Tissue engineering at the dentin-pulp interface using human treated dentin scaffolds conditioned with DMP1 or BMP2 plasmid DNA-carrying calcium phosphate nanoparticles.

Hard dental tissue pathologies, such as caries, are conventionally managed through replacement by tooth-colored inert biomaterials. Tissue engineering provides novel treatment approaches to regenerate lost dental tissues based on bioactive materials and/or signaling molecules. While regeneration in the form of reparative dentin (osteo-dentin) is feasible, the recapitulation of the tubular microstructure of ortho-dentin and its special features is sidelined. This study characterized in vitro, and in vivo human EDTA-treated, freeze-dried dentin matrices (HTFD scaffolds) conditioned with calcium phosphate nanoparticles (NPs) bearing plasmids encoding dentinogenesis-inducing factors (pBMP2/NPs or pDMP1/NPs). The uptake and transfection efficiency of the synthesized NPs on dental pulp stem cells (DPSCs) increased in a concentration- and time-dependent manner, as evaluated qualitatively by confocal laser microscopy and transmission electron microscopy, and quantitatively by flow cytometry, while, in parallel, cell viability decreased. HTFD scaffolds conditioned with the optimal transfectability-to-viability concentration at 4 µg Ca/mL of each of the pBMP2/NPs or pDMP1/NPs preserved high levels of cell viability, evidenced by live/dead staining in vitro and caused no adverse reactions after implantation on C57BL6 mice in vivo. HTFD/NPs constructs induced rapid and pronounced odontogenic shift of the DPSCs, as evidenced by relevant gene expression patterns of RunX2, ALP, BGLAP, BMP-2, DMP-1, DSPP by real-time PCR, and acquirement of polarized meta-mitotic phenotype with cellular protrusions entering the dentinal tubules as visualized by scanning electron microscopy. Taken together, HTFD/NPs constitute a promising tool for customized reconstruction of the ortho-dentin/odontoblastic layer barrier and preservation of pulp vitality. STATEMENT OF SIGNIFICANCE: In clinical dentistry, the most common therapeutic approach for the reconstruction of hard dental tissue defects is the replacement by resin-based restorative materials. Even modern bioactive materials focus on reparative dentinogenesis, leading to amorphous dentin-bridge formation in proximity to the pulp. Therefore, the natural microarchitecture of tubular ortho-dentin is not recapitulated, and the sensory and defensive role of odontoblasts is sidelined. This study approaches the reconstruction at the dentin-pulp interface using a construct of human treated dentin (HTFD) scaffold and plasmid-carrying nanoparticles (NPs) encoding dentinogenic factors (DMP-1 or BMP-2) with excellent in vitro and in vivo properties. As a future perspective, the HTFD/NPs constructs could act as bio-fillings for personalized reconstruction of the dentin-pulp interface.

Unbreakable? An analysis of the fragility of randomized trials that support diabetes treatment guidelines.

AIMS: To analyze the robustness of randomized controlled trials (RCTs) referenced in the American Diabetes Association's Standards of Medical Care in Diabetes-2017 using the Fragility Index (FI) and Fragility Quotient (FQ). METHODS: We performed a systematic survey of all RCTs referenced in the Standards of Medical Care in Diabetes-2017. One investigator screened for trials and then recorded data from them, including sample size per group, event rate per group, and the dichotomous outcome analyzed by trialists. The FI and FQ for each outcome were calculated. Outcomes were then surveyed to determine whether the outcome used for analysis aligned with the guideline recommendation. RESULTS: Thirty-five RCTs were included in this study. Thirty-three of 35 (94%) FIs were based on the trial outcomes referenced in the clinical practice guideline. The median sample size was 2548 participants ([IQR], 522-6946). The median total number of events for each outcome was 403 (IQR, 86-969). Nineteen (54%) P Values were below 0.05, 8 (22.4%) were below 0.01. The median FI for all trials was 16 (IQR, 4-29). The median FQ was 0.007 (IQR, 0.003-0.014). FI was not related to risk of bias or Science Citation Index but was significantly correlated with sample size (for significant trials; r=0.77, P < .001, for neutral trials; r=0.76, P < .001). CONCLUSIONS: We found that the robustness of RCTs varied, but on the whole were not robust in nature. Most trials demonstrated a modest FI and FQ. As a result, conclusions drawn from these trials should take this information into account.

Augmentation of Transient Donor Cell Chimerism and Alloantigen-Specific Regulation of Lung Transplants in Miniature Swine.

Donor alloantigen infusion induces T cell regulation and transplant tolerance in small animals. Here, we study donor splenocyte infusion in a large animal model of pulmonary transplantation. Major histocompatibility complex-mismatched single lung transplantation was performed in 28 minipigs followed by a 28-day course of methylprednisolone and tacrolimus. Some animals received a perioperative donor or third party splenocyte infusion, with or without low-dose irradiation (IRR) before surgery. Graft survival was significantly prolonged in animals receiving both donor splenocytes and IRR compared with controls with either donor splenocytes or IRR only. In animals with donor splenocytes and IRR, increased donor cell chimerism and CD4(+) CD25(high+) T cell frequencies were detected in peripheral blood associated with decreased interferon-γ production of leukocytes. Secondary third-party kidney transplants more than 2 years after pulmonary transplantation were acutely rejected despite maintained tolerance of the lung allografts. As a cellular control, additional animals received third-party splenocytes or donor splenocyte protein extracts. While animals treated with third-party splenocytes showed significant graft survival prolongation, the subcellular antigen infusion showed no such effect. In conclusion, minipigs conditioned with preoperative IRR and donor, or third-party, splenocyte infusions may develop long-term donor-specific pulmonary allograft survival in the presence of high levels of circulating regulatory T cells.

Comparison of auricular and rectal temperature measurement in normothermic, hypothermic, and hyperthermic dogs.

OBJECTIVE: Measurement of rectal temperature is the most common method and considered gold standard for obtaining body temperature in dogs. So far, no study has been performed comparing agreement between rectal and auricular measurements in a large case series. The purpose of the study was to assess agreement between rectal and auricular temperature measurement in normothermic, hypothermic, and hyperthermic dogs with consideration of different environmental conditions and ear conformations. MATERIALS AND METHODS: Reference values for both methods were established using 62 healthy dogs. Three hundred dogs with various diseases (220 normothermic, 32 hypothermic, 48 hyperthermic) were enrolled in this prospective study. Rectal temperature was compared to auricular temperature and differences in agreement with regard to environmental temperature, relative humidity, and different ear conformations (pendulous versus prick ears) were evaluated using Pearson's correlation coefficient and Bland-Altman analysis. RESULTS: Correlation between rec- tal and auricular temperature was significant (r: 0.892; p  <  0.01). However, Bland-Altman plots showed an inacceptable variation of values (bias: 0.300 °C; limits of agreement: -0.606 to 1.206 °C). This variation was above a maximal clinical tolerance of 0.3 °C, which was established by experts' opinion (n = 16). Relative humidity had a significant influence (p   =   0.001), whereas environmental temperature did not. CONCLUSION: Variation between the two methods of measuring body temperature was clinically unacceptable. CLINICAL RELEVANCE: Although measurement of auricular temperature is fast, simple, and well tolerated, this method provides a clinically unacceptable difference to the rectal measurement.

[Disseminated intravascular coagulopathy in a dog with Angiostrongylus vasorum infection]. / Disseminierte intravasale Koagulopathie bei einem Hund mit Angiostrongylus-vasorum-Infektion.

A 2-year-old female spayed Epagneul-Breton dog was presented with ecchymoses, but an undisturbed general condition. Clinical examination additionally revealed petechia and a haematoma. Travel history included Italy and Denmark. Laboratory abnormalities were moderate thrombocytopenia, prolonged PT, aPTT and TT, and elevated d-dimers. Initial therapy consisted of plasma transfusions, fluids, doxycycline and famotidine administration. Babesiosis, ehrlichiosis, leishmaniosis, dirofilariosis or anaplasmosis could not be confirmed. Abdominal ultrasound was unremarkable, while thoracic radiographs showed a bronchointerstitial pattern. Faecal samples collected over 3 days were positive for Angiostrongylus vasorum after examination using the Baermann lungworm test. The A. vasorum infection was successfully treated with fenbendazole, whereupon thrombocytopenia and prolonged coagulation times were resolved. In regions of low prevalence, an infection with A. vasorum should also be considered as a differential diagnosis in dogs with coagulation abnormalities. Respiratory signs can be absent with this disease. The patient may have acquired the infection abroad or in Germany.

[Feline panleukopenia - different course of disease in cats younger than versus older than 6 months of age?]. / Feline Panleukopenie - differierender Krankheitsverlauf bei Katzen im Alter von unter bzw. über 6 Monaten?

OBJECTIVE: The aim of this study was to evaluate differences between young cats (< 6 months) and adult cats (≥ 6 months) with feline panleukopenia with respect to clinical signs, laboratory abnormalities, environmental conditions, vaccination status, and outcome. MATERIAL AND METHODS: Medical records of 244 cats diagnosed with panleukopenia between 1990 and 2007 at the Clinic of Small Animal Medicine, Ludwig-Maximilians University of Munich, Germany, were evaluated retrospectively. Cats that tested positive for feline panleukopenia virus (FPV) via electron microscopy, polymerase chain reaction (blood, faeces), antigen ELISA (faeces), or that had histopathological lesions consistent with panleukopenia at necropsy were included. Cats were excluded if they had been vaccinated against FPV within 3 weeks before admission. RESULTS: In total 43.3% of cats were older than 6 months. There was no statistically significant difference between the two age groups regarding outcome, breed, sex, environmental conditions, vaccination status, clinical signs, and laboratory parameters with the exception of haematocrit: cats < 6 months had significantly lower haematocrit on the day of presentation than cats ≥ 6 months. CONCLUSIONS AND CLINICAL RELEVANCE: Feline panleukopenia is predominantly in young cats, but older cats can also suffer from the disease. Although young cats are at a higher risk of infection, cats at the age of < 6 months suffering from clinical disease do not have a higher risk of death. Clinical presentation, laboratory abnormalities, prognosis, and outcome did not differ significantly between cats younger versus older than 6 months of age.

Treatment of aseptic dogs with hemorrhagic gastroenteritis with amoxicillin/clavulanic acid: a prospective blinded study.

BACKGROUND: Antibiotics generally are recommended to treat hemorrhagic gastroenteritis (HGE). Inappropriate use of antibiotics may promote risk of antimicrobial resistance and unnecessary adverse drug reactions. The necessity of antimicrobial therapy in dogs with HGE has not been demonstrated. OBJECTIVE: The purpose of this prospective, placebo-controlled, blinded study was to evaluate whether treatment with amoxicillin/clavulanic acid improves the clinical course and outcome of HGE in dogs that show no signs of sepsis. ANIMALS: The study included 60 dogs diagnosed with HGE between 2007 and 2009 at the Clinic of Small Animal Medicine, LMU University of Munich, Germany. The inclusion criterion was the presence of acute hemorrhagic diarrhea (<3 days). Dogs pretreated with antibiotics, with signs of sepsis, or diagnosed with any disease known to cause bloody diarrhea were excluded from the study. METHODS: Patients were randomly divided into treatment (amoxicillin/clavulanic acid for 7 days) and placebo groups. To evaluate treatment efficacy, severity of clinical signs (based on a newly developed HGE index), duration of hospitalization, and mortality rate were compared between the 2 groups. RESULTS: Fifty-three of 60 dogs completed the study. No significant difference between treatment groups concerning mortality rate, dropout rate, duration of hospitalization, or severity of clinical signs, either on any individual day or over the course of disease, was observed. CONCLUSIONS AND CLINICAL IMPORTANCE: In some dogs with HGE that show no signs of sepsis, antibiotics may not change the case outcome or time to recovery.

Prognostic factors in cats with feline panleukopenia.

BACKGROUND: Feline panleukopenia is a highly contagious and often lethal disease. OBJECTIVE: The purpose of the study was to identify prognostic factors for survival of cats with panleukopenia. ANIMALS: Between 1990 and 2007, 244 cats were diagnosed with panleukopenia in the Clinic of Small Animal Medicine, LMU University of Munich, Germany. Diagnosis was established by electron microscopy, polymerase chain reaction of feces or blood, antigen ELISA of feces, pathognomonic histopathological lesions at necropsy, or some combination of these procedures. METHODS: Medical records of each cat were evaluated retrospectively. RESULTS: Survival rate was 51.1%. No significant correlation was found between outcome and living conditions, age, vaccination status (unvaccinated versus one or more vaccines administered), or severity of clinical signs. However, of the vaccinated cats, none had received a vaccine later than 12 weeks of age as a kitten. Nonsurvivors had significantly lower leukocyte and thrombocyte counts at presentation compared with survivors. The relative risk of death for patients with <1,000/µL leukocytes was 1.77 times as high as in patients with a leukocyte count of 1,000-2,500/µL (P=.038), and 1.85 times as high as in patients with >2,500/µL leukocytes (P=.001). The likelihood of a fatal outcome was higher when serum albumin concentration was <30 g/L or serum potassium concentration <4 mmol/L. CONCLUSIONS AND CLINICAL IMPORTANCE: Vaccination strategies that do not include vaccination of kittens beyond 12 weeks of age may not be adequate to prevent panleukopenia. Leukopenia, thrombocytopenia, hypoalbuminemia, and hypokalemia are negative prognostic factors in cats with panleukopenia.

[PDCA cyclus and morbidity and mortality conference as a basic tool for reduction of wound infection in colorectal surgery]. / PDCA-Zyklus und Morbiditäts- und Mortalitätskonferenz als Basis der Wundinfektionsreduktion in der Kolorektalchirurgie.

BACKGROUND: Morbidity and mortality conferences (M + M) are used for the discussion of errors and thereby further education in surgery. However, it is not clear whether this kind of quality assurance has any influence on the results of surgery. MATERIAL AND METHODS: We investigated whether M + M as part of PDCA-cylce (plan, do, check, act) with prospectively collected quality data, fixing future goals using benchmarking data and defining a strategy to reach these goals can lead to an increase in quality. RESULTS: Using surveillance data n = 673 colorectal resections were analysed. Between 2003 and 2008 we found a wound infection rate of 5.6 %. Comparing 2003-2005 (period of strategy development using M + M) and 2006-2008 (period of strategy conversion) the frequency decreased from 7.8 % to 3.5 % (p = 0.012). Thereby the risk factors for wound infection (ASA, wound classification, duration of surgery and wound infection score) were not different. CONCLUSIONS: M + M as part of this PDCA cycle resulted in a clear increase in surgical quality.

Enhanced expression and clinical significance of CC-chemokine MIP-3 alpha in hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most frequent visceral neoplasms worldwide. Using RT-PCR, ELISA, microdissection and immunohistochemistry, we investigated the expression profiles of CCL19, CCL20, CCL21 and CXCL12 and their receptors in tumourous and tumour neighbouring tissues from patients with HCC and in nonmalignant liver lesions, respectively. All chemokines were found to be expressed in normal liver and HCC tissues, yet CCL20 was the only chemokine showing significant upregulation in HCC tissues. Clinicopathological analysis revealed a distinct increase in CCL20 expression rates in HCC tissues of grade III tumours in comparison to HCC tissues from grade II tumours. On mRNA level, only chemokine receptor CCR6 revealed significant upregulation in HCC tissues. However, immunohistochemical studies indicated a marked CCR6 expression accumulated in a streak of normal cells along the tumour invasion front in all our HCC specimens which could provide a stimulative signal for the tumour to further expand. The present findings show significant overexpression of CCL20 in the tumour tissues and marked overexpression of the corresponding receptor CCR6 in the tumour invasion front of HCC patients in comparison to normal liver. Moreover, CCL20 expression was found to correlate with tumour grade and therefore, we suggest that the CCL20/CCR6 system may be involved in hepatocarcinogenesis.

[L-2-hydroxyglutaric aciduria -- a rare cause of macrocephaly]. / L-2-Hydroxy-Glutarazidurie -- eine seltene Differenzialdiagnose der Makrozephalie.

We report on a 9-year-old girl who was referred to our department because of increasing macrocephaly and school problems. The neurological examination disclosed mild cerebellar dysfunction and positive pyramidal tract signs. An MRI of the brain revealed extensive signal alterations of the white matter. Biochemical investigations established the diagnosis of L-2-hydroxyglutaric aciduria which has to be kept in mind as a rare cause of macrocephaly.

Congenital adrenal hyperplasia - how to improve the transition from adolescence to adult life.

Congenital adrenal hyperplasia (CAH) is caused by a defect in the biosynthesis of cortisol that results in maximal activity of the hypothalamic-pituitary adrenal axis with hyperplasia of the adrenals and hyperandrogenism due to the accumulation of androgen precursors. In the salt-wasting subtype of the disorder, which accounts for appr. 75 % of patients with classical CAH, patients are unable to synthesise sufficient amounts of aldosterone and are prone to life-threatening salt-losing crises, whereas the simple virilising form is predominantly characterized by clitoris hypertrophy and posterior labial fusion. In addition, a non-classical variant can be discerned which in most cases is diagnosed at the time of puberty or early adolescence when hirsutism and menstrual irregularities may occur. The vast majority of CAH patients have 21-hydroxylase deficiency (90 - 95 %). Less common forms, such as 11beta-hydroxylase deficiency, will not be discussed in this review. Unfortunately, a considerable number of CAH patients is lost to regular and competent follow-up once they move out of paediatric care. This is most probably the result of insufficient co-operation between paediatric and adult endocrinologists at the time of transition from adolescence to adulthood. Furthermore, there is a lack of clinical guidance regarding psychosexual development in these patients. In this overview we will focus on special aspects of CAH treatment in adolescence and adulthood, and report on our 10-year experience with a transfer system for endocrine patients from paediatric to internal medical care, known as the "Kieler Modell". For practical purposes, we here provide charts for follow-up of CAH patients that can be adapted for use in any endocrine outpatient clinic.

Epilepsy surgery outcome in children with focal epilepsy due to tuberous sclerosis complex.

Tuberous sclerosis complex (TSC) is frequently associated with focal epilepsy due to cerebral tubers. Seizures are the first symptoms in most patients with brain involvement. These epilepsies are frequently severe, drug-resistant and may have a negative impact on the child's global development. Although most epilepsies are multicentric, these patients may be candidates for epilepsy surgery, if it is possible to determine a leading epileptogenic tuber. Nine patients with TSC were examined with long-term video-EEG monitoring, different neuroimaging techniques and neuropsychological tests. A main epileptogenic tuber could be identified in all of our patients. We found good correlations between neuroimaging and EEG. Surgery was performed in eight patients. Seizure outcome was good in all. Two patients became seizure-free, one patient had a single prolonged seizure five days postoperatively, four patients had a significant seizure reduction of more than 75 % and one patient had a seizure reduction of more than 50 % after surgery without additional neurologic deficits. In conclusion, patients with TSC and drug-resistant epilepsy may benefit from epilepsy surgery with reduction in frequency and severity of seizures as well as improved mental and behavioural development leading to a better quality of life. In view of recent developments in functional and metabolic imaging, primary epileptogenic lesions will be more easily detectable in patients with diffuse brain involvement in TSC and surgical treatment may be more specifically applied at an earlier age to a selected subgroup of patients with this disorder.

Chronic effects of ACE-inhibition (quinapril) and angiotensin-II-type-1 receptor blockade (losartan) on atrial natriuretic peptide in brain nuclei of rats with experimental myocardial infarction.

Alterations of the central nervous system may be important for imbalance of cardiovascular and fluid regulation in heart failure. The central renin-angiotensin and atrial natriuretic peptide (ANP) systems act as mutual antagonists. The effects of angiotensin converting enzyme (ACE) inhibition (quinapril, 6 mg/kg/day) and angiotensin II type 1 (AT1) receptor blockade (losartan, 10 mg/kg/day) on ANP levels in 18 selected, microdissected brain nuclei were determined in sham-operated rats and rats with left ventricular dysfunction 8 weeks after myocardial infarction (MI). Plasma ANP tended to increase in MI rats and was further increased by quinapril. ANP was decreased in 12 brain areas of MI rats. ANP concentration was also significantly decreased by quinapril in six brain nuclei including subfornical organ and organum vasculosum laminae terminalis (areas lacking blood-brain barrier), and by losartan in 16 brain nuclei outside and within the blood-brain barrier in sham operated rats. However, both quinapril and losartan prevented a further reduction of central ANP as a result of myocardial infarction. These data suggest that there are effects on central ANP that result from chronic left ventricular dysfunction as well as an ACE-inhibitor and AT1-antagonist. Mechanisms and consequences of central ANP depression remain unclear. They could, however, support systemic vasoconstriction and sodium and fluid retention.

Light scattering and ink penetration effects on tone reproduction.

Light scattering, or the so-called Yule-Nielsen effect, and ink penetration into the substrate paper play important roles in tone reproduction. We develop a framework in which the influences of both of these effects on the reflectance and tristimulus values of a halftone sample are investigated. The properties of the paper and the ink and their bilateral interaction can be parameterized by the reflectance Rp(o) of the substrate paper, the transmittance Ti of the ink layer, the parameter gamma describing the ink penetration, and p describing the Yule-Nielsen effect. We derive explicit expressions that relate the reflectance of the ink dots (Ri), the paper (Rp) and the halftone image (R) as functions of these parameters. We also describe the optical dot gain as a function of these parameters. We further demonstrate that ink penetration leads to a decrease in optical dot gain and that scattering in the paper results in the printed image's being viewed as more saturated in color.

Rehabilitation nursing in home care.

A randomized sample of 83 members of the Association of Rehabilitation Nurses' (ARN's) Home Health Special Interest Group (SIG) responded to a survey in 1998 and 1999 to determine the role intensity of rehabilitation nurses in home care. An instrument was developed that was based on role descriptions formulated by ARN. Significant differences in the roles were reported for caregiver, case manager, counselor, family-client educator, advocate, administrator, student/staff educator, and researcher. Rewards of home nursing included one-to-one interaction with clients, teaching opportunities, promotion of function, nurse autonomy, and seeing rehabilitation results. Difficulties included poor interdisciplinary coordination, budget restrictions, lack of understanding of rehabilitation nursing, and inadequate home aides. Differences between inpatient and home rehabilitation nursing included less equipment and resources and increased levels of responsibility in the home. Barriers for the transition to home rehabilitation nursing included interdisciplinary team communication, reimbursement standards and documentation, time management, autonomous nursing roles, and separation from help or emergency services.

Management of side effects and complications in medical abortion.

Side effects are an expected part of medical abortion; some, such as pain and bleeding, result from the abortion process itself and are generally managed with orally administered analgesics and counseling. True medication side effects most commonly include nausea, vomiting, diarrhea, and warmth or chills. Complications of medical abortion usually represent an extreme or severe side effect. Large series have reported transfusion rates of <1%. Because of the infrequency of uterine instrumentation, postabortal endometritis appears to be rare with medical abortion. As with early surgical abortion, the clinician must remain aware of the possibility for ectopic pregnancy. Overall approximately 2% to 10% of patients will require surgical intervention for control of bleeding, resolution of incomplete expulsion, or termination of a continuing pregnancy. Understanding the types of side effects and complications that can occur will enable the clinician to counsel patients properly as well as to understand when medical intervention is necessary during the medical abortion process.

Advanced practice clinicians and medical abortion: increasing access to care.

Early abortion is one of the safest and most common surgical procedures in the United States, yet it carries a significant stigma for both women and health care providers. Due to a number of factors, including political maneuvering and antichoice violence, the availability of physicians trained and willing to provide abortion has decreased dramatically over the past decades, thereby compounding the obstacles to early termination of pregnancy. At the same time, roles for nurse-midwives, nurse practitioners, and physician assistants have expanded in primary care as well as in specialty practice. Medical abortion is generating renewed interest in abortion care and is potentially more accessible to providers of primary care, women's health, and family planning services. Its provision by qualified advanced practice clinicians working with their physician colleagues promises to significantly improve the delivery of abortion services to women.