Paracellular intestinal permeability of chickens induced by DON and/or C. jejuni is associated with alterations in tight junction mRNA expression.

Toxins, antigens, and harmful pathogens continuously challenge the intestinal mucosa. Therefore, regulation of the intestinal barrier is crucial for the maintenance of mucosal homeostasis and gut health. Intercellular complexes, namely, tight junctions (TJs), regulate paracellular permeability. TJs are mainly composed of claudins (CLDN), occludin (OCLN), tight junction associated MARVEL-domain proteins (TAMPS), the scaffolding zonula occludens (ZO) proteins and junction-adhesion molecules (JAMs). Different studies have shown that a Campylobacter infection can lead to a phenomenon so-called "leaky gut", including the translocation of luminal bacteria to the underlying tissue and internal organs. Based on the effects of C. jejuni on the chicken gut, we hypothesize that impacts on TJ proteins play a crucial role in the destructive effects of the intestinal barrier. Likewise, the mycotoxin deoxynivalenol (DON) can also alter gut permeability in chickens. Albeit DON and C. jejuni are widely distributed, no data are available on their effect on the tight junctions' barrier in the broiler intestine and consequences for permeability. Therefore, the aim of this study was to analyze the interaction between DON and C. jejuni on the gut barrier by linking permeability with gene expression of TJ proteins and to determine the relationships between the measurements. Following oral infection of birds with C. jejuni NCTC 12744 at 14 days of age, we demonstrate that the co-exposure with DON has considerable consequences on gut permeability as well as on gut TJ mRNA expression. Co-exposure of DON and C. jejuni enhanced the negative effect on paracellular permeability of the intestine, which was also noticed for the bacteria or the mycotoxin alone by the Ussing chamber technique at certain time points in both jejunum and caecum. Furthermore, the increased paracellular permeability was associated with significant changes in TJ mRNA expression in the small and large intestine. The actual study demonstrates that co-exposure of broiler chickens to DON and C. jejuni resulted in a decreased barrier function via up-regulation of pore-forming tight junctions (CLDN7 and CLDN10), as well as the cytosolic TJ protein occludin (OCLN) that can shift to various paracellular locations and are therefore able to alter the epithelial permeability. These findings indicate that the co-exposure of broiler chickens to DON and C. jejuni affects the paracellular permeability of the gut by altering the tight junction proteins. Furthermore, analysing of correlations between TJs revealed that the mRNA expression levels of most tight junctions were correlated with each other in both jejunum and caecum. Finally, the findings indicate that the molecular composition of tight junctions can be used as a marker for gut health and integrity.

Establishment of a novel probe-based RT-qPCR approach for detection and quantification of tight junctions reveals age-related changes in the gut barriers of broiler chickens.

Tight junctions (TJs) play a dominant role in gut barrier formation, therefore, resolving the structures of TJs in any animal species is crucial but of major importance in fast growing broilers. They are regulated in molecular composition, ultrastructure and function by intracellular proteins and the cytoskeleton. TJ proteins are classified according to their function into barrier-forming, scaffolding and pore-forming types with deductible consequences for permeability. In spite of their importance for gut health and its integrity limited studies have investigated the TJs in chickens, including the comprehensive evaluation of TJs molecular composition and function in the chicken gut. In the actual study sequence-specific probes to target different TJ genes (claudin 1, 3, 5, 7, 10, 19, zonula occludens 1 (ZO1), occludin (OCLN) and tricellulin (MD2)) were designed and probe-based RT-qPCRs were newly developed. Claudin (CLDN) 1, 5, ZO1 and CLDN 3, 7, MD2 were engulfed in multiplex RT-qPCRs, minimizing the number of separate reactions and enabling robust testing of many samples. All RT-qPCRs were standardized for chicken jejunum and caecum samples, which enabled specific detection and quantification of the gene expression. Furthermore, the newly established protocols were used to investigate the age developmental changes in the TJs of broiler chickens from 1-35 days of age in the same organ samples. Results revealed a significant increase in mRNA expression between 14 and 21days of age of all tested TJs in jejunum. However, in caecum, mRNA expression of some TJs decreased after 1 day of age whereas some TJs mRNA remained constant till 35 days of age. Taken together, determining the segment-specific changes in the expression of TJ- proteins by RT-qPCR provides a deeper understanding of the molecular mechanisms underpinning pathophysiological changes in the gut of broiler chickens with various etiologies.

Efficacy of Radiation Therapy for the Treatment of Sialocele in Dogs.

BACKGROUND: Sialocele is a collection of saliva that has leaked from a damaged salivary gland or duct and is surrounded by granulation tissue. Surgery is the recognized first-line treatment. Recurrence rate after surgery is 5-14%. Salivary gland tissue is very sensitive to radiation therapy (RT). HYPOTHESIS/OBJECTIVES: Radiation therapy will be useful for the treatment of sialocele. The aims were to characterize response rate and clinical course of dogs with sialocele treated with RT and to determine a starting dose for clinical use. ANIMALS: Eleven dogs with sialocele. METHODS: Retrospective study of response and outcome after RT. RESULTS: All dogs had cervical sialocele. Seven dogs (63.6%) were treated with 3 weekly fractions of 4 Gray (Gy); (total dose, 12 Gy). Three dogs (27.3%) received 4 fractions of 4 Gy (16 Gy) and 1 dog received 5 fractions of 4 Gy (20 Gy) on a Monday-Wednesday-Friday schedule. Six dogs (54%) achieved a complete response (CR), and 5 dogs (45%) achieved a partial response (PR). Three dogs had progression of their sialocele 2, 3, and 9 months after RT; all three had received 12 Gy initially and 2 received 2 additional fractions of 4 Gy (cumulative total dose, 20 Gy) and subsequently achieved remission for >2 years. CONCLUSIONS AND CLINICAL IMPORTANCE: Radiation therapy is useful for the treatment of recurrent sialocele refractory to surgical management and a minimum total dose of 16 or 20 Gy in 4 Gy fractions appears effective.

The effect of hearing loss on source-distance dependent speech intelligibility in rooms.

Westermann and Buchholz [(2015). J. Acoust. Soc. Am. 137(2), 757-767] found substantial improvements in speech reception thresholds (SRTs) for normal hearing listeners in a reverberant auditorium when the target talker was separated in distance from a two-talker masker. This study applied similar methodology, but tested listeners with a hearing impairment. On average, the participants received a 7 dB benefit in SRTs when the target was fixed at 0.5 m and the masker was moved from 0.5 to 10 m. But when the target was moved away, the SRTs increased by 5 dB. This indicates that hearing impaired listeners have difficulties suppressing nearby maskers while focusing attention on a far target.

Oxo-bridge scenario behind single-site water-oxidation catalysts.

High-oxidation-state decay of mononuclear complexes [RuTB(H2O)](2+) (X(2+), where B = 2,2'-bpy or bpy for X = 1; B = 5,5'-F2-bpy for X = 2; B = 6,6'-F2-bpy for X = 3; T = 2,2':6',2″-terpyridine) oxidized with a large excess of Ce(IV) generates a manifold of polynuclear oxo-bridged complexes. These include the following complexes: (a) dinuclear [TB-Ru(IV)-O-Ru(IV)-(T)(O)OH2](2+) (1-dn(4+)), [TB-Ru(III)-O-Ru(III)-T(MeCN)2](4+) (1-dn-N(4+)), and {[Ru(III)(trpy)(bpy)]2(µ-O)}(4+) (1-dm(4+)); (b) trinuclear {[Ru(III)(trpy)(bpy)(µ-O)]2Ru(IV)(trpy)(H2O)}(ClO4)5(6+) (1-tr(6+)) and {[Ru(III)(trpy)(bpy)(µ-O)]2Ru(IV)(pic)2}(ClO4)4 (1-tr-P(4+), where P is the 2-pyridinecarboxylate anion); and (c) tetranuclear [TB-Ru(III)-O-TRu(IV)(H2O)-O-TRu(IV)(H2O)-O-Ru(III)-TB](8+) (1-tn(8+)), [TB-Ru(III)-O-TRu(IV)(AcO)-O-TRu(IV)(AcO)-O-Ru(III)-TB](6+) (1-tn-Ac(6+)), and [TB-Ru(II)-O-TRu(IV)(MeCN)-O-TRu(IV)(MeCN)-O-Ru(II)-TB](6+) (1-tn-N(6+)). These complexes have been characterized structurally by single-crystal X-ray diffraction analysis, and their structural properties were correlated with their electronic structures. Dinuclear complex 1-dm(4+) has been further characterized by spectroscopic and electrochemical techniques. Addition of excess Ce(IV) to 1-dm(4+) generates dioxygen in a catalytic manner. However, resonance Raman spectroscopy points to the in situ formation of 1-dn(4+) as the active species.

Small bite-angle P-OP ligands for asymmetric hydroformylation and hydrogenation.

A series of small bite-angle phosphine-phosphite (P-OP) ligands have been synthesized by a two-step method. The key intermediate was prepared by an unprecedented asymmetric carbonyl reduction of a phosphamide using the CBS (Corey-Bakshi-Shibata) catalyst. The topology of these ligands (a configurationally stable stereogenic carbon with two heteroatom substituents) and their small bite-angle (created by the close proximity of the two ligating groups to the metal center) together provide a rigid asymmetric environment around this center, enabling high stereoselectivity in hydroformylations and hydrogenations of standard substrates.

Mononuclear ruthenium-water oxidation catalysts: discerning between electronic and hydrogen-bonding effects.

New mononuculear complexes of the general formula [Ru(trpy)(n,n'-F2-bpy)X](m+) [n = n' = 5, X = Cl (3(+)) and H2O (5(2+)); n = n' = 6, X = Cl (4(+)) and H2O (6(2+)); trpy is 2,2':6':2"-terpyridine] have been prepared and thoroughly characterized. The 5,5'- and 6,6'-F2-bpy ligands allow one to exert a remote electronic perturbation to the ruthenium metal center, which affects the combination of species involved in the catalytic cycle. Additionally, 6,6'-F2-bpy also allows through-space interaction with the Ru-O moiety of the complex via hydrogen interaction, which also affects the stability of the different species involved in the catalytic cycle. The combination of both effects has a strong impact on the kinetics of the catalytic process, as observed through manometric monitoring.

[Combined use of irradiation and DNA tumor vaccine to treat canine oral malignant melanoma: a pilot study]. / Kombinierte Anwendung von Strahlentherapie und DNS Tumor Vakzine zur Behandlung des oralen malignen Melanoms beim Hund: Eine Pilotstudie.

Melanoma is the most common oral tumor in dogs, characterized by rapid growth, local invasion, and high metastatic rate. The goal of this study was to evaluate the combination of radiation therapy and DNA tumor vaccine. We hypothesized, that the concurrent use would not increase toxicity. Nine dogs with oral melanoma were treated with 4 fractions of 8 Gray at 7-day intervals. The vaccine was given 4 times every 14 days, beginning at the first radiation fraction. Local acute radiation toxicities were assessed according to the VRTOG toxicity scoring scheme over a time period of 7 weeks. In none of the evaluated dogs, mucositis, dermatitis and conjunctivitis exceeded grade 2. In 3 dogs mild fever, lethargy, and local swelling at the injection site were seen after vaccine application. In conclusion, the concurrent administration of radiation therapy and vaccine was well tolerated in all dogs.

Multimodal therapeutic approach and interdisciplinary challenge for the treatment of unresectable head and neck squamous cell carcinoma in six cats: a pilot study.

Feline head and neck squamous cell carcinoma (SCC) is a loco-regional disease harbouring a poor prognosis. The complex anatomic location precludes aggressive surgical resection and tumours recur within weeks to few months. Response to chemotherapy and local control after radiation therapy has been disappointing. In this study, a multimodal approach including medical treatment (thalidomide, piroxicam and bleomycin), radiation therapy (accelerated, hypofractionated protocol) and surgery was attempted in six cats. Treatment was well tolerated. Three cats with sublingual SCC were alive and in complete remission at data analysis closure after 759, 458 and 362 days. One cat with laryngeal SCC died of renal lymphoma after 51 days and the other with maxillary SCC died of a primary lung tumour 82 days after diagnosis. In both cats, the SCC was in complete remission. Only one cat developed metastases after 144 days. These encouraging preliminary results merit further evaluation in future trials.

Note: multiplexed multiple-tau auto- and cross-correlators on a single field programmable gate array.

We introduce a new multiple-tau hardware correlator design for computing fluorescence correlation functions (CFs) in real time. Use of hardware resources is minimized by scheduling the computation of different segments of the CFs on a single correlator block. Simultaneous calculation of two multiple-tau autocorrelation (ACFs) and two cross-correlation functions (CCFs) is implemented in LabVIEW on a National Instruments field programmable gate array (FPGA) card with a minimal sampling time of 400 ns. Raw data are stored with a time resolution of 50 ns. The design can be easily adapted to other FPGA cards and extended to more inputs.

[New concepts in human oncology: is it possible to use them in veterinary medicine as well?]. / Neue Konzepte in der humanen Onkologie: Können sie in der Veterinärmedizin eingesetzt werden?

In human oncology, novel targeted therapy focusing on monoclonal antibodies and tyrosine kinase inhibitors has become an attractive anticancer strategy. The introduction of antiangiogenetic drugs and metronomic chemotherapy has also increased the therapeutic arsenal. Chemotherapy still plays a key role in the treatment of many tumors affecting dogs and cats. However, novel anticancer strategies (including tyrosine kinase inhibitors and monoclonal antibodies, as well as antiangiogenetic treatments) are becoming relevant in veterinary medicine, too. The goal of this review is to describe new therapeutic strategies for cancer treatment in veterinary medicine, including less well-known chemotherapeutic drugs.

Allosteric P═O-based receptors for dicarboxylic acids.

The synthesis of new P═O-disubstituted receptors with appended crown ethers and their properties as receptors for dicarboxylic acids have been studied. High affinities have been observed (oxalic and malonic acids with 4-, 5-, 6-, or 8-crown ethers). Binding of a cationic effector within the crown ether unit resulted in a positive "allosteric" effect, which has been determined to be K(rel) = 7 in the best case (binding of malonic acid with Li(+) @ rac-3b).

[A new total knee arthroplasty with physiologically shaped surfaces. Part 2: First clinical results]. / Eine neuartige Kniegelenksendoprothese mit physiologischer Gelenkform. Teil 2: Erste klinische Ergebnisse.

The human medial tibial plateau is concave, whereas the lateral tibial plateau is convex. In a normal knee, the convex femoral condyles roll and glide on the tibia during the standing phase of walking. The designs of most commercially available knee prostheses do not take this morphological feature into consideration. The novel design of the AEQUOS G1 knee replacement prosthesis is based on the natural anatomy of the knee joint, with a convex lateral tibia plateau and a sagittal offset of the medial and lateral compartments. Following extensive development and testing, initial clinical results of the AEQUOS G1 prosthesis in a mulitcenter study are presented. From Mai 2005 to March 2007, 158 patients in 4 clinics underwent total knee arthroplasty with the AEQUOS G1 and agreed to participate in the study. Patients were evaluated preoperatively and at 3, 6 and 12 months of follow-up using a standardized protocol that included the American Knee Society Score (AKSS), the Oxford Knee Score (OKS) and the Visual Analog Scale (VAS) for pain. After 3 months, 151 patients appeared for follow up appointments, after 6 months, 134, and after 12 months, 127. The mean range of motion preoperatively was 97.0 degrees (+/-19.9 degrees ) and 107.5 degrees (+/-15.9 degrees ) 12 months after surgery. The AKSS, as well as the modified OKS, significantly improved (p<0.0001) from preoperative scores of 98.8 (+/-35.8) and 37.3 (+/-6.9) points, respectively, to 165.8 (+/-34.1) and 21.9 (+/-7.8) points, preoperatively, and 12 months postoperatively. The VAS score significantly decreased (p<0.001) from 7.4 (+/-1.8) points preoperatively to 1.9 (+/-2.2) points 12 months postoperatively.One implant was revised because of arthrofibrosis and another due to patellar luxation. Two patients required revision because their implants revealed malalignement with ligamentous instability. No infections, aseptic loosening or other implant-specific complications were observed at this early follow-up. Good clinical results were observed at early follow-up with the AEQUOS G1 knee arthroplasty. However, longer follow-up is necessary for a general evaluation of the implant.

Primary pulmonary spindle cell tumour (haemangiopericytoma) in a dog.

Haemangiopericytoma is a soft tissue sarcoma believed to originate from pericytes. These tumours are commonly located on the skin and subcutaneous tissue of dogs and are most commonly found on the limbs. To the authors' knowledge, primary lung haemangiopericytomas have not been previously described in dogs. This case report describes the diagnostic evaluation and treatment of a primary haemangiopericytoma of the lung in a 10-year-old male, neutered, Siberian husky dog. Staging of the tumour was performed using a computed tomography scan of the thorax and a computed tomography-guided fine-needle aspiration biopsy of the lesion. Treatment was a right caudal lobectomy from a right lateral approach. No regional lymph node changes were noted on computed tomography or intraoperative assessments. Histopathology confirmed a spindle cell tumour that stained positive for vimentin and negative for desmin and S-100.

Dioxirane mediated asymmetric epoxidations: stereochemical studies via isotopic labeling.

A study, via isotopic labeling, of the stereoselective processes in a Shi-type epoxidation, has revealed that the chiral platform provided by the catalyst mediates the transfer of the pro-S "O" of the related dioxirane species to the alkene in a doubly stereoselective manner.

Identification of proximal 1p36 deletions using array-CGH: a possible new syndrome.

Monosomy 1p36 is the most common terminal deletion syndrome with an estimated occurrence of 1:5000 live births. Typically, the deletions span <10 Mb of 1pter-1p36.23 and result in mental retardation, developmental delay, sensorineural hearing loss, seizures, cardiomyopathy and cardiovascular malformations, and distinct facies including large anterior fontanel, deep-set eyes, straight eyebrows, flat nasal bridge, asymmetric ears, and pointed chin. We report five patients with 'atypical' proximal interstitial deletions from 1p36.23-1p36.11 using array-comparative genomic hybridization. Four patients carry large overlapping deletions of approximately 9.38-14.69 Mb in size, and one patient carries a small 2.97 Mb deletion. Interestingly, these patients manifest many clinical characteristics that are different from those seen in 'classical' monosomy 1p36 syndrome. The clinical presentation in our patients included: pre- and post-natal growth deficiency (mostly post-natal), feeding difficulties, seizures, developmental delay, cardiovascular malformations, microcephaly, limb anomalies, and dysmorphic features including frontal and parietal bossing, abnormally shaped and posteriorly rotated ears, hypertelorism, arched eyebrows, and prominent and broad nose. Most children also displayed hirsutism. Based on the analysis of the clinical and molecular data from our patients and those reported in the literature, we suggest that this chromosomal abnormality may constitute yet another deletion syndrome distinct from the classical distal 1p36 deletion syndrome.

Practical synthesis of Shi's diester fructose derivative for catalytic asymmetric epoxidation of alkenes.

[reaction: see text] A practical synthesis of Shi's diester 3 for catalytic asymmetric epoxidations has been developed. The catalyst has been prepared in multigram quantities from D-fructose in four steps with a 66% overall yield. Efficiency, cost, and selectivity aspects of the reagents involved for its preparation have been taken care of during its preparation. The workup procedures have been simplified to the bare minimum, rendering a very practical preparation method. The well-known high efficiency of this catalyst 3 in the epoxidation of alpha,beta-unsaturated carbonyl compounds has also proved to be high in unfunctionalized alkenes.

Detection of high-energy phosphates in cortical bone as an indicator of bone healing and remodelling: use of a rabbit model.

UNLABELLED: PURPOSE; To study high-energy phosphates in cortical bone through experiments on inbred white New Zealand rabbits. METHODS: Tibial fractures were induced in 80 rabbits and then stabilised by screw osteosynthesis. After 3 (group A; n=40) or 7 days (groups B; n=40), the defective tissue was covered by local muscle flaps. At increasing intervals (from 1 to 16 weeks), the screws were removed and the animals were euthanised (n=8 per group). The bone was removed and analysed histomorphologically; adenosine triphosphate (ATP) levels were determined by high-performance liquid chromatography. RESULTS: The mean ATP concentration in healthy cortical bone at 16 weeks was 0.092 (standard error, 0.009) nmol/mg dry mass, which was significantly higher than that in the group with delayed healing: 0.081 (0.011) nmol/mg in group A and 0.005 (0.001) nmol/mg in group B (paired t test, p<0.05). Earlier healing led to lower rates of necrosis (0 vs 38; groups A vs B) and osteomyelitis. CONCLUSION: Early muscle-flap coverage can revascularise the cortical bone, which is reflected in the higher ATP content in the cortical bone measured by high-performance liquid chromatography. Measuring changes of ATP levels can help investigate the metabolism of the pathological bone.

[Locally recurrent rectal carcinoma]. / Lokoregionäres Rezidiv des Rektumkarzinoms.

There is no doubt that surgical treatment with curative intent is warranted in patients with recurrent colorectal cancer. This is also true for locally recurrent rectal cancer. Operative procedures are demanding but may be curative if complete excision can be achieved. Preoperative radiochemotherapy has helped to improve resectability, but operative morbidity is still high. Procedure-related mortality on the other hand is low, at around 3%, and 25% 5-year survival can be expected after complete excision.

Laparoscopic management of hydronephrosis in a woman with rare genitourinary anomalies: unicornous uterus, contralateral agenesis of the kidney, and contralateral ectopic ovary in the groin.

A 40-year-old woman had infected right-sided hydronephrosis and rare genitourinary anomalies-a dextroposed unicornous uterus-as the cause of the hydronephrosis, which had to be mobilized laparoscopically by dissecting the scar tissue. Furthermore, the patient had left-sided agenesis of the kidney and a left-sided ectopic ovary with a rudimentary tube in the inner inguinal canal.