Quantitative biomarkers for liver metastases: comparison of MRI diffusion-weighted imaging heterogeneity index and fluorine-18-fluoro-deoxyglucose standardised uptake value in hybrid PET/MR.

AIM: To investigate the ability of apparent diffusion coefficient (ADC) heterogeneity index to discriminate liver metastases (LM) from normal-appearing liver (NAL) tissue as compared to common magnetic resonance imaging (MRI) metrics, and to investigate its correlation with 2-[18F]-fluoro-2-deoxy-d-glucose (18F-FDG) positron-emission tomography (PET) standardised uptake value (SUV). MATERIALS AND METHODS: Thirty-nine liver metastases in 24 oncology patients (13 women, 11 men; mean age 56±13 years) with proven LM from heterogeneous sources were evaluated on a PET/MRI system. Abdominal sequences included Dixon and diffusion-weighted imaging (DWI) protocols with simultaneous PET. Tissue heterogeneity was calculated using the coefficient of variance (CV) of the ADC, and compared in LM and in NAL tissue of the same volume in an adjacent portion of the liver. The correlations between various ADC measures and PET SUV in distinguishing LM from NAL were evaluated. RESULTS: A good correlation was found between ADCcv and SUVpeak (r=0.712). Moderate inverse correlation was found between ADCmin and SUVpeak (r=-0.536), and a weak inverse correlation between ADCmean and SUVpeak (r=-0.273). There was a significant difference between LM and NAL when ADCcv (p<0.0001) and ADCmin (p=0.001) were used. Receiver operating characteristic (ROC) analysis of SUV, ADCcv, ADCmin, and ADCmean produced an AUC of 0.989, 0.900, 0.742, and 0.623 respectively. CONCLUSIONS: The ADCcv index is a potential biomarker of LM with better correlation to 18F-FDG PET SUVpeak than conventional MRI metrics, and may serve to quantitatively discriminate between LM and NAL.

Clinically applicable irreversible electroporation for eradication of micro-organisms.

Irreversible electroporation (IRE) damages cell membranes and is used in medicine for nonthermal ablation of malignant tumours. Our aim was to evaluate the antimicrobial effect of IRE. The pathogenic micro-organisms, Staphylococcus aureus, Streptococcus pyogenes, Escherichia coli, Pseudomonas aeruginosa and Candida albicans were subjected to IRE. Survival was measured as a function of voltage and the number of pulses applied. Combined use of IRE and oxacillin for eradication of Staph. aureus was also tested. Log10 reduction in micro-organisms positively correlated with the number of applied pulses. The colony count of Strep. pyogenes and E. coli declined by 3·38 and 3·05 orders of magnitude, respectively, using an electric field of 2000 V and 100 pulses. Killing of Staph. aureus and P. aeruginosa was achieved with a double cycle of IRE (2000, 1500 V and repeated 1250 V respectively) of 50-100 IRE pulses. The addition of subclinical inhibitory concentrations of oxacillin to the Staph. aureus suspension prior to IRE led to total bacterial death, demonstrating synergism between oxacillin and IRE. Our results demonstrate that using IRE with clinically established parameters has a marked in vitro effect on pathogenic micro-organisms and highlights the potential of IRE as a treatment modality for deep-seated infections, particularly when combined with low doses of antibiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: Irreversible electroporation (IRE) is utilized in interventional radiology to treat cancer patients. In this study we evaluated in vitro the antimicrobial effect of IRE. We demonstrated that using IRE with clinically established parameters has a marked effect on pathogenic micro-organisms and is synergistic to antimicrobials when both are combined. Our results point to the potential of IRE as a treatment modality for deep-seated infections.

Early 68GA-PSMA PET/MRI acquisition: assessment of lesion detectability and PET metrics in patients with prostate cancer undergoing same-day late PET/CT.

AIM: To compare lesion detectability and positron-emission tomography (PET) metric measurements between early-PET/magnetic resonance imaging (MRI) acquisition and same-day PET/computed tomography (CT). MATERIALS AND METHODS: The study was approved by the institutional review board and written informed consent was obtained from all patients. Twenty-one patients underwent non-time-of-flight (TOF) PET/MRI immediately following 68GA-prostate-specific membrane antigen (PSMA) tracer injection in two steps: firstly, early prostate PET/MRI (pPET/MRI) and early whole-body (WB) PET/MRI (wbPET/MR) followed by WB TOF PET/CT (wbPET/CT). Lesion detectability was compared between wbPET/MRI and wbPET/CT while PET metric measurements were compared between pPET/MR, wbPET/MRI, and wbPET/CT. RESULTS: Sixty-one and 63 lesions were found on wbPET/MRI and wbPET/CT, respectively (K=0.95, 95% confidence interval (CI)=0.89-1.0) with very good correlation between PET metric measurements (r=0.91; p=0.001). Bland-Altman plots demonstrated a mean percentage difference between wbPET/CT with wbPET/MRI of 34.4% with 95% limits of agreement (LOA) between -39% to 107.9% for metabolic tumour volume (MTV) and a mean difference of 30% with LOA between -13.4% to 73.4% for peak standardised uptake value (SUVpeak). CONCLUSION: Early PET/MRI demonstrates very good lesion detectability agreement and correlation with PET metrics compared to same-day PET/CT. Nevertheless, LOA are far beyond the clinically acceptable range, and therefore, PET/CT and early PET/MRI metrics cannot be used interchangeably.

New Mexico and the southwestern US are affected by a unique population of tomato spotted wilt virus (TSWV) strains.

Tomato spotted wilt virus (TSWV) is an important pathogen of many ornamental, greenhouse and agronomic crops worldwide. TSWV also causes sporadic problems in a number of crops in New Mexico (NM). Nucleocapsid gene sequences obtained from six different crop species across the state over four different years were used to characterize the NM TSWV population. This analysis shows that NM is affected by a unique TSWV population that is part of larger independent population present in the southwestern US. This population likely arose due to geographic isolation and is related to other TSWV populations from the US, Spain, and Italy.

A Coordinated Effort to Manage Soybean Rust in North America: A Success Story in Soybean Disease Monitoring.

Existing crop monitoring programs determine the incidence and distribution of plant diseases and pathogens and assess the damage caused within a crop production region. These programs have traditionally used observed or predicted disease and pathogen data and environmental information to prescribe management practices that minimize crop loss. Monitoring programs are especially important for crops with broad geographic distribution or for diseases that can cause rapid and great economic losses. Successful monitoring programs have been developed for several plant diseases, including downy mildew of cucurbits, Fusarium head blight of wheat, potato late blight, and rusts of cereal crops. A recent example of a successful disease-monitoring program for an economically important crop is the soybean rust (SBR) monitoring effort within North America. SBR, caused by the fungus Phakopsora pachyrhizi, was first identified in the continental United States in November 2004. SBR causes moderate to severe yield losses globally. The fungus produces foliar lesions on soybean (Glycine max) and other legume hosts. P. pachyrhizi diverts nutrients from the host to its own growth and reproduction. The lesions also reduce photosynthetic area. Uredinia rupture the host epidermis and diminish stomatal regulation of transpiration to cause tissue desiccation and premature defoliation. Severe soybean yield losses can occur if plants defoliate during the mid-reproductive growth stages. The rapid response to the threat of SBR in North America resulted in an unprecedented amount of information dissemination and the development of a real-time, publicly available monitoring and prediction system known as the Soybean Rust-Pest Information Platform for Extension and Education (SBR-PIPE). The objectives of this article are (i) to highlight the successful response effort to SBR in North America, and (ii) to introduce researchers to the quantity and type of data generated by SBR-PIPE. Data from this system may now be used to answer questions about the biology, ecology, and epidemiology of an important pathogen and disease of soybean.

First Report of Anthracnose of Sunflower Sprouts Caused by Colletotrichum acutatum in New Mexico.

In December 2011, edible sunflower sprouts (Helianthus annus) of two different commercially grown cultivars (Sungrown and Tiensvold) exhibiting stem and cotyledon lesions were submitted to the New Mexico State University Plant Clinic for disease diagnosis. The sample originated from an organic farm in Santa Fe County where the grower utilizes a small indoor growing facility. Stem lesions were elongate, reddish brown, and often constricted, resulting in stem girdling. Lesions on the cotyledons were dark brown with tan centers and round to irregular in shape. In some cases, the entire cotyledon was blighted. Fungal hyphae were observed on some lesions using a dissecting microscope. Colletotrichum acutatum was isolated from stem and cotyledon lesions when symptomatic tissue was plated on water agar. Conidia were fusiform ranging from 6.4 to 18.4 µm long and 2.1 to 5.1 µm wide and averaged 11.9 µm × 3.4 µm. Spores were measured from cream-colored colonies produced on acidified potato dextrose agar. PCR amplification and sequence analysis of 5.8S ribosomal DNA and internal transcribed spacers I and II was performed using primers ITS4 and ITS6 (2). An amplification product of approximately 600 base pairs in size was directly sequenced (GenBank Accession No. JX444690). A BLAST search of the NCBI total nucleotide collection revealed a 99% identity to multiple C. acutatum (syn: C. simmondsii) isolates. Four isolates were identified as C. acutatum based on morphological characteristics and DNA analysis. Koch's postulates were performed using four isolates of the pathogen and the two commercial sunflower cultivars (Sungrown and Tiensvold) originally submitted for disease analysis. Sunflower seeds were imbibed in distilled water for 24 h then sewn into peat plugs. Prior to seed germination, 5 ml of a C. acutatum spore solution (1 × 106/ml) from each isolate was applied to five peat plugs using an atomizer. Control plants were inoculated with distilled water and otherwise treated identically. Both sunflower cultivars were inoculated with each isolate of the pathogen and the test was replicated twice. The sewn peat plugs were incubated for 5 days at 21°C and 50% relative humidity. Symptoms similar to the original samples were present on 100% of the sprouts after 5 days. PCR and sequence analysis performed on cultures obtained from lesions showed a 100% match to the original New Mexico isolates fulfilling Koch's postulates. In an indoor organic facility, such as the one in NM, this disease has the potential to be very difficult to manage and the potential to infect a high percentage of the crop resulting in significant economic losses. To our knowledge, this is the second report of C. acutatum on sunflower sprouts in the United States (1) and the first report in New Mexico. References: (1) S. T. Koike et al. Plant Dis. 93:1351, 2009. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

First Report of Phytophthora nicotianae on Bulb Onion in the United States.

Phytophthora nicotianae (synonym P. parasitica) Breda de Haan was isolated from recently harvested onion bulbs (Allium cepa) in cold storage from a commercial field in southern New Mexico. Deteriorating, water-soaked tissue from the center of four bulbs was plated onto water agar and incubated at room temperature. After 72 h, cultures of Phytophthora (identified by the presence of coenocytic hyphae and papillate sporangia) were isolated and transferred to V8 agar amended with ampicillin (250 mg/liter), rifampicin (10 mg/liter), and pimaricin (0.2% wt/vol). Isolates were identified as P. nicotianae based on morphological characteristics and DNA analysis. Sporangia were sharply papilliate, noncaducous, and ovoid to spherical. The average sporangium size was 45.9 × 39.9 µm with a length-to-width ratio of 1.15. Clamydospores, both terminal and intercalary, were spherical to ovoid and averaged 37.2 × 35.2 µm (2). PCR from whole-cell extracts was performed on four cultured isolates from the infected onion tissue using previously described primers ITS4 and ITS6, which amplify the 5.8S rDNA and ITS1 and ITS2 internal transcribed spacers (1,4). A band of approximately 890 bp was amplified and directly sequenced (GenBank Accession No. HQ398876). A BLAST search of the NCBI total nucleotide collection revealed a 100% similarity to multiple P. nicotianae isolates previously sequenced (1). To confirm the pathogenicity of the isolates, onion seedlings were inoculated with 25 ml of P. nicotionae zoospore solution (15,000 zoospores/ml). Necrosis of leaf tissue and seedling death was observed 5 days postinoculation. P. nicotianae was reisolated from the infected onion seedlings and the ITS region was sequenced to confirm its identity. P. nicotianae was previously reported in bulb onion from Australia, Taiwan (Formosa), and Zimbabwe (Rhodesia) (2). P. nicotianae was reported on bunching onions (A. fistulosum) in Hawaii in 1989 (3). Onions are an important crop in New Mexico with a total production value of 47 million dollars in 2008 (NM Agriculture Statistics 2008). This discovery of a potentially significant postharvest disease poses a threat to the onion industry in New Mexico. To our knowledge, this is the first report of P. nicotianae in bulb onion in the United States and the first report of P. nicotianae in New Mexico on any crop. References: (1) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (2) D. C. Erwin and O. K. Ribeiro. Page 56 in: Phytophthora Diseases Worldwide. The American Phytopathological Society, St Paul, MN, 1996. (3) R. D. Raabe et al. Information Text Series No. 22. University of Hawaii. Hawaii Inst. Trop. Agric. Human Resources, 1981. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

First Report of Buckeye Rot Caused by Phytophthora nicotianae in Tomato in New Mexico.

Phytophthora nicotianae Breda de Haan was isolated from turning tomato fruit (Solanum lycopersicum L.) in August 2010 from a garden in central New Mexico. Symptoms typical of buckeye rot including brown, water-soaked, necrotic lesions with concentric rings were observed on three tomato fruit. Tissue from each fruit was surface sterilized and plated onto water agar and incubated at room temperature. After 72 h, colonies of Phytophthora (identified by the presence of coenocytic hyphae and papillate sporangia) were found and subcultured by hyphal tips to V8 agar amended with ampicillin (250 mg/liter), rifampicin (10 mg/liter), and pimaricin (0.2% wt/vol). The isolates of Phytophthora were identified as P. nicotianae based on morphological characteristics and DNA analysis. Sporangia were sharply papillate, noncaducous, and ovoid to spherical. The average sporangium size was 44.5 × 35.5 µm with a length-to-width ratio of 1.26. Chlamydospores, both terminal and intercalary, were spherical to ovoid and averaged 38.9 × 37.5 µm. PCR amplification and sequence analysis on three isolates from the infected tomato tissue was performed using primers ITS4 and ITS6 that amplify the 5.8S rDNA and ITSI and ITSII internal transcribed spacers (1,2). A band of approximately 890 bp was amplified and directly sequenced (GenBank Accession No. HQ711620). A BLAST search of the NCBI total nucleotide collection revealed a 100% similarity to multiple P. nicotianae isolates previously sequenced. Pathogenicity tests with sequenced P. nicotianae isolates were performed to confirm virulence on tomato fruit. Tomatoes were surface sterilized with 95% ethanol and 0.1 ml of a P. nicotianae zoospore suspension (10,000 zoospores/ml) or sterile water was pipetted onto the surface of the tomato fruit. After 5 days in a humidity chamber, all three inoculated tomatoes had expanding water-soaked, circular lesions and the negative control showed no disease symptoms. P. nicotianae was successfully reisolated from the inoculated tomato tissue and the ITS region was sequenced to confirm its identity. Although the disease has been reported in many other states since the early 1900s, to our knowledge, this is the first report of P. nicotianae causing disease on tomato in New Mexico. References: (1) D. E. L. Cooke and J. M. Duncan. Mycol. Res. 101:667, 1997. (2) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

First Report of Xylella fastidiosa in Peach in New Mexico.

Xylella fastidiosa is a gram-negative bacterium that causes disease in a wide variety of plants such as grapes, citrus trees, oleanders, and elm and coffee trees. This bacterium is xylem limited and causes disease symptoms such as leaf scorch, stunting of plant growth, branch dieback, and fruit loss. The presence of X. fastidiosa was previously reported in New Mexico where it was found to be infecting chitalpa plants and grapevines (3). In the summer of 2010, peach (Prunus persica (L.) Batsch) trees from two locations in northern New Mexico exhibited leaf deformity and stunting, dark green venation, slight mottling, and branch dieback. Preliminary viral diagnostic screening was performed by Agdia (Elkhart, IN) on one symptomatic tree and it was negative for all viruses tested. Three trees from two different orchards tested positive for X. fastidiosa by ELISA and PCR analysis using X. fastidiosa-specific primer sets HL (1) and RST (2). Bacterial colonies were also cultured from these samples onto periwinkle wilt media. Eight colonies obtained from these three plants tested PCR positive using the X. fastidiosa-specific primers. The 16S ribosomal and 16S-23S rRNA internal transcribed spacer (ITS) region (557 nucleotides) (GenBank Accession No. HQ292776) along with the gyrase region (400 nucleotides) (GenBank Accession No. HQ292777) was amplified from the peach total DNA samples and the bacterial colonies. Sequencing analysis of these regions indicate that the X. fastidiosa found in peach is 100% similar to other X. fastidiosa multiplex isolates including isolates from peach, pecan, sycamore, and plum trees and 99% similar to the X. fastidiosa isolates previously found in New Mexico. Further analysis of the 16S ribosomal and 16S-23S rRNA ITS sequences with maximum likelihood phylogenetic analysis using Paup also groups the peach isolates into the X. fastidiosa multiplex subspecies. The gyrase sequence could not be used to differentiate the peach isolates into a subspecies grouping because of the lack of variability within the sequence. This X. fastidiosa multiplex subspecies could possibly be a threat to the New Mexico pecan industry since pecan infecting X. fastidiosa isolates belong to the same bacterial subspecies. It is not known if X. fastidiosa subspecies multiplex isolates from peach are capable of infecting pecans but they are closely genetically related. It is interesting to note that the isolates from peach are different than previously described X. fastidiosa isolates in New Mexico that were infecting chitalpa and grapes (3). X. fastidiosa has previously been described in peach; the disease is called "phony peach". The peach trees exhibited stunting and shortened internodes as reported for "phony peach". They also exhibited slight mottling and branch dieback that may be due to the environment in New Mexico or perhaps they are also exhibiting mineral deficiency symptoms in association with the X. fastidiosa disease. To our knowledge, this is the first report of X. fastidiosa in peach in New Mexico. References: (1) M. H. Francis et al. Eur. J. Plant Pathol. 115:203, 2006. (2) G. V. Minsavage et al. Phytopathology 84:456, 1994. (3) J. J. Randall et al. Appl. Environ. Microbiol. 75:5631, 2009.

First Report of Septoria Leaf Spot of Pistachio in New Mexico.

In September of 2008, a Septoria sp., the causal agent of Septoria leaf spot of pistachio (Pistacia vera L.) was isolated from leaf lesions in an orchard in southern New Mexico. Tree fruit and nut crops including pistachios are becoming an increasingly important part of New Mexico's agricultural industry with total cash receipts of $103 million in 2007 (3). This preliminary positive for Septoria prompted a survey of pistachio-growing counties in the state. The surveyed orchards accounted for approximately 30% of the pistachio acreage in New Mexico. Results indicated that all five pistachio-growing counties had orchards infected with a Septoria sp. Isolates of Septoria from leaf lesions were identified as Septoria pistaciarum Caracc. based on the following symptoms and morphological characteristics of the fungus: leaf lesions were usually circular, 0.5 to 3 mm in diameter, and contained many pycnidia per lesion; pycnidia were dark, ostiolate, and measured 101 to 255 × 69 to 133 µm; and conidia were hyaline, filiform, contained 3 to 9 septa, and measured 3 to 4 × 60 to 149 µm. Most orchards were only mildly affected. In severe cases, hundreds of leaf lesions were present on diseased leaves; large sections of the leaves turned tan and some trees defoliated prematurely. This widespread occurrence of Septoria leaf spot in New Mexico in 2008 suggests that the disease had already been present in the state for several years. A higher average rainfall in the summer of 2008 provided excellent conditions for disease development. Because of the high amounts of inoculum currently present in New Mexico orchards, Septoria leaf spot may emerge as a recurring disease problem for pistachio producers. This disease was first reported in the United States in Texas in 1971 and was also reported in Arizona in 1989 (1,2,4). To our knowledge, this is the first report of Septoria leaf spot of pistachio in New Mexico. References: (1) A. Chitzandis. Ann. Inst. Phytopathol. Benaki 10:29, 1956. (2) J. L. Maas et al. Plant Dis. Rep. 55:72, 1971. (3) New Mexico Agricultural Statistics, Department of Agriculture, 2007. (4) D. J. Young and T. Michailides. Plant Dis. 73:775, 1989.

Dynamic Changes in Circulating MicroRNA Levels in Liver Cancer Patients Undergoing Thermal Ablation and Transarterial Chemoembolization.

BACKGROUND: Hepatic cancer patients who cannot undergo surgical resection of tumour are candidates for methods of interventional radiology - transarterial chemoembolization (TACE) or thermal ablative (TA) therapy. Both methods are causing characteristic changes in liver tissue (inflammatory immune response, hypoxia, elevated temperature, tissue destruction) which are accompanied with systemic secretion of cytokines or microRNAs (miRNAs). The aim of our study was to investigate whether the level of circulating miRNAs related to hypoxia (miR-21 and miR-210), liver injury (miR-122) and epithelial-mesenchymal transition (miR-200a) could reflect systemic effect of these intervention techniques. MATERIALS AND METHODS: Study consisted of 10 primary hepatocellular carcinoma patients treated with TACE and 10 patients with liver metastases of colorectal cancer treated with TA. Thermal ablation was performed using the radiofrequency or microwave generator (RITA, Microsulis, AngioDynamics,Inc), for TACE drug eluting beads (DCBeads, Biocompatibles Ltd.) were used. Tumours were evaluated using RECIST (Response Evaluation Criteria in Solid Tumours), mRECIST (modified RECIST) criterion and volumetry. For all patients we determined concentrations of miRNA in blood plasma samples from four time points (before intervention, immediately after intervention, 24 hours after intervention, 1 week after intervention) using TaqMan® Assays and quantitative real time polymerase chain reaction method. RESULTS: After both intervention techniques we observed changes in circulating miRNA levels. In TA cases we observed significant increase of miR-122 and miR-200a concentrations immediately after intervention, on the contrary in TACE we observed increase in miRNA concentration at time point 24 hours after intervention (miR-21, miR-210, miR-122, miR-200a). Increased concentration of circulating miRNA was followed by subsequent decline to initial level. These changes were consistent with presumed biological effect of TA and TACE on tumour tissue. CONCLUSION: Data of this pilot study show potential usage of circulating miRNA for monitoring of systemic effect of thermal ablative and intraarterial therapies. This work was created at Masaryk University as part of the project MUNI/A/1574/2018 and it was supported by Czech Ministry of Health grants No. 15-32484A, 16-31765A and 16-31314A. The authors declare they have no potential confl icts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 1. 3. 2019 Accepted: 4. 3. 2019.

Effect of suture material on tensile strength and complication rate in abdominal fascial defects repaired with acellular dermal matrix.

BACKGROUND: Repair of ventral hernias represents a challenging problem for surgeons. AlloDerm (LifeCell, Branchburg, NJ, USA), an acellular dermal matrix (ADM) product derived from cadaveric human skin, has gained in popularity in the management of abdominal hernias because of its ability to support neovascularization and therefore resist infection. Surgeons have traditionally used nonabsorbable suture when using ADM in this setting, perhaps because of concerns regarding wound strength. This study was undertaken to examine the influence of suture material on wound breaking strength and complication rates in abdominal wall defects closed with ADM. METHODS: Full-thickness abdominal defects were created in athymic rats and immediately repaired with an ADM interposition graft using either interrupted Prolene or Maxon suture. Complications were recorded over time and the animals were sacrificed at 1 month intervals. The abdominal repair complex was harvested and wound strength was measured using a tensiometer. RESULTS: There were no hernias in any of the groups. There were also no cases of major adhesions to the AlloDerm. Two rats in the Prolene group developed a stitch extrusion through the ventral skin. Wound breaking strength increased significantly from the second to third month after surgery in both suture groups (p=0.0000, LSD test). The breaking strength of the abdominal fascia-ADM complex increased over the course of the experiment in both test groups, but no significant difference was found between the groups at any time point (p=0.3157). At 3 months, the breaking strength of the Prolene group and Maxon group was nearly identical (27.1 N +/- SD 7.4 and 29.7 N +/- SD 9.6, respectively). CONCLUSIONS: We were unable to identify a significant difference in breaking strength at the interface between ADM and normal, native fascia when using permanent versus resorbable sutures.

Active specific immunotherapy in patients with melanoma. A clinical trial with mouse antiidiotypic monoclonal antibodies elicited with syngeneic anti-high-molecular-weight-melanoma-associated antigen monoclonal antibodies.

In two clinical trials the mouse antiidiotypic monoclonal antibody (MAb) MF11-30, which bears the internal image of human high-molecular-weight-melanoma-associated antigen (HMW-MAA) was administered by subcutaneous route without adjuvants to patients with stage IV malignant melanoma on day 0, 7, and 28. Additional injections were administered if anti-antiidiotypic antibodies were not found or their titer decreased. In the first phase I trial with 16 patients the initial dose was 0.5 mg per injection and escalated to 4 mg per injection. Neither toxicity nor allergic reactions were observed despite the development of anti-mouse Ig antibodies. Minor responses were observed in three patients. In a second clinical trial MAb MF11-30 was administered to 21 patients at a dose of 2 mg per injection, since this dose had been shown in the initial study to be effective in inducing anti-antiidiotypic antibodies. Two patients were inevaluable; in the remaining 19 patients, the average duration of treatment was 34 wk. In this trial as well, neither toxicity nor allergic reactions were observed. 17 of the 19 immunized patients increased the levels of anti-mouse Ig antibodies and 16 developed antibodies that inhibit the binding of antiidiotypic MAb MF11-30 to the immunizing anti-HMW-MAA MAb 225.28. One patient increased the level of anti-HMW-MAA antibodies. One patient achieved a complete remission with disappearance of multiple abdominal lymph nodes for a duration of 95 wk. Minor responses were observed in three patients. These results suggest that mouse antiidiotypic MAb that bear the internal image of HMW-MAA may be useful reagents to implement active specific immunotherapy in patients with melanoma.

Xylella fastidiosa Detected in New Mexico in Chitalpa, a Common Landscape Ornamental Plant.

Different strains of Xylella fastidiosa cause a variety of significant disease problems in agricultural and ornamental plants, including Pierce's disease in grapes, oleander leaf scorch, pecan bacterial leaf scorch, and alfalfa dwarf disease. X. fastidiosa has never been reported in New Mexico but is known to exist in surrounding states (California, Arizona, and Texas). During the summer of 2006, several chitalpa (Chitalpa tashkinensis) hybrid trees with leaf scorch symptoms and branch die back were observed in Las Cruces, NM and they tested positive for X. fastidiosa by ELISA. Additional samples from these plants and others were analyzed by ELISA, PCR (2), and cultured on XfD2 medium (1). Known positive and negative oleander samples from Arizona were included as controls. Fifteen of thirty tested chitalpa were PCR and ELISA positive, indicating that they were infected with X. fastidiosa. Bacterial colonies that were PCR positive were also recovered from 10 of the XF positive samples that were plated. DNA sequences of PCR products amplified from chitalpa and isolated bacterial colonies (GenBank Accession Nos. EF109936 and EF109937) were identical to each other, 97% similar to X. fastidiosa strain JB-USNA, and 96% similar to the Temecula 1 strain. Independent ELISA testing (Barry Hill, California Department Food and Agriculture, Sacramento, CA) confirmed our ELISA and PCR results. On the basis of these results, we conclude that X. fastidiosa is present in New Mexico and that the common landscape ornamental chitalpa is a host for X. fastidiosa. Additional work is required to determine if X. fastidiosa is pathogenic to chitalpa and to examine the relevance of this potential X. fastidiosa reservoir to agricultural production in New Mexico and other areas where chitalpa is grown. References: (1) R. P. P. Almeida et al. Curr. Microbiol. 48:368, 2004. (2) M. R. Pooler et al. Lett. Appl. Microbiol. 25:123, 1997.

Inositol trisphosphate stimulates calcium release from peeled skeletal muscle fibers.

The effects of inositol phosphates (tris (InsP3), bis (InsP2), mono (InsP)) on rabbit adductor magnus and soleus muscles were determined using mechanically peeled fibers (sarcolemma removed). Isometric force generation of each fiber was continuously monitored and was used along with 45Ca to detect calcium release from internal fiber stores. All experiments were conducted at a physiological Mg2+ concentration (10(-3) M) of the bathing solutions. The inositol phosphates did not directly activate the contractile apparatus. At bath concentrations of 100-300 microM, only InsP3 was capable of stimulating Ca2+ release. In contrast, 1 microM InsP3 maximally and selectively stimulated Ca2+ release when microinjected into the myofilament lattice. Calcium releasing effects of InsP2 and InsP were manifested at 10 microM when they were microinjected. The end-to-end internal Ca2+ release and subsequent fiber force generation stimulated by the locally applied microinjected InsP3 suggests that the InsP3-induced Ca2+ release mechanism may involve propagation, but not via the Ca2+-induced Ca2+ release, since procaine did not inhibit this response. These findings support the possibility that InsP3 plays a role in skeletal muscle excitation-contraction coupling.

Genetic and Phenotypic Variation of the Pepper golden mosaic virus Complex.

ABSTRACT Three isolates of the bipartite begomovirus Pepper golden mosaic virus (PepGMV) were characterized for genomic and biological properties. The complete nucleotide sequences of the DNA-A and DNA-B components were determined from infectious clones of PepGMV-Serrano (PepGMV-Ser), PepGMV-Mosaic (PepGMV-Mo), and PepGMV-Distortion (PepGMV-D). Nucleotide sequence identity among PepGMV components ranged from 91 to 96% for DNA-A and from 84 to 99% for DNA-B, with each PepGMV component most closely related to the corresponding component of Cabbage leaf curl virus (CaLCV). However, phylogenetic relationships among begomovirus components were incongruent because DNA-A of PepGMV and CaLCV share an inferred evolutionary history distinct from that of DNA-B. The cloned components of PepGMV-Ser, -Mo, and -D were infectious by biolistic inoculation to pepper but differed in symptom expression: PepGMV-Ser exhibited a bright golden mosaic, PepGMV-Mo produced a yellow-green mosaic, and PepGMV-D caused only a mild mosaic and foliar distortion followed by a "recovery" phenotype in which leaves developing after initial symptom expression appeared normal. Differences in symptoms also were observed on tomato, tobacco, and Datura stramonium. Progeny virus derived from clones of PepGMV-Ser and -Mo were transmitted from pepper to pepper by the B biotype of Bemisia tabaci; progeny virus derived from PepGMV-D clones was not transmissible by the B biotype. Reassortant genomes derived from heterologous DNA components of the three isolates were infectious in all possible pairwise combinations, with symptom phenotype in pepper determined by the DNA-B component. Collectively, these results indicate that the three virus isolates examined may be considered distinct strains of PepGMV that have the capacity to exchange genetic material.

Hyperprolactinemia in multiple endocrine adenomatosis, type 1.

Three cases are described in which hyperprolactinemia occurred as a feature of multiple endocrine adenomatosis, type 1 (MEA-1); enlargement of the sella turcica varied from gross to absent, and serum prolactin (PRL) levels ranged from 21 to 1,000 ng/ml in these cases. Since PRL-secreting pituitary tumors may occur with variable presentation in MEA-1, periodic measurements of serum PRL levels should be carried out to detect this abnormality.

Cyclic GMP metabolism in psoriasis: activation of soluble epidermal guanylate cyclase by arachidonic acid and 12-hydroxy-5,8,10,14-eicosatetraenoic acid.

Soluble guanylate cyclase activity was measured in normal and psoriatic human epidermis. The specific activity of guanylate cyclase was determined to be increased 10-fold and 3-fold in involved and uninvolved epidermis of psoriatics, respectively, compared to normal epidermis. Arachidonic acid (5 to 100 micrometers) or 12-hydroxy-5,8,10,14-eicosatetraenoic acid (HETE) (5 to 50 micrometers) stimulated guanylate cyclase activity from involved epidermis 2- to 3-fold and from uninvolved epidermis up to 2-fold, but these fatty acids had no effect on the activity of this cyclase from normal epidermis. These results indicate that there is an increase in the cGMP biosynthetic capacity of involved epidermis from psoriatics that derives from a markedly increased specific activity of guanylate cyclase and an alteration in a property of this enzyme activity which renders it responsive to fatty acids reported to accumulate in this lesion. These observations are consistent with the report that an elevated steady-state level of cGMP is one of the consequences of the strikingly altered metabolism of cGMP in psoriatic epidermis.

Mapping the Down syndrome chromosome region. Establishment of a YAC contig spanning 1.2 megabases.

The triplication of a region of chromosome 21 around D21S55 in 21q22.2-22.3 has been involved in the main features of Down syndrome including mental retardation (Down syndrome chromosome region: DCR). To improve the physical map of this region, we screened yeast artificial chromosome (YAC) libraries with ETS2 and ERG sequences. Five selected clones were analyzed by AluPCR, pulsed-field gel electrophoresis, and in situ hybridization. A 1.2-Mg contig, encompassing the protooncogenes ETS2 and ERG, was identified, its restriction map established and compared to the genomic map. ERG is distal to D21S55 and proximal to ETS2. ERG and ETS2 genes are 400 kb apart and in opposite orientations. The contig contains the distal boundary and part of the DCR. Three putative HTF islands were identified.

A technique for determining E rosette levels by cytofluorographic analysis.

A method is described for determining the percentage of rosettes formed by sheep erythrocytes and human peripheral lymphocytes (ER) using cytofluorographic analysis (CFGA). The technique utilizes acridine orange dye for differentially staining nucleus and cytoplasm of the lymphocytes to distinguish them from erythrocytes, and glutaraldehyde for fixation of the rosettes. This technique was compared with light microscope counting (LMC) or ER. CFGA gave similar results with better reproducibility than LMC, entailed less time for counting, and markedly reduced operator fatigue.