Identification of stable genes in the corpus luteum of lactating Holstein cows in pregnancy and luteolysis: Implications for selection of reverse-transcription quantitative PCR reference genes.

In lactating dairy cattle, the corpus luteum (CL) is a dynamic endocrine tissue vital for pregnancy maintenance, fertility, and cyclicity. Understanding processes underlying luteal physiology is therefore necessary to increase reproductive efficiency in cattle. A common technique for investigating luteal physiology is reverse-transcription quantitative PCR (RT-qPCR), a valuable tool for quantifying gene expression. However, reference-gene-based RT-qPCR quantification methods require utilization of stably expressed genes to accurately assess mRNA expression. Historically, selection of reference genes in cattle has relied on subjective selection of a small pool of reference genes, many of which may have significant expression variation among different tissues or physiologic states. This is particularly concerning in dynamic tissues such as the CL, with its capacity for rapid physiologic changes during luteolysis, and likely in the less characterized period of CL maintenance during pregnancy. Thus, there is a clear need to identify reference genes well suited for the bovine CL over a wide range of physiological states. Whole-transcriptome RNA sequencing stands as an effective method to identify new reference genes by enabling the assessment of the expression profile of the entire pool of mRNA transcripts. We report the identification of 13 novel putative reference genes using RNA sequencing in the bovine CL throughout early pregnancy and luteolysis: RPL4, UQCRFS1, COX4I1, RPS4X, SSR3, CST3, ZNF266, CDC42, CD63, HIF1A, YWHAE, EIF3E, and PPIB. Independent RT-qPCR analyses were conducted confirming expression stability in another set of CL tissues from pregnancy and regression, with analyses performed for 3 groups of samples: (1) all samples, (2) samples from pregnancy alone, and (3) samples throughout the process of CL regression. Seven genes were found to be more stable in all states than 2 traditional reference genes (ACTB and GAPDH): RPS4X, COX4I1, PPIB, SSR3, RPL4, YWHAE, and CDC42. When CL tissues from pregnant animals alone were analyzed, CST3, HIF1A, and CD63 were also identified as more stable than ACTB and GAPDH. Identification of these new reference genes will aid in accurate normalization of RT-qPCR results, contributing to proper interpretation of gene expression relevant to luteal physiology. Furthermore, our analysis sheds light on the effects of luteolysis and pregnancy on the stability of gene expression in the bovine CL.

Comparison of lameness outcomes in horses with acute or chronic digital lameness that underwent magnetic resonance imaging.

Aims: To compare the outcome, in terms of lameness score or return to athletic function, of horses with acute vs. chronic digital lameness that underwent magnetic resonance imaging (MRI) of the distal limb and to compare the proportion of horses that received intra-articular therapy of the distal interphalangeal (DIP) joint and pattern of diagnostic analgesia in these groups. Methods: This is a retrospective study of horses (n = 95) with acute (≤12 weeks; n = 46) or chronic (>12 weeks; n = 49) digital lameness that underwent MRI of the distal limb from 2009-2016, at two equine referral centres in the USA. Criteria for inclusion in the study were that a majority of lameness localised distal to the fetlock, and that lameness assessments for ≥12 months following MRI could be obtained from the medical record or the owner could be interviewed regarding their horse's athletic function. Outcome was characterised by an improvement score where 2 = return to work at a previous or higher level or lameness improved by one grade or more, 1 = return to work at a lower level or lameness improved by less than one grade, and 0 = did not return to work or lameness grade worsened. Whether horses had received intra-articular therapy of the DIP joint and the pattern of diagnostic analgesia prior to MRI was also obtained from medical records or by interviewing the owner. Results: There was a difference (p = 0.004) in the proportion of horses assigned to improvement scores of 0, 1 and 2 between horses with acute or chronic lameness. There was no evidence of a difference in the likelihood of having received intra-articular therapy of the DIP joint prior to MRI between horses with chronic or acute lameness (p = 0.085). Similarly, there was no evidence of a difference in the pattern of diagnostic analgesia prior to MRI between the two groups (p = 0.94). Eighty-two percent of owners of horses with acute and 62% of those with horses with chronic lameness had a positive opinion of the utility of MRI as a diagnostic modality. Conclusion: In a population of horses with digital lameness undergoing MRI, a difference in the outcome, in terms of lameness score or return to athletic function was identified between horses with acute lameness compared to those with chronic lameness. Clinical relevance: Horses with digital lameness that undergo MRI when the lameness is acute may have an improved prognosis due to accurate diagnosis and earlier application of appropriate therapy.

[Do the new molecular assays-microarray and realtime polymerase chain reaction-for dermatophyte detection keep what they promise?] / Halten die neuen molekularen Teste ­ Microarray und Realtime-Polymerasekettenreaktion ­ zum Dermatophytennachweis das, was sie versprechen?

In this study, a novel real-time polymerase chain reaction (PCR) assay (DermaGenius®2.0, PathoNostics BV, Maastricht, The Netherlands) and a recently developed microarray test (EUROArray Dermatomycosis, Euroimmun, Lübeck, Germany) were evaluated regarding their diagnostic specificity to identify dermatophyte DNA. The tests were compared to conventional methods and sequencing. The microarray Dermatomycosis test allows the detection of 50 dermatophytes and definitive identification of 23 dermatophyte species, 6 yeasts and moulds combined in one test. In comparison, real-time PCR is able to identify 11 dermatophytes and one yeast at the species level. Using the EUROArray, 22 out of 24 dermatophyte species were correctly identified. Using real-time PCR, 9 out of the 11 different dermatophytes included in the test kit were correctly identified. Both molecular tests for detection and differentiation of dermatophytes are useful tools for daily clinical practice. The real-time PCR test does not detect as many species, and specificity is slightly lower. However, real-time PCR is a very fast and easy to perform test, especially since no post-PCR step is necessary. Real-time PCR detects the most frequent dermatophytes like T. rubrum, T. interdigitale, and M. canis without any problems. The EUROArray is more elaborate to perform in the lab, due to the hybridization step. However, the EUROArray shows higher specificity and can detect a much broader range of causative agents, including rare species, in dermatomycology.

[Hedgehog fungi in a dermatological office in Munich : Case reports and review]. / Igelpilze in einer Münchner Hautarztpraxis : Fallbeschreibungen und Übersicht.

Patient 1: After contact to a central European hedgehog (Erinaceus europaeus), a 50-year-old female with atopy developed erythrosquamous tinea manus on the thumb and thenar eminence of the right hand. The patient had previously been scalded by hot steam at the affected site. The zoophilic dermatophyte Trichophyton erinacei could be cultured from the hedgehog as well as from scrapings from the woman's skin. Antifungal treatment of the hedgehog was initiated using 2 weekly cycles of itraconazole solution (0.1 ml/kg body weight, BW). In addition, every other day enilconazole solution was used for topical treatment. The patient was treated with ciclopirox olamine cream and oral terbinafine 250 mg daily for 2 weeks, which led to healing of the Tinea manus .Patient 2: An 18-year-old woman presented for emergency consultation with rimmed, papulous, vesicular and erosive crusted skin lesions of the index finger, and an erythematous dry scaling round lesion on the thigh. The patient worked at an animal care facility, specifically caring for hedgehogs. One of the hedgehogs suffered from a substantial loss of spines. Fungal cultures from skin scrapings of both lesions yielded T. erinacei. Treatment with ciclopirox olamine cream and oral terbinafine 250 mg for 14 days was initiated which led to healing of the lesions. Identification of all three T. erinacei isolates from both patients and from the hedgehog was confirmed by sequencing of the internal transcribed spacer (ITS) region of the ribosomal DNA, and of the translation elongation factor (TEF)-1-alpha gene. Using ITS sequencing discrimination between T. erinacei strains from European and from African hedgehogs is possible. T. erinacei should be considered a so-called emerging pathogen. In Germany the zoophilic dermatophyte T. erinacei should be taken into account as causative agent of dermatomycoses in humans after contact to hedgehogs.

[Tinea capitis and onychomycosis due to Trichophyton soudanense : Successful treatment with fluconazole-literature review]. / Tinea capitis und Onychomykose durch Trichophyton soudanense : Erfolgreiche Behandlung mit Fluconazol ­ Literaturübersicht.

Two African girls who moved to Germany only 4 weeks ago presented to the dermatological office with itchy and scaling skin lesions of the scalp and the thighs. The entire scalp of both girls was affected by a white, dry dandruff and a squamous crust. Dry centrifugal spreading erythematosquamous lesions were found on the thighs. The surface of the left thumbnail of the younger girl was whitish. The Blancophor® preparations which were performed under the suspicion of a tinea capitis et corporis and onychomycosis from skin scrapings of the scalp and the thighs, and from the thumbnail of the younger child were positive. Cultivation of three samples from the affected body sites-hair, skin and nail-revealed Trichophyton (T.) soudanense. For confirmation of the species identification, the isolates were subject of sequencing of ITS region of the rDNA and also of the translation elongation factor 1 α (TEF 1 α) gene. The phylogenetic analysis of the strains-the dendrogram of fungal strains-demonstrated the genetic differences between T. soudanense and T. rubrum. In contrast, sequencing of the TEF 1 α gene did not allow any discrimination between T. soudanense and T. rubrum. Both girls were treated orally with fluconazole. For topical treatment of both girls, ciclopirox olamine solution and terbinafine cream were administered, each once daily. After 8 weeks oral fluconazole therapy the dermatomycoses of skin, scalp, and thumbnail of both children were completely healed. Currently, in Germany and Europe, in immigrants from West African countries (e. g., from Angola) dermatophytoses due to T. soudanense have to be expected. Cultural identification of the pathogen is relatively simple. However, only molecular methods allow the exact discrimination of T. violaceum and T. rubrum.

[Characteristic hindlimb lameness presentations in dogs]. / Gangbilder bei Lahmheiten der Hintergliedmasse des Hundes.

INTRODUCTION: Lameness in dogs are caused in about 70% of the cases in the hind limb, whereof about 50% are attributed to the knee joint. The current article shows selected lameness presentations on the basis of typical case studies. The lameness videos can be viewed via the corresponding QR code scanned with your mobile phones and tablets. Supplementary information on the diagnosis and therapy rounds off the cases.

INTRODUCTION: Lors de boiteries chez le chien, la couse est à chercher dans 70% des cas au niveau des postérieurs et pour 50% au niveau du genou. Sur la base de cas typiques choisis, on présente les images de la locomotion qui y sont associées. On peut, en scannant via le QR-Code correspondant sur une téléphone mobile ou une tablette, visualiser les films. Des informations sur la pose de diagnostic et le traitement complètent les cas.

New insights on the antibacterial efficacy of miconazole in vitro.

Miconazole is a broad-spectrum antifungal used in topical preparations. In the present investigation the minimal inhibitory concentration (MIC) of miconazole for eighty wild type strains of gram-positive and gram-negative bacteria isolated from infected skin lesions was assessed using a modified agar dilution test (adapted to CLSI, Clinical Laboratory Standards Institute). 14 ATCC reference strains served as controls. Miconazole was found efficacious against gram-positive aerobic bacteria (n=62 species), the MICs against Staphylococcus (S.) aureus, S. spp., Streptococcus spp. und Enterococcus spp. ranged between 0.78 and 6.25 µg/mL. Interestingly, there were no differences in susceptibility between methicillin-susceptible (MSSA, 3) methicillin-resistant (MRSA, 6) and fusidic acid-resistant (FRSA, 2) S. aureus isolates. Strains of Streptococcus pyogenes (A-streptococci) (8) were found to be slightly more sensitive (0.78-1.563 µg/mL), while for gram-negative bacteria, no efficacy was found within the concentrations tested (MIC >200 µg/mL). In conclusion, for the gram-positive aerobic bacteria the MICs of miconazole were found within a range which is much lower than the concentration of miconazole used in topical preparations (2%). Thus topically applied miconazole might be a therapeutic option in skin infections especially caused by gram-positive bacteria even by those strains which are resistant to antibiotics.

[Tinea barbae profunda due to Trichophyton mentagrophytes after journey to Thailand : Case report and review]. / Tinea barbae profunda durch Trichophyton mentagrophytes nach Thailand-Reise : Fallbeschreibung und Übersicht.

Tinea barbae represents a very rare dermatophytosis. We report on a tinea barbae profunda following a journey to Southeast Asia. After travel to Thailand, a businessman was affected by a foudroyant proceeding abscessing infection of the upper lip and beard area. The initial therapy with oral acyclovir and oral ciprofloxacin, which later was changed to ampicillin plus sulbactam, intravenously, was unsuccessful. In a biopsy sample, histologically, with Grocott-Gomori's methenamine silver stain, fungal mycelium was apparent in the tissue. Thereupon, terbinafine 250 mg was given for 4 weeks, topically, a 1% ciclopiroxolamine-containing cream. In fungal culture, T. mentagrophytes were found to grow. Meanwhile, the patient's German wife suffered from a tinea faciei. From skin scrapings from the cheek, T. mentagrophytes was also cultivated. This zoophilic dermatophyte was identical with other zoophilic strains of T. mentagrophytes currently found in Germany, which were also acquired in Thailand. The patient had contact with Thai female sex workers who must be considered as a source of infection of the dermatophytosis. There was no animal contact, neither in Thailand, nor in Germany. The infection chain of the dermatophytosis from Thailand probably reached from a female sex worker via the here described patient to his wife in Germany. This pathway of infection has been known for 1 or 2 years, but until now, in Germany, Switzerland, and Austria exclusively via pubogenital infections (tinea genitalis profunda) due to T. mentagrophytes after journeys to Southeast Asia. For treatment, oral antifungal agents should be used, first of all terbinafine, alternatively fluconazole or itraconazole.

[Simple repair of Monteggia fractures in the cat]. / Einfache Methode zur Stabilisierung von Monteggia Frakturen bei der Katze.

INTRODUCTION: Monteggia fractures are rare traumatic injuries in cats and dogs. Ulna repair is mostly achieved by plates. Stable radial head fixation however is challenging. Temporary position screws have to be removed due to rotation forces in the elbow joint causing screw loosening. We present a novel and simple method using a sling technique, which holds the radial head in its physiological position and allows normal elbow movement.

Aggregation and a strong Allee effect in a cooperative outbreak insect.

Most species that are negatively impacted when their densities are low aggregate to minimize this effect. Aggregation has the potential to change how Allee effects are expressed at the population level. We studied the interplay between aggregation and Allee effects in the mountain pine beetle (Dendroctonus ponderosae Hopkins), an irruptive bark beetle that aggregates to overcome tree defenses. By cooperating to surpass a critical number of attacks per tree, the mountain pine beetle is able to breach host defenses, oviposit, and reproduce. Mountain pine beetles and Hymenopteran parasitoids share some biological features, the most notable of which is obligatory host death as a consequence of parasitoid attack and development. We developed spatiotemporal models of mountain pine beetle dynamics that were based on the Nicholson-Bailey framework but which featured beetle aggregation and a tree-level attack threshold. By fitting our models to data from a local mountain pine beetle outbreak, we demonstrate that due to aggregation, attack thresholds at the tree level can be overcome by a surprisingly low ratio of beetles per susceptible tree at the stand level. This results confirms the importance of considering aggregation in models of organisms that are subject to strong Allee effects.

Calculation of the static and dynamical correlation energy of pseudo-one-dimensional beryllium systems via a many-body expansion.

Due to the importance of both static and dynamical correlation in the bond formation, low-dimensional beryllium systems constitute interesting case studies to test correlation methods. Aiming to describe the whole dissociation curve of extended Be systems we chose to apply the method of increments (MoI) in its multireference (MR) formalism. To gain insight into the main characteristics of the wave function, we started by focusing on the description of small Be chains using standard quantum chemical methods. In a next step we applied the MoI to larger beryllium systems, starting from the Be6 ring. The complete active space formalism was employed and the results were used as reference for local MR calculations of the whole dissociation curve. Although this is a well-established approach for systems with limited multireference character, its application regarding the description of whole dissociation curves requires further testing. Subsequent to the discussion of the role of the basis set, the method was finally applied to larger rings and extrapolated to an infinite chain.

Vemurafenib-induced hyperkeratosis of the areola treated with topical adapelene.

We present a rare condition, hyperkeratosis of the areola, induced by vemurafenib. Only a few papers have described an association of BRAF inhibitors with hyperkeratosis of the areola and/or nipple. Vemurafenib is a selective BRAF inhibitor used in patients with unresectable or metastatic melanoma who are positive for the V600 mutation. This drug has been associated with numerous cutaneous side effects, both benign and malignant. We report a male patient with vemurafenib-induced hyperkeratosis of the areola managed successfully with a topical retinoid, and describe for the first time a treatment for this side effect.

Analysis of traction-free assumption in high-resolution EBSD measurements.

The effects of using a traction-free (plane-stress) assumption to obtain the full distortion tensor from high-resolution EBSD measurements are analyzed. Equations are derived which bound the traction-free error arising from angular misorientation of the sample surface; the error in recovered distortion is shown to be quadratic with respect to that misorientation, and the maximum 'safe' angular misorientation is shown to be 2.7 degrees. The effects of localized stress fields on the traction-free assumption are then examined by a numerical case study, which uses the Boussinesq formalism to model stress fields near a free surface. Except in cases where localized stress field sources occur very close to sample points, the traction-free assumption appears to be admirably robust.

Impact of intraoperative microperfusion assessment with Pinpoint Perfusion Imaging on surgical management of laparoscopic low rectal and anorectal anastomoses.

AIM: Inadequate intestinal blood flow may contribute to anastomotic leakage accounting for substantial morbidity and mortality in colorectal surgery. Precise intraoperative assessment of microperfusion may have an impact on the surgeons intraoperative management and leakage rate. METHOD: In this single center observational study we implemented and integrated intraoperative indocyanin green (ICG) based microperfusion assessment of anastomosis with Pinpoint Perfusion Imaging in a series of consecutive rectal cancer patients who underwent laparoscopic anterior and lower anterior resection with primary anastomosis during a 5-months period. RESULTS: We could demonstrate the feasibility and safety of intraoperative fluorescence angiography for colorectal microperfusion assessment. Technology implementation was immediately successful. No adverse effects have been documented related to fluorescent dye. Microperfusion angiography of the colon succeeded in all cases and assessment of perfusion imaging influenced surgical decision making in 28% of the patients, of which all patients showed primary healing of the anastomosis. We found a leakage rate of 6% with one leakage of a coloanal anastomosis in all patients. CONCLUSION: Fluorescence angiography is an accurate tool for assessing microperfusion and is most likely associated with improved outcomes with regard to anastomotic healing.

The diverse application of laser hair removal therapy: a tertiary laser unit's experience with less common indications and a literature overview.

We describe the diversity of indications for laser hair removal (LHR) therapy and compare our experience with the literature. Patients' case notes referred to the Birmingham Regional Skin Laser Centre between 2003 and 2011 for laser hair removal, with indications other than hirsutism, were reviewed retrospectively. Thirty-one treated patients with the following indications were identified: hair-bearing skin grafts/flaps, intra-oral hair-bearing flap, Becker's naevus, localised nevoid hypertrichosis, peristomal hair-bearing skin, scrotal skin prior to vaginoplasty in male-to-female (MTF) gender reassignment, pilonidal sinus disease (PSD), pseudofolliculitis barbae (PFB) and hidradenitis suppurativa (HS). Seven patients with the following indications have been reported before: intra-oral hair-bearing graft, naevoid hypertrichosis and peristomal hair-bearing skin. A clinical review of the evidence available for each indication is provided. Our experience and that in the published literature suggest that LHR is a safe, well-tolerated and effective treatment modality for the indications we report, leading to significant symptom and functional improvement with high patient satisfaction. LHR appears effective in the treatment of chronic inflammatory conditions such as PSD, PFB and HS, particularly at an early disease stage. We aim to increase awareness of the diversity of laser hair removal indications and add evidence to the medical literature of the wide range of indications for this useful treatment modality.

[Measures to improve the health situation of patients with rare diseases in Germany. A comparison with the National Action Plan]. / Maßnahmen zur Verbesserung der gesundheitlichen Situation von Menschen mit seltenen Erkrankungen in Deutschland. Ein Vergleich mit dem Nationalen Aktionsplan.

BACKGROUND: Approximately 4 million patients with a rare disease live in Germany. The medical care of these patients is problematic because of the rarity and heterogeneity of different clinical pictures. The Federal Ministry of Health has therefore published a research report on "Measures to improve the health situation of people with rare diseases in Germany" in 2009. OBJECTIVE: The aim of this paper is to present the main recommendations of this research report and relate it to current developments in the field of medical care for people with rare diseases. METHODOLOGY: The care situation of patients with rare diseases was determined using questionnaires, expert interviews and focus group discussions with representatives of patients, service providers and stakeholders from the health institutions. RESULTS: The main range of actions that have been identified in the research report were centre and network formation, specialized forms of medical care, diagnosis and treatment, information and experience exchange, performance fees and reimbursement of the costs, guidelines and patient pathways, the research, the implementation of a National Action Alliance and the development of a National Action Plan. DISCUSSION: In March 2010 a National Action League for People with Rare Diseases (NAMSE) was founded. The NAMSE created a national plan of action for people with rare diseases for improving medical care in the field of rare diseases which was approved by the Federal Government in August 2013. Thus, two important areas of the research report have already been implemented. In a comparison of the areas of activity of the research report with those of the National Action Plan it becomes clear that priorities will be in the context of health services research in rare diseases, for example the introduction of centres of reference for rare diseases, measures to accelerate the diagnostic process and the promotion of research and information management in the future.

Comparative investigations on the upper respiratory tract in Norwich terriers, brachycephalic and mesaticephalic dogs.

For some time Norwich terriers have been known to suffer from respiratory problems. In order to assign this weakness to a pathophysiology, 23 terriers were examined clinically, with laryngoscope and with rhinomanometry. In addition their skulls were dimensioned on radiographs. Widened nostrils, overlong soft palates and the everted laryngeal pouches were consistent with brachycephalic syndrome. Resistance values in the nasal passage corresponded to the appearance in brachycephalic dogs. Skull measurements gave inconsistent results, because length to width ratios and craniofacial angles denoted mesaticephaly, whereas the facial to cranial length ratios (S-index = 0.65) lay in the brachycephalic sector. It can be concluded, that the Norwich terrier breed is in transition towards brachycephaly with some individual dogs already suffering from the brachycephalic syndrome. Breeders are requested to introduce necessary counter measures.

On sait depuis longtemps déjà que le Norwich Terrier souffre de plus en plus de problèmes respiratoires. Afin de pouvoir donner une explication pathophysiologique à ces problèmes, on a examiné 23 Norvich Terriers sur le plan clinique, laryngoscopique, rhino-manométrique ainsi qu'avec des radiographies du crâne. Des narines larges, un voile du palais trop long et des ventricules laryngés éversés, de même que des mesures de résistance augmentées dans les conduits nasaux correspondaient à l'image du syndrome brachycéphale. Les mesures du crâne présentaient un tableau non uniforme. L'index longueur-largeur et l'angle cranio-facial correspondaient à ceux de chiens mésocéphales, alors que l'index splanchnocrâne-neurocrâne (Index S de 0.65) était clairement dans la zone du brachicéphalisme. On peut en déduire que le Norwich Terrier se trouve sur le chemin du brachicéphalisme et que certains chiens souffrent déjà d'un syndrome brachycéphale. Les éleveurs doivent donc prendre des mesures pour lutter contre cette évolution.