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Soybean ( Glycine max ) allergy can be life threatening. A lack of causative immunotherapy of soybean allergy makes soybean avoidance indispensable. Detection methods are essential to verify allergen labeling and unintentional allergen cross contact during food manufacture. Here, we aimed at evaluating our previously described primers for loop-mediated isothermal amplification (LAMP) of multicopy gene ORF160b , combined with a lateral flow dipstick (LFD)-like detection, for their performance of soybean detection in complex food matrices. The results were compared with those obtained using quantitative real-time Polymerase Chain Reaction (qPCR) as the current standard of DNA-based allergen detection, and antibody-based commercial lateral flow device (LFD) as the current reference of protein-based rapid allergen detection. LAMP-LFD allowed unequivocal and reproducible detection of 10 mg/kg soybean incurred in three representative matrices (boiled sausage, chocolate, instant tomato soup), while clear visibility of positive test lines of two commercial LFD tests was between 10 and 10 2 mg/kg and depending on the matrix. Sensitivity of soybean detection in incurred food matrices, commercial retail samples, as well as various processed soybean products was comparable between LAMP-LFD and qPCR. The DNA-based LAMP-LFD proved to be a simple and low-technology soybean detection tool, showing sensitivity and specificity that is comparable or superior to the investigated commercial protein-based LFD.

The soybean ( Glycine max ) has been recognized as a frequent elicitor of food allergy worldwide. A lack of causative immunotherapy of soybean allergy makes soybean avoidance essential. Therefore, sensitive and specific methods for soybean detection are needed to allow for soybean verification in foods. Loop-mediated isothermal amplification (LAMP) represents a rapid and simple DNA-based detection method principally suitable for field-like applications or on-site analytical screening for allergens during the manufacturing of foods. This work describes the systematic development and selection of suitable LAMP primers based on soybean multicopy genes. The chemistry applied allows for a versatile detection of amplified DNA, using either gel electrophoresis, fluorescence recording, or a simple Lateral Flow Dipstick (LFD). LAMP based on the ORF160b gene was highly specific for the soybean and may allow for a detection level equivalent to approximately 10 mg soy per kg food. Various soybean cultivars were detectable at a comparable level of sensitivity. LAMP combined with LFD-like detection facilitates a simple, highly specific and sensitive detection of the soybean without the need for expensive analytical equipment. In contrast to the majority of antibody-based methods for soybean detection, all identified primer sequences and optimized protocols are disclosed and broadly available to the community.

We retrospectively analyzed a cohort of 109 subjects treated for recurrent Clostridium difficile infection with fecal microbiota transplantation (FMT) at a tertiary referral center between 2011 and 2014 to determine risk factors for FMT failure. In a multivariate analysis, failure to use an oral vancomycin taper preceding FMT was associated with a significant risk of FMT failure (odds ratio, 0.15; 95% confidence interval, .007-.40).

In summer 2015, three unrelated solid organ transplant recipients in Phoenix, Arizona, had meningoencephalitis suggestive of West Nile virus (WNV) infection. Testing was inconclusive but was later confirmed as St. Louis encephalitis (SLE). We retrospectively reviewed clinical manifestations, treatment, and outcomes of these transplant recipients. Common symptoms were fever, rigors, diarrhea, headache, and confusion. One patient died 3 days after hospitalization. Therapy for the other two patients was initiated with interferon α-2b (IFN) and intravenous IgG (IVIG; IFN plus IVIG in combination). Both patients tested positive for WNV by serologic assay, but SLE virus (SLEV) infection was later confirmed by plaque reduction neutralization test at a reference laboratory. Clinical improvement was observed within 72 h after initiation of IFN plus IVIG. SLEV has been an uncommon cause of neuroinvasive disease in the United States. Accurate, timely diagnosis is hindered because of clinical presentation similar to neuroinvasive WNV and SLE, serologic cross-reactivity, and lack of a commercially available serologic assay for SLEV. There is currently no approved therapy for flaviviral neuroinvasive disease. Anecdotal reports indicate varying success with IFN, IVIG, or IFN plus IVIG in WNV neuroinvasive disease. The same regimen might be of value for immunocompromised persons with neuroinvasive SLEV infection., (© 2017 The American Society of Transplantation and the American Society of Transplant Surgeons.)

Acticoat™ has antimicrobial and anti-inflammatory effects which aid wound healing. However, in vitro studies indicate that Acticoat™ is cytotoxic and clinical and in vivo studies suggest that it may delay healing in acute wounds. Therefore, this study investigated the effects of Acticoat™ on healing in acute full-thickness excisional wounds. Using a porcine model, healing was assessed on days 3, 6, 9 and 15 post-wounding. Five wounds dressed with Acticoat™ and five wounds dressed with polyurethane film (control) were assessed per day (n = 40 wounds). The rate of healing, inflammatory response, restoration of the epithelium and blood vessel and collagen formation were evaluated. No difference was found in the rate of healing between wounds treated with Acticoat™ and the control wounds. Inflammation was increased in Acticoat™-treated wounds on day 3 post-wounding compared to the control wounds. However, by day 15 post-wounding, the epithelium of the Acticoat™-treated wounds closely resembled normal epithelium. Acticoat™-treated wounds also contained a higher proportion of mature blood vessels, and differences in collagen deposition were apparent. Despite inducing an inflammatory response, Acticoat™ did not delay healing in acute wounds. Conversely, the improved quality of the epithelium and blood vessels within Acticoat™-treated wounds indicates that Acticoat™ has a beneficial effect on healing., (© 2015 Medicalhelplines.com Inc and John Wiley & Sons Ltd.)

Introduction: Coccidioidomycosis is an endemic fungal infection caused by the soil-dwelling fungi, Coccidioides species. Coccidioidal infections may be asymptomatic in up to two-thirds of infected persons. Pulmonary coccidioidomycosis is the most common form of symptomatic infection. Fluconazole is the antifungal agent typically used to treat pulmonary coccidioidomycosis. Other azoles and amphotericin B products may be prescribed to treat nuanced aspects of coccidioidomycosis., Areas Covered: This review discusses current literature regarding medical treatment options, including the various triazoles and amphotericin B products. In addition, we discuss uncomplicated and complicated pulmonary infections and their sequelae and the approach to managing coccidioidomycosis in certain populations of patients, such as pregnant women, transplant recipients, individuals infected with human immunodeficiency virus, and recipients of tumor necrosis factor-α inhibitors. Expert commentary: Symptomatic coccidioidomycosis can present physicians with a number of challenges, including the lack of sensitivity and specificity of diagnostic tests and lack of a standard treatment approach for all patients with the infection. Ongoing and future clinical trials will determine the optimal diagnostic, therapeutic, and prophylactic approaches, particularly for patients with comorbid conditions.

Colonoscopy is the standard technique in the diagnosis and treatment of colorectal neoplasia, but small adenomas and even advanced lesions can be missed during the procedure. With large scale screening colonoscopy programs installed, information on quality of colonoscopy in primary care is essential, but scarcely available. Over a period of 45 months, we prospectively included all those patients in our study, who underwent major colonic surgery at our institution and who had undergone a colonoscopy within 42 days prior to the operation. 89 men and 100 women, median age 71 years, were included. The majority of these operations were performed for colorectal carcinoma (125), other malignant tumors (4), suspected malignancies (6) or large adenomas (14). The pathologist inspected the resected colonic segment, and we compared his findings with the colonoscopy report. Colonoscopies had been performed by 22 doctors in 13 institutions. Median length of the resected colonic segments was 20 cm (range 3 to 135 cm), total length was 41,21 metres. In 14 segments the pathologist identified 28 neoplastic lesions not described in the endoscopy report. Colonoscopy had missed 2 carcinomas, both in the right colon, and a 12 mm tubulo-villous adenoma with high-grade dysplasia. Another 25 tubular adenomas had been missed, 2 measuring 10 mm, 7 between 5 and 9 mm and 16 smaller than 5 mm. We conclude that primary care colonoscopy misses neoplastic lesions in a significant number of procedures. Most of the missed lesions in our high risk group of patients would have been of little clinical consequence. In a small, but clinically important number of cases, however, advanced adenomas and even colorectal carcinomas were missed by endoscopy., (© Georg Thieme Verlag KG Stuttgart · New York.)

Prion protein (PrP(C)) has neuroprotective functions and herein we demonstrate that astrocytes from PrP(C)-over-expressing mice are more resistant to induced cell death than wild-type astrocytes. The Stress-Inducible-Protein 1 (STI1), a PrP(C) ligand, prevents cell death in both wild-type and PrP(C)-over-expressing astrocytes through the activation of protein-kinase-A. Cultured embryonic astrocytes and brain extracts from PrP(C)-over-expressing mice show higher glial fibrillary acidic protein expression and reduced vimentin and nestin levels when compared to wild-type astrocytes, suggesting faster astrocyte maturation in the former mice. Our data indicate that PrP(C) levels modulate astrocyte development, and that PrP(C)-STI1 interaction contributes to protect against astrocyte death., (Copyright © 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.)

beta 2 microglobulin knockout (beta2M-/-) mice lack CD8+ T and natural killer T cells. We hypothesized that beta 2M-/- mice are resistant to lethal intraabdominal sepsis. To test this hypothesis, mortality, cytokine production, and physiologic function were assessed in beta 2M-/- mice during sepsis caused by cecal ligation and puncture (CLP). beta 2M-/- mice survived significantly longer than wild-type mice after CLP but ultimately exhibited 100% mortality. Treatment of beta 2M-/- mice with anti-asialoGM1 to deplete natural killer cells conferred greater than 70% long-term survival. Compared with wild-type mice, beta 2M-/- mice treated with anti-asialoGM1 produced decreased amounts of proinflammatory cytokines and did not exhibit hypothermia or metabolic acidosis after CLP. Adoptive transfer of CD8+ T and natural killer cells into beta 2M-/- mice treated with anti-asialoGM1 re-established CLP-induced mortality. CD8 knockout mice treated with anti-asialoGM1, which are specifically deficient in CD8+ T and natural killer cells, exhibited 40% long-term survival after CLP. Furthermore, treatment of wild-type mice with antibodies to CD8 and asialoGM1 conferred a significant survival benefit compared with wild-type mice treated with nonspecific IgG. These findings demonstrate that beta 2M-/- mice treated with anti-asialoGM1 are resistant to CLP-induced mortality and that depletion of CD8+ T and natural killer cells largely accounts for the survival benefit observed in these mice.

The 72 mesothelioma-like tumors of the pleura (MLTP) found among 33 500 autopsy cases collected over more than 30 years are reviewed. MLTP have a worse prognosis than the 106 cases of pleural mesothelioma autopsied in our institutes with regard to survival time and metastatic spread. In MLTP, adenocarcinomas predominate with a wide range of histological and cytological variation and prominent development of connective tissue having its origin in the periphery of the lung. These intrapulmonary primary tumors often fulfill the criteria of pulmonary scar cancer. Etiologically, there is no correlation between the origin of this tumor and smoking or exposure to asbestos. The absence of mucus formation and glandular differentiation, together with the presence of spindle-shaped carcinoma components and strong mesothelial or stroma proliferation, can make the differential diagnosis between this tumor type and mesothelioma difficult. Immunohistological investigations were performed on 11 cases with antibodies against intermediate filament proteins, vascular endothelium, collagen IV, macrophage antigens, carcinoembryonic antigen (CEA), LeuM1, and the antibody BerEP4. Our investigation shows that a battery of several tumor markers, such as antibodies against LeuM1, CEA, and the antibody BerEP4, as well as staining with periodic acid/Schiff/diastase discriminate primary from secondary pleural neoplasms, whilst intermediate filament proteins alone are of little diagnostic value.

Seven adrenal carcinomas and seventeen pheochromocytomas (PHs), two of which were clinically associated with a Cushing's syndrome and one associated with multiple endocrine neoplasia Type II (MEN-II), were investigated immunohistologically with a panel of antibodies against intermediate filament proteins, a proliferation-associated nuclear antigen (Ki-67), neuroendocrine tumor markers, and different hormones on paraffin-embedded tissue sections and, from 19 cases, also on frozen tissue sections. Synaptophysin proved to be the most reliable tumor cell marker on both snap-frozen and paraffin-embedded tissue but, like antibodies against NSE, yielded unspecific stainings in the carcinomas. The two Cushing-associated pheochromocytomas (CaPH) showed the same immunohistological profile as the other PHs, except one chromogranin-negative tumor. Five PHs showed weak reactivity for calcitonin, one for serotonin, and two for a-HCG in small amounts. All PHs lacked other hormone expression, including ACTH. The average growth fraction was small (2.2%) in 13 cases, but 80% of the tumor cells were proliferating in one case of CaPH. Adrenal carcinomas showed only weak or no expression of keratin in one case, a homogenous or droplet, non-filamentous cytoplasmic staining with antibodies against neurofilament in frozen tissue section, and they were completely chromogranin-negative. The average growth fraction was 7.6% in 5 cases.

In addition to diagnosing deep-vein thrombosis of the lower limb and pelvis, determination of the age of the thrombus is of considerable importance in guiding treatment. The present study was done to find out whether thrombus age can be determined by ultrasound morphology. 138 examinations were carried out on 76 lower limbs with phlebographically demonstrated thrombosed veins. Diameter of the thrombosed vein, echogenicity of the thrombus and demarcation of the venous wall from its surroundings were determined by colour-coded duplex sonography. The results were compared with histological evidence or with the age of the thrombus as determined by history and clinical course. The diameter of the thrombosed vein in conjunction with the history and clinical data makes it possible to determine the age of a thrombus. Echogenicity of the thrombus and demarcation of the venous wall do not provide any reliable information on thrombus age.

The Epstein-Barr virus (EBV) is consistently associated with undifferentiated nasopharyngeal carcinoma (NPC). There is, however, conflicting evidence as to whether squamous cell NPCs are also EBV-associated. Moreover, it has been proposed that other epithelial tumours, particularly thymomas and thymic carcinomas, should be included in the group of EBV-associated neoplasias. However, since the viral DNA in these studies was demonstrated only in extracted DNA, the cellular origin of the viral DNA is uncertain. We have therefore investigated 152 epithelial tumours from various sites for the presence of EBV-DNA by in situ hybridization with 35S-labelled probes. Sixty-eight of 77 undifferentiated NPCs showed an EBV-specific autoradiographic signal, thus confirming the strong association of this tumour type with EBV even in geographical areas where undifferentiated NPC is not endemic. None of eight squamous cell NPCs showed an EBV-specific signal. All of 15 carcinomas with a similar morphology to undifferentiated NPC but from different anatomic sites (thymus, tonsil, breast) were EBV-negative as were 9 thymomas, 26 squamous cell carcinomas of the palatine tonsil, and 14 cervical carcinomas. Our results therefore suggest a unique association of EBV with undifferentiated NPC and support concepts assigning different biological properties to undifferentiated NPC as compared with squamous cell NPC.

In a prospective study, 188 patients with suspected deep venous thrombosis were examined by color-coded duplex sonography. In 114 patients the diagnosis of deep venous thrombosis was based on the criteria of 1) compressibility, 2) blood flow changes during compression, and 3) pelvic level during Valsalva maneuver. In all patients the results were compared with data from contrast venography. Venography was performed by an investigator, who was blinded to the prior findings. Sensitivity and specificity of the color-coded duplex sonography in diagnosing deep vein thrombosis were 96% and 97%, respectively. In the other 74 patients, in which the diagnosis of deep vein thrombosis was made by phlebography and/or ultrasound, a follow-up by color-coded duplex sonography was performed to determine the age of thrombosis. Criteria considered to determine the thrombus age included 1) margin of the vessel, 2) echogenicity; and 3) venous diameter. In 20 patients the results were compared to the age of thrombus that had been determined by histologic criteria (group I). In 54 patients the results were compared with the age that had been determined by patient history and symptoms (group II). A significant correlation was found between the age of thrombosis and the venous diameter (p less than 0.001). When thrombosis was less than 10 days old, the venous diameter was at least twice that of the diameter of the accompanying artery. The two other criteria, echogenicity and margin of the venous wall, however, did not prove to be accurate criteria to determine the age of deep venous thrombosis.

The nature of the atypical, often multinucleated stromal cells proliferating in the so-called fibroepithelial polyps of the vagina (FEPV) is obscure. The authors therefore investigated immunohistochemically four polyps of this type with a panel of monoclonal and polyclonal antibodies, including those directed against intermediate filaments, steroid hormone receptors, and macrophage antigens. The authors found that the atypical stromal cells of FEPV expressed vimentin, desmin, and receptors for estrogen and progesterone but lacked all the other markers studied. The homogenous antigen profile strongly suggests that FEPV form a disease entity whose characteristic cells are special hormone-sensitive stromal cells with a myoid component of differentiation. Morphologically similar stromal cells expressing steroid hormone receptors but lacking desmin were found in myxomatoid subepithelial regions of the vaginal wall and the cervix.

In situ hybridization (ISH) using 35S-labeled probes was applied to the detection of viral DNAs in 115 epithelial tumours from various sites. Epstein-Barr virus (EBV) DNA was detected in 83% of undifferentiated nasopharyngeal carcinomas (NPC) while none of seven squamous cell NPC displayed an EBV-specific autoradiographic signal. Also, all other tumours investigated, including thymomas, thymic carcinomas, tonsillar carcinomas and medullary breast carcinomas were negative upon ISH to EBV-specific probes. These results suggest a unique association of undifferentiated NPC with EBV. 28 tonsillar carcinomas were further analyzed for the presence of human papillomavirus (HPV) DNA. Six carcinomas showed an HPV16-specific signal while hybridization to HPV6- and HPV11-specific probes yielded negative results. No HPV-DNA was detected in any of 30 tonsils with chronic inflammation, thus excluding that the finding of HPV16 in a proportion of tonsillar carcinomas is merely circumstantial. These results point to a possible etiologic role of HPV16 in the pathogenesis of some tonsillar carcinomas.

Among 54 mediastinal tumours we examined in the past 20 years, there were 5 cases of primary thymic carcinomas, each with widespread metastases. Histological features in three cases were consistent with lymphoepithelioma-like carcinoma. One case showed an epidermoid pattern with keratotic pearls resembling Hassall bodies. One undifferentiated carcinoma developed from a cortical thymoma. Epstein-Barr virus could not be detected in tumour tissue with in situ hybridization. A review of the literature revealed only 94 well-documented cases of thymic carcinoma. Both thymic carcinomas and thymomas are neoplasms of the thymic epithelial cells, but thymic carcinomas are obviously histologically malignant and usually not associated with any parathymic syndromes. Epidermoid and lymphoepithelioma-like carcinomas are described along with special forms, such as small- and clear-cell carcinomas, basaloid, sarcomatoid, mucoepidermoid, and adenocystic carcinoma. Compared to the other forms, lymphoepithelioma-like carcinoma has a poor prognosis in regard to metastases and rate of survival. Some thymic carcinomas may develop from pre-existing thymomas.

Cryosurgical treatment was applied to normal liver tissue of 13 clinically intact rats, with a view to investigating effects of freezing temperatures upon normal liver tissue. The animals were sacrificed after different intervals from surgery, and their liver tissue was macroscopically and histologically examined. Major postoperative complications were not observed but for one animal which died of wound healing disorder. The method seems to be suitable also for treatment of liver cell carcinoma or liver metastases.

Three cases of pulmonary blastoma (PB) were investigated microscopically with conventional stainings and immunohistochemically with monoclonal antibodies to cytokeratin, vimentin, desmin and neurofilament protein. The tumors differed in terms of morphology as well as of immunohistochemistry. Two were epithelial and mesenchymal mixed tumors, and the remaining one was a monophasic tumor of a typical blastemic character. The two mixed tumors also differed from each other. In one of them, the epithelial and mesenchymal component expressed cytokeratin and vimentin in a clear-cut manner without any transition. The other mixed tumor displayed a gradual epithelial-to-mesenchymal transition accompanied by a switch in the expression of cytokeratin and vimentin. The third tumor was of pure mesenchymal origin, expressing vimentin in the majority of cells and desmin in few cells. It is concluded that the PB is a morphologically and histogenetically heterogeneous tumor. Metaplastic changes may take place within a PB and make the recognition of embryogenesis more difficult.

Cryosurgical treatment was applied to solid liver tumours of rats with a view to studying the effects of freezing temperatures upon liver carcinoma. Such treatment was applied to twelve rats to which carcinomas, 0.5 cm to 3.0 cm in size, had been implanted. Cryosurgical treatment was successful in one case. Many of the animals died of complications which resulted from freezing of surrounding organs, such as stomach or greater liver vessels.

In 100 pleural mesotheliomas follow up x-ray studies were compared to autopsy findings. Initially x-ray examinations show effusions in 62%, diffuse pleural thickening in 29% and solitary pleural nodules in 6%, prior to death effusions and pleural thickening are usually combined. In 17 tumor nodules doubling times ranged from 0.5 to 3 months. The epithelial type of mesotheliomas is predominantly combined with pleural effusions, while pleural thickening is more often found in the mesenchymal type of the tumor.

Different basic mycological observations were made in relation to a fatal case of aspergilloma of the lung in a 45-year-old male. Cultural examination of the sputum and autopsy material from the lung revealed the predominant presence of a white fungus along with a scarce growth of the typical Aspergillus fumigatus. The white isolate produced conidial heads almost similar to that of A. fumigatus but with apparent septation in the phialides. With the help of the immunodiffusion test and use of the antigens from the typical A. fumigatus strain and from the white isolate, antibodies could be detected in the serum of this patient and an antigenetic relationship between these fungi could be observed. It is assumed that this white isolate is a variant of A. fumigatus and its presence in the lung is an indication of chronic or progressed aspergillosis caused by A. fumigatus. Cultural characteristics and developmental morphology of this fungus and the serodiagnostic value of the antigen derived from it will be described in detail in separate communications.

An autopsy case of a 45-year-old man is presented. The man had sustained a minor head injury in a traffic accident 11 years ago previously and 5 years later had a cerebral infarction due to thrombosis of the carotid artery. There was histological evidence of a traumatic lesion of the vessel wall. Several contradictory expert opinions were rendered regarding the questionable causal relationship between occlusion of the artery and the previous trauma. In the court of appeal, an agreement was arrived at between the widow of the deceased and the third-party insurance of the person responsible for the accident.