A new hybrid approach for MHC genotyping: high-throughput NGS and long read MinION nanopore sequencing, with application to the non-model vertebrate Alpine chamois (Rupicapra rupicapra).

The major histocompatibility complex (MHC) acts as an interface between the immune system and infectious diseases. Accurate characterization and genotyping of the extremely variable MHC loci are challenging especially without a reference sequence. We designed a combination of long-range PCR, Illumina short-reads, and Oxford Nanopore MinION long-reads approaches to capture the genetic variation of the MHC II DRB locus in an Italian population of the Alpine chamois (Rupicapra rupicapra). We utilized long-range PCR to generate a 9 Kb fragment of the DRB locus. Amplicons from six different individuals were fragmented, tagged, and simultaneously sequenced with Illumina MiSeq. One of these amplicons was sequenced with the MinION device, which produced long reads covering the entire amplified fragment. A pipeline that combines short and long reads resolved several short tandem repeats and homopolymers and produced a de novo reference, which was then used to map and genotype the short reads from all individuals. The assembled DRB locus showed a high level of polymorphism and the presence of a recombination breakpoint. Our results suggest that an amplicon-based NGS approach coupled with single-molecule MinION nanopore sequencing can efficiently achieve both the assembly and the genotyping of complex genomic regions in multiple individuals in the absence of a reference sequence.

Hyponatremia is a predictor of hospital length and cost of stay and outcome in cancer patients.

PURPOSE: Hyponatremia is the most common electrolyte disorder in hospitalized patients, and it might be an indicator of poor prognosis and might have negative effects on hospitalization length and quality of life in non-malignant as well as in malignant diseases. The aim of this study is to determine the impact of hyponatremia on the length and on the cost of hospitalization as well as on outcome in cancer patients. METHODS: The present study includes 105 consecutive cancer patients hospitalized at our institution from June 2013 to December 2013. Data regarding age, sex, staging, histology, chemotherapy, and serum sodium levels at admission, during hospitalization, and at discharge were recorded and statistically analyzed. Impact of hyponatremia on length and cost of hospitalization and on outcome was evaluated. RESULTS: A significant difference in overall survival since the date of admission was observed between eunatremic and hyponatremic patients (p = 0.0255). A statistically significant correlation was also found between the length of stay and the detection of hyponatremia. At multivariate analysis, hyponatremia at admission, severity of hyponatremia, and stage of disease resulted independent prognostic factors. Furthermore, a patient with moderate or severe hyponatremia cost, in rate terms, 128 and 299 % more than a normonatremic patient, respectively. CONCLUSIONS: The occurrence of hyponatremia at the admission or during the hospitalization may represent a significant factor influencing the outcome and the length of hospitalization. Acting effective and timely on the normalization of sodium levels might have a positive effect on prognosis in this setting of patients, as well as on the length of stay in hospital, thus potentially resulting in savings.

A possible role of atrial fibrillation as a risk factor for dementia.

Atrial fibrillation (AF), which is a very common disease among the elderly, is already well known as a risk factor for arterial thromboembolism and stroke. The attention of medical research is now focused on establishing a possible role of AF in the development of cognitive impairment in order to include this arrhythmia among risk factors for dementia. The aim of this work was to investigate the relationship between AF and various types of dementia, such as vascular dementia (VaD), Alzheimer's disease (AD) and mixed dementia (MD). The study consisted of 71 VaD, AD or MD patients, 31 males and 40 females. The sample has been divided in 2 groups according to the sex, and these two groups have been analyzed separately. In females, a statistically significant association was found between mini mental state examination (MMSE) and clinical dementia rating (CDR) scores and AF occurrence (r=-0.32; p<0.05; r=0.33; p<0.05). On the contrary, no significant linear correlation was found between AF and a lower activities if daily living (ADL) and instrumental activities if daily living (IADL) scores. In males, AF/MMSE, AF/CDR, AF/ADL and AF/IADL variables have not been found to be linearly related to each other. Unexpectedly, AF turned to be associated to AD more often than to VAD, becoming a possible risk factor for this neurodegenerative disease. Our results are supported by many studies in literature attributing a basic role of brain hypoperfusion in sporadic AD patho-genesis. More and more scientific data suggest that the already well known risk factors for AD could be considered just the top of an iceberg, providing powerful arguments for impaired cerebral perfusion as the primary trigger in the development of this disease. Moreover, the mildly favorable treatment response in patients with AD to therapy that improves cerebral blood flow is a consistent finding; the same cannot be said of antiamyloid treatments. This opens new possibilities to find an effective way to treat this dramatic pathology.

Acute respiratory distress syndrome secondary to antisynthetase syndrome is reversible with tacrolimus.

Polymyositis and interstitial lung diseases, predominantly nonspecific interstitial pneumonia (NSIP), are known to be frequent in antisynthetase syndrome, where anti-aminoacyl-tRNA synthetase antibodies are often identified. An unusual case of acute respiratory distress syndrome, secondary to such proven NSIP of cellular type with predominant CD8 lymphocytes, is described herein. The patient described in the present case study initially had a poor recovery with high dose of steroids, but this was followed by a good improvement after the prescription of tacrolimus and a low dose of prednisone. A precise diagnosis in similar circumstances may be life-saving, allowing the successful application of new immunosuppressants.

Improvement in the quality of the catering service of a rehabilitation hospital.

BACKGROUND: Malnutrition due to undernutrition or overnutrition is highly prevalent in hospital in-patients and it decisively conditions patients clinical outcome. One of the most influencing factors of malnutrition in hospitalized patients is--at least in part--the Catering Service Quality. AIM: Is to verify, over a 5 year period, the course of the quality of the institutional Catering Service, verifying the effectiveness of the quality improvement process used. METHODS: Quality control was performed by objective (meal order accuracy, proper distribution of food in trolleys, route time from the kitchen to the ward and time of food distribution, food weight and temperature, waste assessment) and subjective assessment (quality was measured by giving the patients a questionnaire after meals). RESULTS: The survey included: 572 meals and 591 interviews. A significant amount of "qualitative" errors (lack of respect for patient preferences or at the moment of supplying the food trolley) have been found. Over the time and the amount of patients that wasted a considerable amount of the portion served was considerably reduced food temperature have been improved. Also patient satisfaction with menu variability, portion size, temperature and cooking quality improved over time. The overall ratings of meals under observation improved too in fact, positive opinions ranged from 18% in 2002 to 48.3% in 2006. CONCLUSION: Ongoing research and quality verification, which include all catering service workers, yields a constant improvement in quality. Patients in healthcare settings should receive a service they appreciates, but it should be--at the same time--correct from a nutritional point of view. For this reason, it is necessary a continuous mediation between customers satisfaction and nutritionists work, dieticians and nursing staff. From this point of view the educational approach becomes essential to feed patient compliance to dietetic treatment that will continue after discharge.

Administrative monitoring of tuberculosis treatment in Switzerland.

SETTING: Treatment of tuberculosis (TB) is critically dependent on adherence. Directly observed treatment (DOT) has been shown to be effective. OBJECTIVE: To determine operational treatment outcome using administrative treatment monitoring (ATM) to assess the need for more vigorous promotion of DOT. DESIGN: Cohort study in eastern Switzerland, where ATM was started in 2002. Bi-monthly progress forms and a treatment outcome form (after 6 months) were sent to the treating doctors. Forms not returned within 6 weeks were followed up with phone calls. RESULTS: Between 2002 and 2004, 98 (87.5%) of 112 new TB patients completed a 6-month treatment course. Eight elderly patients died of causes other than TB while on treatment, four travelled out of the region and two were lost to follow-up. Treating doctors opted for DOT in only seven cases. CONCLUSION: Given the high success rate of 87.5% in our cohort, more vigorous promotion of DOT is not a priority for TB case management in eastern Switzerland. In our setting, ATM in collaboration with the family doctors offers a valuable alternative to the more time-consuming universal DOT.

Correlation between cognitive impairment and the Rey auditory-verbal learning test in a population with Alzheimer disease.

Patients affected by Alzheimer disease (AD) need an accurate diagnosis, to the extent allowing us to find the best therapy or polytherapy, in order to take under control their cognitive impairment. In our Alzheimer Evaluation Units (from the Italian name abbreviated: UVA), the patients undergo a multidimensional evaluation, which can address us towards a proper diagnosis and of other weakening, or even dementia-related diseases. The patients are also subject to neuropsychometric and neuropsychological evaluations, allowing a more focused analysis on cognitive impairments. Among the tests, we use the Rey auditory-verbal learning test (RAVLT), evaluating the patient's verbal memory. A list of 15 words is read to each patient. N the first part of the test, the clinician repeats 5 times such a list. the patient is hen asked, at the end of every repetition, to tell all words he/she remembers. This part is useful to evaluate the immediate recall (IR) ability. The score, i.e., the total number of recalled words, ranges from 0 to 75. After 15 minutes, the delayed recall (DR) ability is evaluated: the patient is newly asked to repeat as many words as he can recall from the list. The score for this part ranges from 0 to 15 minutes. The score is corrected of rage and education, with a cut-off of 28.5 for IR and 4.7 for DR. We made a survey with the purpose of deciding if there was a correlation between cognitive impairment and verbal memory lack, whose deficiency appears earlier in AD. To this aim, we selected several patients with AD, diagnosed during the period between September 2002 and February 2003. We only considered those patients whose AD was not associated with other weakening diseases, and whose clinical dementia rating scale (CDR) score was between 0.5-2.0. A sample of 35 individuals (11 men and 24 women) could be obtained. A meaningful correlation was observed between CDR and IR (r = -0.725, p < 0.01), as well as between CDR and DR (r = -0.470; p < 0.05). Such a result confirms the importance of evaluating immediate and long-term memories, for the early diagnosis of AD, because it is the only symptom of clinically not yet diagnosed dementia, as proven also by other studies.

Gastric emptying time in cyclic vomiting syndrome in children.

Pathogenesis of cyclic vomiting syndrome (CVS) is still uncertain. Aim of our study has been to verify the role of gastric emptying time in children affected by CVS. We studied 9 children with CVS who underwent measurement of gastric emptying time by real-time ultrasonography after administration of a mixed, solid-liquid meal. In all the children gastric emptying time resulted in normal range. Our data suggest that mobility abnormalities of the stomach don't play a key role in the pathogenesis of CVS.

[From social distress to psychic pain]. / De la détresse sociale à la souffrance psychique.

Regarding 75 subjects of both sex sent to psychiatric and alcological consultations by social departments to which they required some help, the authors ask the question about limits between social alienation and mental alienation for people in precarity and poverty position. They inquire into role part and position of the psychiatrist beside them.

Controlled trial of low calcium versus high calcium intake in mild hypertension.

A controlled trial of the effect of low versus high calcium intake on blood pressure was performed in 15 patients with mild essential hypertension (supine blood pressure after a 1-month run-in period: 145.7 +/- 2.6/97.8 +/- 0.9 mmHg, mean +/- s.e.m.). After a 1-week baseline period on a standard calcium intake (900 mg/day, obtained by giving a 500-mg calcium tablet daily, in addition to a 400-mg calcium diet), the patients were randomly entered into a double-blind crossover study of 4-week low calcium intake (400 mg calcium diet plus two placebo tablets/day) and 4-week high calcium intake (1400 mg/day: 400-mg calcium diet plus two 500-mg calcium tablets/day). Compliance with the diets appeared to be satisfactory, based on the results of food record analysis. No significant blood pressure change was observed at the end of the low-compared to the high-calcium regimen. Serum ionized calcium was slightly, but not significantly lower, while 24-h urinary calcium excretion was significantly reduced during the low-calcium diet. No difference was found in urinary sodium and potassium excretion between the two study periods. We conclude that moderate modifications of oral calcium intake are not associated with changes in blood pressure within the time span of this study.

Controlled trial of long term oral potassium supplements in patients with mild hypertension.

A 15 week randomised double blind placebo controlled trial of oral potassium supplements (48 mmol daily) was conducted in 37 patients who had mildly increased blood pressure and a normal dietary intake of sodium. After a two month run in and a one week baseline period the patients were randomly assigned to receive either potassium supplements (n = 18) or placebo (n = 19). By the third week of treatment blood pressure in the actively treated group had decreased significantly compared with that in the placebo group, though the decrease reached its maximum after 15 weeks. Urinary potassium excretion increased significantly in the group who received potassium supplements, but no significant changes were found in plasma sodium and potassium concentrations or in urinary sodium excretion. In a subgroup of 13 patients who underwent a further nine weeks of treatment with oral potassium supplements at half of the previous dose (24 mmol daily) their blood pressure, at the end of this second study period, was still significantly lower compared with their baseline value but not with that of the placebo group. These results show that moderate oral potassium supplements are associated with a long term reduction in blood pressure in patients who have mild hypertension.