Frequency of Alternaria Genotypes Resistant to SDHI Fungicides in California Pistachio Orchards Determined by Real-Time PCR.

Real-time PCR methods were developed to quantify the frequency of SDHC-H134R and SDHB-H277Y mutants associated with succinate dehydrogenase inhibitors (SDHI) resistance in Alternaria populations from pistachio. The linearity of the standard curves demonstrated the applicability in the quantification of the assays. The accuracy and reliability of the qPCR protocols to determine the frequency of mutants in real samples were corroborated. Orchards visibly affected by Alternaria late blight were sampled. The frequency of mutants was determined using the qPCR assays, while the frequency of resistant phenotypes was determined using a single discriminatory dose. The statistical analysis showed that the frequencies of the mutation SDHC-H134R determined with the qPCR assay were highly correlated with those estimated with the conventional method. The survey also evidenced that resistance to boscalid is still widespread in California. Results also indicated the possible contribution of other mutations to SDHI resistance. Our results confirmed the prevalence of SDHC-H134R mutants and the occurrence of mutation SDHB-H277Y at low frequencies. The real-time PCR methods developed in this study were able to detect differences in the frequencies of resistant mutants caused by the use of chemical fungicides. Finally, the effects of two fungicide programs on the frequency of mutants resistant to SDHI and quinone outside inhibitors fungicides were studied using qPCR assays. The experiments demonstrated that the use of anilinopyrimidine and demethylation inhibitors fungicides in the same program reduced the frequency of these mutations in Alternaria populations. The qPCR methods developed and used in this study can be used to track resistance levels in the pistachio orchards on a large scale.

Characterization, pathogenicity, and fungicide sensitivity of Alternaria isolates associated with preharvest fruit drop in California citrus.

Alternaria rot has been recently described as an emerging fungal disease of citrus causing significant damage in California groves. A survey was conducted to determine latent infections on fruits, twigs, and leaves and investigate their seasonal patterns during 2019 and 2020. On fruits, latent infections were more associated with the stem end than with the stylar end, except during spring when a significantly high percentage of flowers (86%) had latent infections. Latent infections on twigs varied markedly between years (28% in 2019 and 9.5% in 2020), while Alternaria spp. were also recovered from citrus leaves. Alternaria isolates collected during the survey were identified based on multigene sequence analysis, confirming that Alternaria alternata and Alternaria arborescens are the two species associated with infections of citrus fruits. Of the 23 isolates, 19 were identified as A. alternata and demonstrated the dominance of this species over A. arborescens. Isolates representing populations of these two species were selected as representative isolates for physiological and morphological studies. A. alternata and A. arborescens showed similar conidial dimensions but differed in the number of conidia produced. Growth rates demonstrated that A. alternata grows faster than A. arborescens at all the temperatures evaluated, except at 25 and 35 °C. The growth patterns were similar for both species. The sporulation rate of the Alternaria isolates was influenced differently by temperature. This parameter also influenced conidial germination and appressorium formation, and no significant differences were observed between Alternaria species. Pathogenicity and aggressiveness tests on detached fruit demonstrated the ability of A. alternata and A. arborescens to cause internal lesions and produce fruit drop in the orchards with no quantitative differences between them (disease severity indexes of 58 and 68%, respectively). The fungicide sensitivity tests showed that DMI fungicides are the most effective fungicides in reducing mycelial growth. The SDHI fungicides had intermediate activity against the mycelial growth but also suppressed spore germination. The spore germination assay suggested that some of the isolates included in this study might have some level of resistance to QoI and SDHI fungicides. The findings of this study provide new information about the pathogens associated with the excessive fruit drop recently observed in some California citrus groves.

Characterization of Colletotrichum Isolates Causing Colletotrichum Dieback of Citrus in California.

Dieback caused by Colletotrichum spp. is an emerging disease in California citrus groves. A large-scale survey with emphasis on seasonal variations of latent infections was conducted throughout citrus orchards in Fresno, Kern, and Tulare counties in 2019 and 2020. Latent infections on citrus leaves and twigs varied markedly between years. Isolates of Colletotrichum spp. were obtained from asymptomatic tissue, and two groups were formed based on colony and spore morphology. The morphological groups were further identified based on multigene sequence analysis using the DNA regions ITS1-5.8S-ITS2, TUB2, and GAPDH. Results revealed that isolates belong to two phylogenetic species, C. gloeosporioides and C. karstii, being C. karstii more frequently isolated. Representative isolates of each species were further selected and characterized based on the response of physiological variables to temperature. Both species had similar optimum growth temperatures but differed in maximum growth rates, with C. gloeosporioides exhibiting a greater growth rate than that of C. karstii on media. Pathogenicity tests on citrus trees demonstrated the ability of C. gloeosporioides and C. karstii to cause lesions on twigs and no differences in aggressiveness. A fungicide screening performed in this study determined that the DMI fungicides were the most effective in reducing the mycelial growth of C. gloeosporioides and C. karstii. The QoI fungicides showed a remarkably inhibitory impact on spore germination of both species. On average, C. karstii was more sensitive to the DMI fungicides than C. gloeosporioides. The findings of this study provide new information to understand the Colletotrichum dieback of citrus.

"Failure-to-Cross" in Patients Undergoing Percutaneous Peripheral Intervention: The Nonreimbursed Procedure.

BACKGROUND: Percutaneous peripheral intervention (PPI) is often the first mode of therapy for patients with symptomatic arterial occlusive disease. Technical success generally remains high although "failure-to-cross" still complicates 5-20% of cases. Extended efforts to cross long, occlusive lesions can utilize significant hospital and practitioner resources. The hospital is typically reimbursed for this effort as facility fees are charged by the hour and materials are charged per use. However, given the lack of a CPT® code for "failure-to-cross," practitioners are rarely appropriately compensated. The purpose of this study is to analyze the predictors, technical details, outcomes, and costs of "failure-to-cross" during PPI. METHODS: All PPI procedures over a 2-year period at a single institution were retrospectively reviewed. Clinical characteristics, results, costs, and reimbursements obtained from hospital cost accounting were compared among successful therapeutic interventions, crossing failures, and diagnostic angiograms without attempted intervention. RESULTS: A total of 146 consecutive PPIs were identified; the rate of "failure-to-cross" was 11.6% (17 patients). The majority of patients with "failure-to-cross" were male (82%) with single-vessel runoff (53%). Compared to successful interventions, the incidences of chronic limb-threatening ischemia (82% vs. 70%, P = 0.34) and infrapopliteal occlusive disease were similar (47% vs. 31%, P = 0.20). "Failure-to-cross" procedures were just as long as successful procedures; there were no significant differences in fluoroscopy time (27 ± 10 vs. 24 ± 14 min, P = 0.52), in-room time (106 ± 98 vs. 103 ± 44 min, P = 0.84), or contrast dye volume utilization (73 ± 37 vs. 96 ± 54 mL, P = 0.12). As expected, "failure-to-cross" procedures incurred far higher hospital charges and costs compared to noninterventional diagnostic angiograms (charges $13,311 ± 6,067 vs. $7,690 ± 1,942, P < 0.01; costs $5,289 ± 2,099 vs. $2,826 ± 1,198, P < 0.01). Despite the additional time and effort spent attempting to cross difficult lesions, the operators were reimbursed at the same low rate as a purely diagnostic procedure (average fee charge $7,360; average reimbursement $992). After 1 year, the 17 patients in whom lesions could not be crossed were treated with advanced interventional procedures with success (n = 2), surgical bypass grafting (n = 5), extremity amputation (n = 4), or no additional intervention in their salvaged limb (n = 6). CONCLUSIONS: Patients whose lesions cannot be crossed during PPI fare worse than patients undergoing successful interventions. Hospital costs and charges appropriately reflect the high technical difficulty and resource utilization of extended attempts at endovascular therapy. For practitioners, crossing lesions during PPI is truly a "pay-for-performance" procedure in that only successful procedures are reasonably reimbursed.

A Novel Topical Wound Therapy Delivery System.

INTRODUCTION: Wound care dressings have evolved over time, from bandaging to the development of occlusive dressings to negative pressure wound therapy. A novel therapeutic delivery system dressing has been cleared by the United States Food and Drug Administration. This semi occlusive wound dressing has been developed to provide local, continuous delivery of aqueous topical agents, such as therapeutics (anesthetics, antiseptics, antibiotics, steroids, topical beta-blockers, immune modulatory agents, growth factors, and fibrinolytic agents, among others), at a rate of about ¾ mL per day, thus maintaining a hydrated environment and providing topical treatment. This type of system may be beneficial in situations where systemic therapies cannot be used, wounds are small and few, wounds may need frequent application of medication or moisture, or low and steady delivery of medications is needed. OBJECTIVE: The authors assessed a delivery system dressing with different types of liquid medications for the management of hard-to-heal, chronic lower extremity wounds. MATERIALS AND METHODS: Patients aged ≥ 18 and ≤ 90 years with stalled chronic wounds > 30 days' duration were selected for the use of a topical delivery system, which consists of a semi occlusive wound dressing and fluid delivery unit that can provide local application of small therapeutic quantities of medication directly to the wound. RESULTS: Several successful cases with the use of this device are presented in which pain relief, enhancement of epithelial migration, inflammation reduction, bacterial control, and wound size reduction were achieved. CONCLUSIONS: This delivery system dressing is an effective and safe treatment option for wounds. Advantages include reduced potential of systemic side effects, flexibility in what can be delivered, constant rate of medication delivery, and convenience.

Incidencia de hernia inguinal en el Hospital Padre Billini durante el período enero 1989-1994 / Incidence of inguinal hernias at Padre Billini Hospital during the period january 1989-1994

Antecedentes: La hernia inguinal es uno de las patologías quirúrgicas mas frecuentes y nos pareció interesante determinar sus características en un hospital general. Materiales y métodos: Realizamos un estudio retrospectivo, analítico y descriptivo del 1 de enero 1989 al 31 de diciembre 1994 (6 años) en el Hospital Padre Billini, ubicado en la zona colonial de Santo Domingo, República Dominicana, revisando un total de 7432 pacientes ingresados al Servicio de Cirugía General. Resultados: Encontramos que 627 (8.4//) de los pacientes ingresados al Servicio de Cirugía General correspondía a hernias inguinales. De esos pacientes 341 (54.4//) tenían mas de 40 años de edad. El tipo de hernia mas frecuente lo constituyó la indirecta con 450 casos (71.8//). La intervención quirúrgica se realizó de forma electiva en 523 (83.4//) de los casos y de emergencia en 104 (16.6//), de las cuales 89 correspondieron a hernia encarcelada. En 3 (0.5) pacientes se presentó retención urinaria como complicación post-operatoria. En 360 casos (57.0//) se utilizó la técnica quirúrgica de Madden. La estadía promedio de los pacientes en el hospital fue de 3 días. La mortalidad fue de 0 casos (0.0//). Conclusión: un 8.4// de los pacientes ingresados al Servicio de Cirugía General en el Hospital Padre Billini padecían de hernia inguinal, patología esta que se hace mas frecuente a medida que se avanza en edad

Mesotelioma fibroso del peritoneo parietal / Fibrous esothelioma of parietal peritoneum

Se presenta el caso clínico de una paciente con mesotelioma fibroso localizado en peritoneo parietal, entidad de gran rareza en el ámbito mundial de acuerdo con las revisiones bibliográficas realizadas: en consideración a esto, se exponen los aspectos clínicos, radiológicos y endoscópicos (laparoscopia), así como la histología de la pleza resecada

Mesotelioma fibroso del peritoneo parietal / Fibrous mesothelioma of parietal peritoneum

Se presenta el caso clínico de una paciente con mesotelioma fibroso localizado en peritoneo parietal, entidad de gran rareza en el ámbito mundial de acuerdo con las revisiones bibliográficas realizadas: en consideración a esto, se exponen los aspectos clínicos, radiológicos y endoscópicos (laparoscopia), así como la histología de la pleza resecada

Correlación endoscópica-quirúrgica e histológica en las apendicitis agudas / Surgical and histologic endoscopic correlation in acute appendicitis

Se revisaron las laparoscopias de urgencia realizadas en este centro, durante un período de 4 años, se realizaron 483 laparoscopias en urgencias, se escogieron 145 diagnosticadas como positivas y presuntivas de apendicitis aguda, de éstas 141 correspondieron con los resultados obtenidos mediante la intervención quirúrgica y el estudio anatomopatológico, que representan el 97,24

de efectividad. De esto se desprenden, la gran importancia de la laparoscopia inmediata en el abdomen agudo y en especial en las supuestas apendicitis agudas

Correlación endoscópica-quirúrgica e histológica en las apendicitis agudas / Surgical and histologic endoscopic correlation in acute appendicitis

Se revisaron las laparoscopias de urgencia realizadas en este centro, durante un período de 4 años, se realizaron 483 laparoscopias en urgencias, se escogieron 145 diagnosticadas como positivas y presuntivas de apendicitis aguda, de éstas 141 correspondieron con los resultados obtenidos mediante la intervención quirúrgica y el estudio anatomopatológico, que representan el 97,24 % de efectividad. De esto se desprenden, la gran importancia de la laparoscopia inmediata en el abdomen agudo y en especial en las supuestas apendicitis agudas

Hepatitis aguda tipo B tratadas con interferon alfa leucocitario / Acute hepatitis B treatent with leukocyte alpha interferon

Se estudiaron 20 pacientes con hepatitis viral aguda (virus B) diagnosticadas por laparoscopias y biopsias, los cuales fueron distribuidos aleatoriamente en tres grupos de tratamiento, los que recibieron una dosis total de 45 millones de unidades interferón alfa por vía intraperitoneal o intramuscular entre tres y cinco días. Se compararon los resultados de4 la bilirrubina, transaminasa glutamicopirúvica y AgsHB (método ELISA), semanalmente durante el primer mes y mensualmente hasta el sexto mes. Los resultados mostraron una mayor eficacia al utilizar el tratamiento del grupo III

Hepatitis aguda tipo B tratadas con interferon alfa leucocitario / Acute hepatitis B treatment with leukocyte alpha interferon

Se estudiaron 20 pacientes con hepatitis viral aguda (virus B) diagnosticadas por laparoscopias y biopsias, los cuales fueron distribuidos aleatoriamente en tres grupos de tratamiento, los que recibieron una dosis total de 45 millones de unidades interferón alfa por vía intraperitoneal o intramuscular entre tres y cinco días. Se compararon los resultados de4 la bilirrubina, transaminasa glutamicopirúvica y AgsHB (método ELISA), semanalmente durante el primer mes y mensualmente hasta el sexto mes. Los resultados mostraron una mayor eficacia al utilizar el tratamiento del grupo III