patients with invasive carcinoma (73 ductal, 9 lobular, 8 mixed lobular/ductal, 1 NOS) were studied for presence of macrometastases (> 0.2 cm), identified in hematoxylin and eosin sections, and occult metastases (micrometastases [< or = 0.2 cm], clusters of cells, isolated carcinoma cells), identified only by immunohistochemistry. Intraoperative touch preparations, frozen sections, seven hematoxylin and eosin levels (L1-L7), and two AE1-3 cytokeratin immunohistochemistries (L1, L4-5) of the entire bisected or trisected sentinel lymph node were examined. Thirty-one (34%) patients had 50 positive sentinel lymph nodes. Twenty-six (33%) sentinel lymph nodes had metastatic carcinoma (11 macrometastases, 11 micrometastases, 3 clusters of cells, 1 isolated carcinoma cells) by touch preparations, frozen sections, and one hematoxylin and eosin (L1). Thirty-eight (43%) were positive by AE1-3 immunohistochemistry (L1) (11 macrometastases, 8 micrometastases, 13 clusters of cells, 6 isolated carcinoma cells), significantly more than by touch preparations, frozen sections, hematoxylin and eosin L1, or hematoxylin and eosin L2-7. Cytokeratin immunostain on L4-5 demonstrated 31 (34%) positive sentinel lymph nodes, a similar frequency to cytokeratin immunostain on L1. Size of sentinel lymph node metastasis did not correlate with size, histologic grade, or type of primary breast carcinoma. AE1-3 (L1) immunohistochemistry is highly sensitive in delineating sentinel lymph node metastasis, especially clusters of cells and isolated carcinoma cells. The prognostic significance of clusters of cells and isolated carcinoma cells and the value of AE1-3 immunohistochemistry on frozen sections need to be determined.