[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.

An interactive feedback system for increasing hand antisepsis adherence in stationary intensive care.

BACKGROUND: Pathogens causing infections are in many cases transmitted via the hands of personnel. Thus, hand antisepsis has strong epidemiological evidence of infection prevention. Depending on various factors, hand antisepsis adherence ranges between 9.1% and 85.2%. AIM: To evaluate a new transponder system that reminded medical staff to use an alcohol-based hand rub based on indication by giving real-time feedback, to detect hand antisepsis adherence. METHODS: The monitoring system consisted of three components: a portable transponder detecting alcohol-based hand rub and able to give feedback; a beacon recognizing entries to and exits from the patient's surroundings; and a sensor placed at the hand-rub dispensers to count the number of hand rubs. With these components, the system provided feedback when hand antisepsis was not conducted although it was necessary according to moments 1, 4, and 5 of hand antisepsis. Adherence was measured in two use-cases with five phases, starting with the baseline measurement followed by intervention periods and phases without intervention to test the sustainability of the feedback. FINDINGS: Using the monitoring system, hand antisepsis adherence was increased by up to 104.5% in comparison to the baseline measurement. When the intervention ceased, however, hand antisepsis adherence decreased to less than or equal to the baseline measurement. CONCLUSION: A short-term intervention alone is not sufficient to lead to a long-term change in hand antisepsis adherence. Rather, permanent feedback and/or the integration in a multi-modal intervention strategy are necessary.

Safety and Efficacy of Induction Therapy With Thymoglobulin in AB0-Incompatible Kidney Transplantation.

INTRODUCTION: Data suggest an additional role of T cells in antibody-mediated rejections. In 2001 a protocol for AB0-incompatible kidney transplantation based on B-cell-depleting anti-CD20 antibody rituximab, antigen-specific blood group IgG immunoadsorption, intravenous immunoglobulins, and triple immunosuppression was introduced in Europe, which used induction therapy with the use of interleukin-2 receptor antibody (IL2-RA) basiliximab. We used thymoglobulin in AB0-incompatible patients as induction in the face of high immunologic risk. METHODS: We retrospectively evaluated a cohort of 9 AB0i living donation (LD) recipients from 2011 to 2014. Desensitization included blood group-specific immunoadsorption. Eighteen AB0-compatible LD recipients receiving induction therapy with thymoglobulin served as control subjects. Another control group consisted of 18 AB0-compatible LD recipients receiving basiliximab. Follow-up was 24 months. We captured graft function by estimating glomerular filtration rate (eGFR by Modification of Diet in Renal Disease formula), rejection episodes, and bacterial and viral infections. RESULTS: All patients experienced immediate graft function. No significant or clinical differences were observed regarding graft function, rejection rates, or infections between the groups, although there seemed to be slightly higher cytomegalovirus infection rates due to preemptive therapy strategy. CONCLUSIONS: Thymoglobulin appears to be similar in safety and efficacy to IL-2-antagonists in AB0i kidney transplantation.

Serine phosphorylation site of the 46-kDa mannose 6-phosphate receptor is required for transport to the plasma membrane in Madin-Darby canine kidney and mouse fibroblast cells.

Up to 4% of the human 46-kDa mannose 6-phosphate receptor (MPR46) expressed in Madin-Darby canine kidney (MDCK) cells are localized at the cell surface. At steady state, the expression of MPR46 on the apical surface of filter-grown MDCK cells is about sixfold lower than on the basolateral surface. The cytoplasmic domain of the MPR46 is phosphorylated on serine 56 at low stoichiometry. By expressing mutant MPR46 we have shown that the MPR46 phosphorylation site is required for delivery to the plasma membrane. In addition, mutant MPR46 expressed in MPR-deficient mouse embryonic fibroblasts were not detected at the cell surface and their ability to sort newly synthesized cathepsin D was not altered. Since the loss of MPR46 phosphorylation correlates with the lack of cell surface expression, phosphorylation of serine 56 may either function as a direct plasma membrane targeting signal or inhibit MPR46 recycling from endosomes to Golgi, resulting in trafficking to the cell surface.

First Treatment of Relapsing Rapidly Progressive IgA Nephropathy With Eculizumab After Living Kidney Donation: A Case Report.

BACKGROUND: IgA nephropathy (IGAN) rarely can present as a crescent and progressive form leading to end-stage renal disease (ESRD) in a short period of time. Recurrence of IGAN after kidney transplantation is frequent, and complement components such as C3, C4d, and C5 seem to be involved. We present a case of a young male patient with ESRD caused by rapidly progressive IGAN and who demonstrated rapid recurrence of crescentic IGAN after kidney donation. CASE REPORT: In September 2014, a 28-year-old male patient was hospitalized due to IGAN with 60% of crescents. Cyclophosphamide, steroids, and plasmapheresis did not prevent ESRD. After 8 months of peritoneal dialysis, the patient received a blood group-compatible living donor kidney from his 57-year-old mother. Immunosuppression consisted of tacrolimus, mycophenolic acid, and steroids without induction therapy. Acute graft failure occurred 2 months later, and graft biopsy results revealed recurrence of crescentic IGAN. Cyclophosphamide was added to tacrolimus and steroid treatment, but graft function could not be restored despite viable kidney tissue in repeated biopsy specimens. Rescue therapy with 4 single doses of eculizumab was introduced while hemodialysis had already been initiated. After a cumulative dose of 1800 mg of eculizumab, kidney function did not recover. CONCLUSIONS: In this case, eculizumab was not effective in treating IGAN recurrence after transplantation. Therapy was started late when hemodialysis had already been initiated; an earlier start of therapy might be more effective.

Evaluation of a mass mailing recruitment strategy to obtain a community sample of women for a clinical trial of an incontinence prevention intervention.

OBJECTIVES: Questions exist about using mass mailings to recruit representative samples to participate in clinical trials. The MESA Prevention Study (Medical, Epidemiologic and Social Aspects of Aging), a randomized controlled clinical trial to prevent urinary incontinence (UI), utilized a mass mailing recruitment procedure to recruit a representative sample of women to participate in a behavioral modification program. This paper seeks to expand the literature of mass mailing recruitment strategies for prevention studies by describing the procedures used to recruit healthy, continent, post-menopausal women aged 55-80 years. METHODS: Sociodemographic data collected from recruited subjects is compared with on-line national census data to evaluate the representativeness of the sample recruited from a purchased mailing list. RESULTS: The mass mailing procedure resulted in 3.3% positive response. Of those that returned a positive response, 37.6% were deemed eligible at first screening. Comparisons of study demographic data with state and county census data indicate that the sample obtained was representative of the communities. CONCLUSIONS: The mass mailing strategy was an effective means of recruiting a representative sample of women, aged 55-80. Short falls and recommendations for successful community sample recruitment strategies for clinical trials in older adult women are elaborated upon.

Successful Short-Term Intravenous Treatment of Disseminated Nocardia farcinica Infection with Severe Hyponatremia After Kidney Transplantation: A Case Report.

BACKGROUND: Nocardia is a genus of gram-positive Actinomycetes that are ubiquitous in decaying organic material, soil, and water. Some Nocardia species can infect humans, mainly by airborne transmission. Several reports describe disseminated infections, which are rare and mostly affect strongly immunocompromised patients because intact T-cell-mediated immunity is the major protective mechanism. CASE REPORT: We report a case of disseminated pulmonary, cerebral, and cutaneous infection with Nocardia farcinica in a 66-year-old kidney transplant recipient treated with low-dose triple immunosuppression. The patient was initially admitted because of severe hyponatremia and pneumonia with radiologic signs of pleural effusion. The infectious agent was isolated when cutaneous lesions developed. Oral trimethoprim/sulfamethoxazole treatment led to severe hyponatremia; therefore, long-term treatment with parenteral amikacin and minocycline was initiated. After 7 months of consistent intravenous treatment, the lesions completely resolved and treatment was stopped, against some expert suggestions. The patient had remained free of relapse at the time of writing. CONCLUSIONS: Disseminated Nocardia infection in immunocompromised patients is a rare but life-threatening disease. Owing to its infrequency, the variety of clinical patterns, antimicrobial resistance, and often fatal complications of standardized therapy, the diagnosis and treatment of this infection remain challenging and protracted.

Sexual function in the elderly.

Little is known about sexual behavior among the elderly living in the community. Questions about sexual activity and its correlates were included in a clinic examination whose participants were identified by a household survey of a probability of Washtenaw County, Michigan, elderly, aged 60 years and over, on the medical, epidemiological, and social aspects of aging. Estimates of proportions based on responses at the clinic examination were also projected to the demographics of the household survey. The estimated proportions of individuals who are sexually active are 73.8% for married men and 55.8% for married women; among unmarried men and women the proportions are 31.1% and 5.3%, respectively. The levels decrease significantly with age in both genders. The estimated proportion of married men with erectile impotence is 35.3%. Significant associations were observed between having problems with mobility and the lack of sexual activity in both genders. The prevalence of impotency was significantly associated with a history of heart attack, urinary incontinence, and the use of sedatives. The consumption of at least one cup of coffee per day was significantly associated with a higher prevalence of sexual activity in women and with a higher potency rate in men.

[The use of in-situ hybridization for the detection of Brachyspira spp. in pigs]. / Einsatz der in situ Hybridisierung zum Nachweis von Brachyspira spp. beim Schwein.

Diagnosis of Brachyspira infections in swine and the differentiation of the involved bacteria is time-consuming and in most cases unsatisfactory. Detecting Brachyspira directly in the damaged Brachyspira of the large intestine could provide a direct correlation between histological lesionsa and bacterial growth. In this study we investigated whether in-situ hybridization (ISH) with a digoxigenin-labeled RNA-probe is a suitable method for detecting Brachyspira in the mucosa of the large intestine. Formalin-fixed and paraffin-embedded tissue sections of the large intestine from 78 pigs, which showed macroscopic and histological findings of Brachyspira-associated colitis, were stained with hematoxylin and eosin and Warthin-Starry silver impregnation and subjected to ISH. We used a RNA-probe with a length of 334bp, complementary to a part of the 23S rRNA of all members of the genus Brachyspira. All sections were treated with this anti-sense probe and with a sense control probe. 64 samples (82%) showed clearly positive ISH signals. Thus ISH is a suitable method for detecting Brachyspira directly within the lesions of the large intestine. The quantity of Brachyspira identified by ISH was always lower than by Warthin-Starry staining. Whether this reflects lower sensitivity of the ISH technique, or the fact that other bacteria with morphological similarities to Brachyspira were also stained by Warthin-Starry is unknown as yet. The present investigations provide a basis of further research developing specific probes to distinguish between pathogenic and non pathogenic Brachyspira species and probes detecting other bacteria with morphological similarity to Brachyspira.

The effect of phenytoin and carbamazepine on serum dehydroepiandrosterone sulfate in men and women who have partial seizures with temporal lobe involvement.

In a study of 16 men and 55 women with temporal lobe epilepsy (TLE), we found that the serum dehydroepiandrosterone sulfate (DHEA-S) level was significantly lower (P less than 0.05) in patients who were treated with phenytoin (mean +/- SD DHEA-S in men, 685 +/- 429 ng/ml; in women, 580 +/- 289), carbamazepine (women, 779 +/- 529), or a combination of the two drugs (men, 491 +/- 246; women, 474 +/- 284) than in age- and sex-matched untreated patients (men, 2634 +/- 418; women, 2011 +/- 1435) or in normal subjects (men, 2870 +/- 1052; women, 1764 +/- 617). DHEA-S values in the treated groups did not differ from one another. The DHEA-S levels in untreated patients with TLE did not differ significantly from those in normal subjects. TLE alone, therefore, is unlikely to account for the decrease in DHEA-S, and thus, the results suggest that the antiseizure medications are capable of reducing DHEA-S levels. These findings are important because DHEA-S levels are often measured as part of a battery of tests to assess endocrine function in conditions such as polycystic ovarian syndrome, which may be over-represented among women with TLE.

Polygraphic sleep criteria as predictors of successful aging: an exploratory longitudinal study.

BACKGROUND: A cohort of 57 elderly healthy volunteers (34 male, 23 female) was studied in a sleep laboratory on four consecutive nights when their average age was 63.5 +/- 3.7 years. Thirty subjects (20 male, 10 female) were assessed 14 years later; 21 had either died in the meantime or were very ill, and 6 did not participate for other reasons. METHODS: Two operationalizations of successful aging were applied: survival in relatively good health (30 survivors vs. 21 nonsurvivors), and cognitive competence as assessed in the survivors by means of tests of cognitive function. RESULTS: Whereas none of the sleep characteristics determined at baseline distinguished the survivors from the nonsurvivors, several parameters [REM (rapid eye movement) sleep latency, REM density, and NREM (non-REM) shifts] were significantly correlated with one or more measures of cognitive functioning at follow-up. These polygraphic sleep parameters also distinguished a subgroup of cognitively fully competent subjects from those who, according to their performance in tests of cognitive function, could be considered as mildly demented. CONCLUSIONS: While the REM latency and density findings support the theory of a functional link between brain cholinergic activity, timing, and density of REM sleep and cognitive functioning, the positive association between the number of NREM shifts at baseline and cognitive performance 14 years later is difficult to explain. It is suggested that the findings of the present study, in particular the potential predictive value of REM latency and REM density for cognitive functioning in the old, need replication in other subject samples followed for similar time periods.

Late-onset congenital adrenal hyperplasia: a treatable cause of anxiety.

BACKGROUND: Some intermediaries of cortisol synthesis, especially the sulfated ester of dehydroepiandrosterone (DHEAS), are picrotoxin-like antagonists of the gamma-aminobutyric acid A (GABA-A) receptor and exert potent anxiogenic effects. We report 5 men and 7 women with refractory anxiety disorders, who had late-onset congenital adrenal hyperplasia (CAH), and in whom interactions between neuroactive steroids and anomalous brain substrates may have participated in the pathophysiology and treatment of anxiety. METHODS: Twelve patients with refractory anxiety disorders as defined by DSM-IV had elevated DHEAS and specific enzyme deficiencies diagnostic of CAH. All were treated with adrenal suppressive therapy using ketoconazole or low (physiologic) dose glucocorticoids. Anxiety was rated by the Tension Scale of the Profile of Mood States (POMS Tension) questionnaire before and during hormonal treatment. RESULTS: Reduction of DHEAS was associated with lower anxiety scores in all twelve cases. POMS Tension scores decreased by 55%. Hormonal treatment, which failed to lower DHEAS, was ineffective. CONCLUSIONS: These findings suggest that late onset CAH can contribute to anxiety disorders and that adrenal suppressive therapy or inhibition of steroidogenesis with ketoconazole may be efficacious as adjuvant therapy.

Expression of human alpha 1-antitrypsin in Escherichia coli.

Complementary DNA coding for human alpha 1-antitrypsin has been placed under the control of the lambda PR promotor carrier by the expression vector pCQV2 [1]. In conditions which allow transcription from this promotor (thermoinactivation of the repressor), Escherichia coli cells harbouring the recombinant plasmid pULB1114 express human alpha 1-antitrypsin (+/- 9000 molecules/cell). The product has a Mr of 44000, corresponding to mature unglycosylated alpha 1-antitrypsin.

Surface plasmon resonance studies prove the interaction of skeletal muscle sarcoplasmic reticular Ca(2+) release channel/ryanodine receptor with calsequestrin.

A high affinity molecular interaction is demonstrated between calsequestrin and the sarcoplasmic reticular Ca(2+) release channel/ryanodine receptor (RyR) by surface plasmon resonance. K(D) values of 92 nM and 102 nM for the phosphorylated and dephosphorylated calsequestrin have been determined, respectively. Phosphorylation of calsequestrin seems not to influence this high affinity interaction, i.e. calsequestrin might always be bound to RyR. However, the phosphorylation state of calsequestrin determines the amount of Ca(2+) released from the lumen. Dephosphorylation of approximately 1% of the phosphorylated calsequestrin could be enough to activate the RyR channel half-maximally, as we have shown previously [Szegedi et al., Biochem. J. 337 (1999) 19].

Characterization of human haptoglobin cDNAs coding for alpha 2FS beta and alpha 1S beta variants.

A human liver library, derived from a heterozygous (Hp2-1) donor, has been used to isolate cDNA clones coding for the haptoglobin (Hp) alpha 1S beta and alpha 2FS beta variants. DNA sequencing has shown that the two variants are identical except for the alpha F duplicated segment in Hp alpha 2FS beta. Four nucleotide changes have been found between the phenotypically different F and S regions of the Hp alpha 2 gene, resulting in an Asp,Lys/Asn,Glu substitution.

Seizure control with clomiphene therapy. A case report.

Clinical seizures ceased during six months of clomiphene citrate therapy in a 36-year-old man who had hypogonadotropic hypogonadism, reproductive dysfunction, and a 16-year history of regular seizures. Seizures recurred during the month that followed the discontinuation of clomiphene therapy. This apparent antiseizure effect of clomiphene is discussed in terms of three possible mechanisms of action: (1) Clomiphene may benefit seizures by raising serum levels of antiseizure medications. (2) Clomiphene may improve seizures by normalizing serum testosterone levels. (3) Clomiphene may act at a cerebral level to lessen seizures. A controlled investigation of the effects of clomiphene on epilepsy in the setting of reproductive dysfunction is suggested.

Amygdaloid changes in aging and dementia.

A quantitative and clincally correlated investigation was carried out to determine the divisional distribution of volumetric and cell-packing density changes in the amygdala in patients with advanced aging and senile dementia. Normal controls show relatively little, but nevertheless significant, age-related change. In contrast, the amygdala undergoes very marked degeneration in patients with senile dementia, which preferentially affects the morphologcially, developmentally, and connectionally older medial, medial central, and cortical nuclei. These findings strengthen the possibility that the amygdala participates in the behavioral changes that occur in senile dementia.

Neuroendocrine dysfunction in temporal lobe epilepsy.

Since medial temporal lobe structures are involved in the modulation of endocrine function and temporal lobe epileptics commonly show personality changes that resemble endocrine disorders, the existence of neuroendocrine dysfunction in temporal lobe epileptics was investigated by comparing the response of serum luteinizing hormone levels with intravenous luteinizing hormone-releasing hormone infusion in patients and normal controls. Five of seven consecutive patients had response curves that fell almost entirely outside of the normal control range, and all seven had either baseline or peak values that were outside of the normal range. These findings suggest that hypothalamic-pituitary control of gonadotropin secretion may be altered among patients with temporal lobe epilepsy.

Reproductive endocrine disorders in women with partial seizures of temporal lobe origin.

Of 50 consecutive women with partial seizures of temporal lobe origin (temporal lobe epilepsy [TLE]) evaluated for reproductive dysfunction, 28 had menstrual problems. Of those, 19 had reproductive endocrine disorders. Polycystic ovarian syndrome and hypogonadotropic hypogonadism occurred significantly more often in women with TLE than in the general female population. Polycystic ovarian syndrome was associated with predominantly left-sided lateralization of interictal epileptic discharges; hypogonadotropic hypogonadism was more commonly found with right-sided discharges. Hyposexuality occurred more often in women with predominantly right-sided interictal epileptic discharges and was associated with low serum luteinizing hormone levels. There are several possible interpretations: epileptic discharges in medial temporal limbic structures may disrupt hypothalamic regulation of pituitary gonadotropin secretion; anovulatory cycles of reproductive endocrine disorders may promote the development of epileptic discharges; and TLE and some associated reproductive endocrine disorders may represent the parallel effects of prenatal factors common to the development of the brain and the reproductive system.