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1.  Escalated handling of young C57BL/6 mice results in altered Morris water maze performance 
The handling of experimental animals prior to experimental interventions is often poorly described, even though it may affect the final functional outcome. This study explores how the use of repeated handling of C57BL/6 mice prior to Morris water maze (MWM) tests can affect the performance.
Methods and materials
The handled animals were subjected to the escalating handling protocol, with the investigator spending 5 min per day per cage for 8 days prior to the MWM test. On the last days of handling, the mice were introduced to water and the concept of a hidden platform. The MWM test consisted of four daily trials for 90 s per day for 4 days with a hidden platform. A probe test was performed 4 days after the last learning trial. Control animals were not handled prior to MWM.
Handling reduced the latency to find the platform on the first 2 days of the MWM tests and reduced thigmotaxis. The mice increased their swim speed and elicited more explorative behavior in the learning trials and to some lesser extent in the probe trials.
The improvement in MWM navigation was most likely due to reduced stress and anxiety regarding the investigator and the test. Handled mice displayed less variability than non-handled mice, suggesting that by using a controlled handling protocol prior to the experiments fewer C57BL/6 mice would be needed to achieve statistically significant differences in studies of learning and spatial memory using MWM.
PMCID: PMC3916711  PMID: 24172203
Handling; mice; Morris water maze
2.  Activation of adenosine receptors improves renal antioxidant status in diabetic Wistar but not SHR rats 
Diabetes and hypertension independently contribute to renal injury, and the major mechanisms involved are increased reactive oxygen species (ROS) bioavailability and renin-angiotensin system (RAS) activation. We investigated the role of adenosine in controlling ROS production and RAS activation associated with renal dysfunction in hypertension and diabetes.
Fourteen days after induction of diabetes with streptozotocin in 12-week-old male Wistar and spontaneously hypertensive (SHR) rats, animals were treated during 7 days with 2-chloroadenosine (CADO group, 5 mg/kg/d), a stable analogue of adenosine, or underwent a sham operation procedure. At the end of the study (day 21), intra-arterial systolic blood pressure (SBP) was measured, and 24-h urine and plasma samples and renal tissue were collected.
CADO treatment decreased the plasma glucose concentration and glucose and protein excretion by more than 30% in both strains. CADO treatment decreased SBP in diabetic SHR rats (143 ± 8 versus 114 ± 4 mmHg, p < 0.05), but not in diabetic Wistar rats. The hypotensive effect of CADO was associated to a ∼70% increase in plasma angiotensinogen (AGT) concentration and a ∼50% decrease in urinary AGT excretion. CADO also caused a decrease in medullary and cortical hydrogen peroxide production of about 40%, which was associated with a proportional increase in glutathione peroxidase (GPx) activity in diabetic Wistar but not in diabetic SHR animals.
These results suggest that activation of adenosine receptors improves renal antioxidant capacity in diabetic Wistar but not SHR rats, although it improves glucose metabolism in both strains. Furthermore, activation of adenosine receptors does not seem to be directly influencing AGT production.
PMCID: PMC3916712  PMID: 24195577
Adenosine; angiotensinogen; hydrogen peroxide; hyperglycemia; kidney; systolic blood pressure
3.  Expression and clinicopathological significance of miR-146a in hepatocellular carcinoma tissues 
Aberrant expression of microRNA-146a (miR-146a) has been found in several classes of cancers. However, its expression and clinicopathological contribution in hepatocellular carcinoma (HCC) has not been fully elucidated.
To explore the clinicopathological significance of the miR-146a level in HCC formalin-fixed paraffin-embedded (FFPE) tissue.
Eighty-five HCC samples and their para-cancerous normal liver tissues were collected. Total mRNA including miRNA was extracted, and miR-146a expression was determined using real-time RT-PCR. Furthermore, the correlation between the miR-146a expression and clinicopathological parameters was investigated.
MicroRNA-146a expression in HCC tissues was lower compared with that in adjacent non-cancerous hepatic tissues. MicroRNA-146a expression was also related to clinical TNM stage, metastasis, portal vein tumor embolus, and number of tumor nodes.
Down-regulation of miR-146a is related to HCC carcinogenesis and deterioration of HCC. MicroRNA-146a may act as a suppressor miRNA of HCC, and it is therefore a potential prognostic biomarker for HCC patients.
PMCID: PMC3916713  PMID: 24172202
Hepatocellular carcinoma; metastasis; miR-146a; oncogenes; paraffin-embedded tissues; pathology; RT-qPCR; tumor biology
4.  Aspirin resistance in patients with type II diabetes mellitus 
Diabetic patients exhibit platelet hyperreactivity, which renders them resistant to antithrombotic treatments. We aimed to investigate the prevalence and predictors of aspirin resistance in diabetic patients.
Material and methods
A total of 93 diabetic and 37 non-diabetic participants were included into the study. Aspirin resistance was measured with a whole-blood desktop platelet function analyzer (PFA-100) with an epinephrine agonist.
Altogether 41.9% patients with DM were aspirin non-responders. Aspirin resistance was observed in 43.2% of non-diabetic patients (p = 0.89). Presence of diabetes mellitus had no effect on aspirin response (RR 0.95 (95% CI 0.44–2.05), p = 0.89) in the whole study population. Hypercholesterolemia was the only predictor of aspirin resistance in multivariate analysis in diabetic patients (RR 3.09 (95% CI 1.17–8.16), p = 0.023).
The prevalence of aspirin resistance is comparable in diabetic and non-diabetic patients. Hypercholesterolemia is the only independent predictor of aspirin resistance in diabetic patients.
PMCID: PMC3916714  PMID: 24236750
Aspirin resistance; diabetes mellitus; PFA-100
5.  Quality of life and functional outcomes 10 years after laparoscopic radical prostatectomy 
Minimally invasive laparoscopic radical prostatectomy (LRP) has proven equally effective as open surgery in terms of cancer control and peroperative complication rate with less bleeding and postoperative pain. However, long-term follow-up data after LRP are scarce, especially as related to quality of life (QoL).
To compare QoL and functional outcomes at least 10 years after LRP with a population-based control group matched for age and region.
Follow-up data were obtained by mailed questionnaires from patients who responded anonymously to five international questionnaires (EQ-5D, QLQ-C30, QLQ-PR25, IPSS, and IIEF). We collected self-reported outcome data directly from 49 patients who underwent LRP more than 10 years ago in our centre. The results of the patients' overall QoL and urinary continence rates were compared with 918 controls matched for region and age.
Forty-two patients (86%) and 808 (88%) controls reported having no urinary leakage. Only 11 patients (24%) still had sexual activities 10 years after LRP, and three were without erectile dysfunction. There was no difference in four of five statements of the self-assessed QoL questionnaires between the LRP and control group. Anxiety level was higher in the LRP group (44%) than in the control group (23%).
Patients reported high self-assessed QoL, although they also reported low sexual activity 10 years after LRP. Prevalence of urinary leakage was similar in both groups. However, anxiety was more common in LRP patients.
PMCID: PMC3916715  PMID: 24328550
Functional outcome; laparoscopic radical prostatectomy; prostate cancer; quality of life
6.  Outcome from spontaneous subarachnoid haemorrhage—results from 2007–2011 and comparison with our previous series 
The management of patients with spontaneous subarachnoid haemorrhage (SAH) has changed, in part due to interventions now being extended to patients who are older and in a worse clinical condition. This study evaluates the effects of these changes on a complete 5-year patient material.
Demographic data and results from 615 patients with SAH admitted from 2007 to 2011 were put together. Aneurysms were found in 448 patients (72.8%). They were compared with the aneurysm group (n = 676) from a previously published series from our centre (2001–2006). Linear regression was used to determine variables predicting functional outcome in the whole aneurysm group (2001–2011).
Patients in the more recent aneurysm group were older, and they were in a worse clinical condition on admission. Regarding younger patients admitted in World Federation of Neurosurgical Societies SAH grading (WFNS) 3, there were fewer with a good outcome. In the whole aneurysm group 2001–2011, outcome was best predicted by age, clinical condition at admission, and the size of the bleeding, and not by treatment mode or localization of aneurysm.
It seems important for the outcome that aneurysms are treated early. The clinical course after that depends largely on the condition of the patient on admission rather than on aneurysm treatment method. This, together with the fact that older patients and those in worse condition are now being admitted, increases demands on neurointensive care. Further improvement in patient outcome depends on better understanding of acute brain injury mechanisms and improved neurointensive care as well as rehabilitation measures.
PMCID: PMC3916716  PMID: 24147458
Aneurysm; endovascular; outcome; neurointensive care; subarachnoid haemorrhage; surgery
7.  Recurrent pulmonary aspergillosis and mycobacterial infection in an unsplenectomized patient with type 1 Gaucher disease 
The clinical presentation of Gaucher disease (GD), an inherited lysosomal storage disorder caused by the deficient activity of the lysosomal enzyme glucocerebrosidase, is highly variable, and three clinical types are distinguished based upon the presence of neurologic symptoms. Thrombocytopenia, anemia, hepatosplenomegaly, and bone manifestations are the most typical signs of GD type 1 (GD1).
Case presentation
We present the case of an unsplenectomized man suffering from heterozygous GD1 with mutations of c.1226A>G (N370S) and RecNci I (L444P, A456P, and V460V) in the GBA1 gene, who developed recurrent pulmonary aspergillosis caused by Aspergillus fumigatus and a mycobacterial infection caused by Mycobacterium avium. Despite long-lasting therapy of both aspergillosis (including antifungal drugs and surgery), and the mycobacterial infection (triple therapy with rifampicin, ethambutol, and clarithromycin), recurrent positivity for M. avium and A. fumigatus was detected.
Symptomatic lung involvement and an increased susceptibility to pulmonary infections are uncommon in GD and, if present, are often associated with more severe disease manifestations. To our knowledge, this is the first published report on the association of GD and pulmonary aspergillosis and mycobacterial infection. It illustrates the increased susceptibility of untreated GD patients to opportunistic pulmonary infections and ineffective eradication of these infections despite adequate therapy.
PMCID: PMC3916717  PMID: 24195576
Aspergillosis; Aspergillus fumigatus; Gaucher disease; mycobacterial infection; Mycobacterium avium
8.  An uncommon cause of acutely altered mental status in a renal transplant recipient 
Neurological complications are quite frequent in patients after solid organ transplantation presenting with focal or generalized neurologic symptoms as well as altered mental status. Posterior reversible encephalopathy syndrome is a rare cliniconeuroradiological entity characterized by headache, altered mental status, cortical blindness, seizures, and other focal neurological signs and a diagnostic magnetic resonance imaging.
Case report
We present a case of a 57-year-old woman with one episode of seizures and sudden onset of altered mental status (time and person perception) accompanied with headache at the thirtieth postoperative day after renal transplantation.
Posterior reversible encephalopathy syndrome, although an uncommon post-renal transplantation complication, should be considered in these patients, as several factors surrounding the setting of transplantation have been implicated in its development. Thus, physicians should be aware of this condition in order to establish the diagnosis and offer appropriate treatment.
PMCID: PMC3916718  PMID: 24079344
Depression; end-stage renal disease; hypertension; posterior reversible encephalopathy syndrome; tacrolimus
13.  Serum pentraxin 3 concentrations in neonates 
PMCID: PMC3916723  PMID: 24438192
14.  Tumor length in elderly patients with esophageal squamous cell carcinoma: Is it a prognostic factor? 
Upsala Journal of Medical Sciences  2013;118(3):145-152.
Several researchers have determined the tumor length to be an important prognostic indictor of esophageal cancer (EC). However, controversy exists concerning the optimal cut-off points for tumor length to predict overall survival. The aim of this study was to determine the prognostic value of tumor length and propose the optimum cut-off point for tumor length in predicting survival difference in elderly patients with esophageal squamous cell carcinoma (ESCC).
From January 2001 to December 2009, a retrospective analysis of 132 consecutive patients older than 70 years with ESCC was conducted. A receiver-operating characteristic (ROC) curve for survival prediction was plotted to verify the optimum cut-off point for tumor length. Univariate and multivariate analyses were performed to evaluate prognostic parameters for survival.
A ROC curve for survival prediction was plotted to verify the optimum cut-off point for tumor length, which was 4.0 cm. Patients with tumor length ≤4.0 cm had significantly better 5-year survival rate than patients with a tumor length >4.0 cm (60.7% versus 31.6%, P = 0.007). Multivariate analyses showed that tumor length (>4.0 cm versus ≤4.0 cm, P = 0.036), differentiation (poor versus well/moderate, P = 0.032), N staging (N1-3 versus N0, P = 0.018), and T grade (T3-4 versus T1-2, P = 0.002) were independent prognostic factors.
Tumor length is a predictive factor for long-term survival in elderly patients with ESCC, especially in T3-4 grade or nodal-negative patients. We conclude that 4.0 cm may be the optimum cut-off point for tumor length in predicting survival in elderly patients with ESCC.
PMCID: PMC3713379  PMID: 23617771
Esophageal cancer; esophagectomy; prognostic factor; squamous cell carcinoma; survival; tumor length
15.  Dynamic changes in glycosylation and glycan composition of serum FSH and LH during natural ovarian stimulation 
Upsala Journal of Medical Sciences  2013;118(3):153-164.
Glycosylation and glycan composition are of fundamental importance for the biological properties of FSH and LH. The aim of this study was to determine the glycosylation, sialylation, and sulfonation of serum FSH and LH throughout the normal menstrual cycle.
Serum samples were collected from 79 healthy women with regular menstrual cycles. The mean numbers of anionic monosaccharide (AMS), sialic acid (SA), and sulfonated N-acetylgalactosamine (SU) residues per FSH and LH molecule were estimated for all sera with methods based on electrophoreses, neuraminidase treatments, and fluoroimmunoassays of the gonadotrophins.
Di-glycosylated glycoforms (FSHdi, LHdi) were detected in serum in addition to tetra-glycosylated FSH (FSHtetra) and tri-glycosylated LH (LHtri). FSHdi exhibited two peaks: one on day 5 to 7 and one, more pronounced, at midcycle. FSHtetra plateaued at a high concentration from day 5 to 15, without a midcycle peak. There were lower concentrations of LHdi than LHtri, except at midcycle when the opposite occurred. The mean numbers of SA and SU residues per molecule of FSH and LH in serum showed four different patterns during the cycle, all with highly significant (P < 0.0001) differences between levels at different phases of the cycle. The pattern of SA residues on FSH was ‘M’-shaped, and that of SU on LH ‘V’-shaped.
Serum FSH and LH governing the natural ovarian stimulation process exhibited dynamic changes of glycosylation and glycan composition. This new information on the FSH and LH molecular structures may lead to more successful mono-ovulatory treatment regimens for ovulation induction in anovulatory women.
PMCID: PMC3713380  PMID: 23527482
di-glycosylated FSH; di-glycosylated LH; glycoforms; isoforms; ovulation induction; sialic acid; sulfonated N-acetylgalactosamine
16.  Lifestyle intervention is associated with decreased concentrations of circulating pentraxin 3 independent of CRP decrease 
Upsala Journal of Medical Sciences  2013;118(3):165-168.
Pentraxin 3 (PTX3) is an acute phase marker, which is produced at the site of infection or inflammation in contrast to CRP that is mainly synthesized by the liver. The aim of the present study was to see if lifestyle interventions/weight loss would lead to decreased blood plasma concentrations of PTX3.
Study subjects (n = 31) were recruited to a lifestyle intervention program aiming at increased physical activity, improved eating habits, and weight loss. High-sensitivity C-reactive protein (CRP) and PTX3 methods were used for analysis of CRP and PTX3 in plasma samples collected at inclusion and after 4 and 8 weeks of treatment.
Wilcoxon paired samples test showed a significant decrease in PTX3 concentrations from 2068 pg/mL at start to 2007 pg/mL at 4 weeks (P = 0.002) and 1748 pg/mL at 8 weeks (P = 0.003). The PTX3 decrease was not significantly correlated with a corresponding decrease in CRP or weight reduction.
The lifestyle intervention program resulted in a significant reduction of circulating concentrations of pentraxin 3 already after 4 and 8 weeks of treatment.
PMCID: PMC3713381  PMID: 23837595
Adult; body weight; CRP; human; inflammation; pentraxin 3; plasma
17.  Introduction of the Uppsala Traumatic Brain Injury register for regular surveillance of patient characteristics and neurointensive care management including secondary insult quantification and clinical outcome 
Upsala Journal of Medical Sciences  2013;118(3):169-180.
To improve neurointensive care (NIC) and outcome for traumatic brain injury (TBI) patients it is crucial to define and monitor indexes of the quality of patient care. With this purpose we established the web-based Uppsala TBI register in 2008. In this study we will describe and analyze the data collected during the first three years of this project.
Data from the medical charts were organized in three columns containing: 1) Admission data; 2) Data from the NIC period including neurosurgery, type of monitoring, treatment, complications, neurological condition at discharge, and the amount of secondary insults; 3) Outcome six months after injury. Indexes of the quality of care implemented include: 1) Index of improvement; 2) Index of change; 3) The percentages of ‘Talk and die' and ‘Talk and deteriorate' patients.
Altogether 314 patients were included 2008–2010: 66 women and 248 men aged 0–86 years. Automatic reports showed that the proportion of patients improving during NIC varied between 80% and 60%. The percentage of deteriorated patients was less than 10%. The percentage of Talk and die/Talk and deteriorate cases was <1%. The mean Glasgow Coma Score (Motor) improved from 5.04 to 5.68 during the NIC unit stay. The occurrences of secondary insults were less than 5% of good monitoring time for intracranial pressure (ICP) >25 mmHg, cerebral perfusion pressure (CPP) <50 mmHg, and systolic blood pressure <100 mmHg. Favorable outcome was achieved by 64% of adults.
The Uppsala TBI register enables the routine monitoring of NIC quality indexes.
PMCID: PMC3713382  PMID: 23837596
Database; neurointensive care; outcome; quality register; secondary insults; traumatic brain injury
18.  Palliative care in COPD—web survey in Sweden highlights the current situation for a vulnerable group of patients 
Upsala Journal of Medical Sciences  2013;118(3):181-186.
Chronic obstructive pulmonary disease (COPD) is a common cause of death. Despite the heavy symptom burden in late stages, these patients are relatively seldom referred to specialist palliative care.
A web-based survey concerning medical and organizational aspects of palliative care in COPD was distributed to respiratory physicians in Sweden. There were 93 respondents included in the study.
Palliative care issues were regularly discussed with the patients according to a third of the respondents. About half of the respondents worked in settings where established routines for co-operation with palliative units were available at least to some extent. Less than half of the respondents (39%) were aware of current plans to develop palliative care, either as a co-operative effort or within the facility. Palliative care is focused on physical, psychological, social, and existential dimensions, and the proportions of respondents providing support within these dimensions, ‘always' or ‘often', were 83%, 36%, 32%, and 11%, respectively. Thus, to treat the physical dimensions was perceived as much more obvious than to address the other dimensions.
The survey indicates that the priorities and resources for palliative care in COPD are insufficient in Sweden. The data, despite limitations, reveal a lack of established team-work with specialized palliative care units and actual plans for such co-operation.
PMCID: PMC3713383  PMID: 23710665
Care management; COPD; holistic care; palliative care; survey
19.  Attitudes towards embryo donation in Swedish women and men of reproductive age 
Upsala Journal of Medical Sciences  2013;118(3):187-195.
When performing in-vitro fertilization (IVF), more embryos than needed are often derived. These embryos are usually frozen and stored, but as ruled by Swedish law they have to be discarded after 5 years. In other countries it is legal to donate the excess embryos to other infertile couples who for different reasons cannot undergo the procedure of IVF. The aim of the present study was to investigate public opinion in Sweden regarding different aspects of embryo donation.
A questionnaire regarding attitudes towards aspects of embryo donation was sent to a randomized sample of 1,000 Swedish women and men of reproductive age.
A total of 34% responded to the questionnaires. A majority of the respondents (73%) were positive towards embryo donation. Seventy-five per cent agreed that it should be possible to donate embryos to infertile couples. Approximately half of the participants (49%) supported embryo donation to single women. A majority of the participants emphasized that demands should be imposed on the recipient's age (63%), alcohol addiction (79%), drug addiction (85%), and criminal record (67%). Forty-seven per cent of the respondents agreed that the recipient should be anonymous to the donor, and 38% thought that the donor should remain anonymous to the child.
The results of the present study indicate support for embryo donation among a subset of the Swedish population of reproductive age. If embryo donation were to be allowed in Sweden, strategies for treatment and counselling need to be developed.
PMCID: PMC3713384  PMID: 23786323
Attitudes; disclosure; embryo donation; gender; Swedish population
20.  Neoplastic pericarditis as the initial manifestation of a papillary thyroid carcinoma 
Upsala Journal of Medical Sciences  2013;118(3):196-198.
Neoplastic pericarditis represents approximately 5%–7% of the cases with acute pericarditis and is rarely the initial manifestation of malignancy. The most common cause is lung cancer, followed by breast cancer, lymphomas, leukemia, and esophageal cancer. Neoplastic pericardial disease is extremely rare in thyroid cancer, especially as the first manifestation. Here, we present a papillary thyroid carcinoma that was manifested with pericarditis and cardiac tamponade in a 49-year-old female.
PMCID: PMC3713385  PMID: 23697667
Cardiac tamponade; neoplastic pericarditis; pericardial effusion; thyroid cancer
22.  Hemophagocytic lymphohistiocytosis in adults 
Upsala Journal of Medical Sciences  2013;118(3):201-203.
PMCID: PMC3713387  PMID: 23682633

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