bluuebunny/crossref_metadata_2025 huggingface dataset:
Multidisciplinary assessment of tako tsubo cardiomyopathy: a prospective case study
Authors: Waldenborg Micael , Soholat Mona , Kähäri Anders , Emilsson Kent , Fröbert Ole
Published: April 2011

Background

The cause of tako tsubo cardiomyopathy remains unclear. We used a multidisciplinary approach to investigate if a common pathophysiological denominator could be outlined.

Methods

Within 3 days following symptom presentation and again after 3 months we investigated all patients coming to our institution and diagnosed with tako-tsubo cardiomyopathy. Patients underwent extensive biochemical screening. Left ventricular function was evaluated by echocardiography and contrast-enhanced cardiac magnetic resonance imaging. Cardiac autonomic function was studied by heart rate variability and signal-averaged electrocardiogram and posttraumatic stress and depression were investigated by questionnaires (the Posttraumatic Stress Syndrome 10-Questions Inventory, PTSS-10 and the Montgomery-Åsberg depression rating scale, self rated version, MADRS-S).

Results

During 2 years, 13 consecutive patients were included. Markers of myocardial damage and heart failure were slightly to moderately elevated and ejection fraction (echocardiography and MRi) was moderately reduced at hospitalization and improved to normal values in all patients. Signal averaged ECG demonstrated a statistically significant shorter duration of the filtered QRS complex in the acute phase as compared to follow-up. In heart rate variability analysis, SDNN and SDANN were shorter acutely compared to follow-up. Two patients fulfilled criteria for posttraumatic stress syndrome while 7 patients were in the borderline zone. There was a statistically significant inverse correlation between PTSS-10 score and QRS duration in the signal-averaged ECG (r = -0.66, P = 0.01).

Conclusions

Patients with tako tsubo cardiomyopathy have altered cardiac autonomic function and a high incidence rate of borderline or definite posttraumatic stress syndrome acutely. This is in line with findings in patients with myocardial infarction and does not allow conclusions on cause and effect.
R353Q polymorphism in the factor VII gene and cardiovascular risk in Heterozygous Familial Hypercholesterolemia: a case-control study
Authors: Criado-García Juan , Fuentes Francisco , Cruz-Teno Cristina , García-Rios Antonio , Jiménez-Morales Anabel , Delgado-Lista Javier , Mata Pedro , Alonso Rodrigo , López-Miranda José , Pérez-Jiménez Francisco ,   Spanish Group for the Study of Familiar Hypercholesterolemia
Published: April 2011

Background

Heterozygous Familial Hypercholesterolemia (FH) is a genetic disorder characterized by a high risk of cardiovascular disease. Certain polymorphisms of the factor VII gene have been associated with the development of coronary artery disease and there is a known association between factor VII levels and polymorphic variants in this gene. To date, no study has evaluated the association between factor VII and coronary artery disease in patients with FH.

Results

This case-control study comprised 720 patients (546 with FH and 174 controls). We determined the prevalence and allele frequencies of the R353Q polymorphism of factor VII, the plasma levels of factor VII antigen (FVII Ag) and whether they could be predictive factors for cardiovascular risk. 75% (410) of the patients with FH were RR, 23% (127) RQ and 1.6% (9) QQ; in the control group 75.3% (131) were RR, 21.3% (37) RQ and 3.4% (6) QQ (p = 0.32). No statistically significant associations were observed in the distribution of genotypes and allele frequencies between case (FH) and control groups. Nor did we find differences when we evaluated the relationship between the R353Q polymorphism and cardiovascular risk (including coronary disease, ischemic stroke and peripheral arterial disease), either in the univariate analysis or after adjustment for sex, age, arterial hypertension, body mass index, xanthomas, diabetes, smoking, HDLc and LDLc and lipid-lowering treatment. The FVII Ag concentrations behaved in a similar fashion, with no differences for the interaction between controls and those with FH (RR vs. RQ/QQ; p = 0.96). In the subgroup of patients with FH no association was found among cardiovascular disease, genotype and FVII Ag levels (RR vs. RQ/QQ; p = 0.97).

Conclusions

Our study did not find a direct relationship between cardiovascular risk in patients with Heterozygous Familial Hypercholesterolemia, the R353Q polymorphism of factor VII and FVII Ag levels.
Transnational telepathology consultations using a basic digital microscope: experience in the Italy-Slovenjia INTERREG project “Patient without borders”
Authors: Intersimone Donatella , Snoj Viviana , Riosa Franca , Bortolotti Nicola , Sverko Sandi , Beltrami Carlo A , Della Mea Vincenzo
Published: April 2011

Background

In recent years, a number of technological advancements started to modify the long standing appearance and functionalities of traditional optical microscopes used in Pathology and other medical fields. In fact, at present many new tools for microscopical visualization exist that are based on digital imaging, robotization, and remote communication. Such tools are typically adopted in activities ranging from education to telediagnosis to remote consultation.

Present paper describes the features of a basic digital microscope that has been tested to verify its performance for occasional remote consultation inside an international project between Italy and Slovenija, funded by Interreg initiative of the European Regional Development Fund.

Methods

The system is composed by a pair of digital microscopes (Leica DMD108, Leitz Microsystems, Wetzlar, Germany) associated to a high resolution videoconferencing systems (Tandberg 990, Lysaker, Norway). The systems are connected through the Internet. Sixty histology and cytology cases have been collaboratively diagnosed between two Pathology Institutes to verify the diagnostic performance of the system, regarding the image quality point of view as well as time needed for diagnosis. The system has also been tested for compatibility with standard videoconferencing software.

Results

No discrepancies between local and remote diagnoses have been identified, with diagnosis time reasonably close to typical microscope observation times. Time needed for most operations is not far from that needed on a traditional microscope, except for startup.

Conclusions

The system can be considered usable as a standard microscope, and also for occasional remote consultations.
Itraconazole associated quadriparesis and edema: a case report
Authors: Karadi Rangaprasad L , Gow David , Kellett Mark , Denning David W , O'Driscoll Ronan B
Published: April 2011

Introduction

Itraconazole is an anti-fungal agent widely used to treat various forms of mycosis. It is particularly useful in allergic bronchopulmonary aspergillosis and severe asthma with fungal sensitization. Side effects are uncommon and usually mild. Mild neuropathy is noted to occur very rarely. We present an unusual and, to the best of our knowledge, as yet unreported case of severe neuropathy and peripheral edema due to itraconazole in the absence of a concomitant risk factor.

Case presentation

A 72-year-old Caucasian man was started on itraconazole following diagnosis of severe asthma with fungal sensitization. One month later he presented with severe bilateral ankle edema with an elevated serum itraconazole level. The itraconazole dose was reduced but his ankle edema persisted and he developed weakness of all four limbs. Itraconazole was completely stopped leading to improvement in his leg edema but he became bed bound due to weakness. He gradually improved with supportive care and neurorehabilitation. On review at six months, our patient was able to mobilize with the aid of two elbow crutches and power had returned to 5/5 in distal extremities and 4+/5 in proximal extremities. The diagnosis was established based on the classical presentation of drug-induced neuropathy and negative investigatory findings for any alternative diagnoses.

Conclusion

We report the case of a patient presenting with an unusual complication of severe neuropathy and peripheral edema due to itraconazole. Clinicians should be alert to this association when encountered with neuropathy and/or edema in an itraconazole therapy recipient.
Indeterminate and discrepant rapid HIV test results in couples' HIV testing and counselling centres in Africa
Authors: Boeras Debrah I , Luisi Nicole , Karita Etienne , McKinney Shila , Sharkey Tyronza , Keeling Michelle , Chomba Elwyn , Kraft Colleen , Wall Kristin , Bizimana Jean , Kilembe William , Tichacek Amanda , Caliendo Angela M , Hunter Eric , Allen Susan
Published: April 2011

Background

Many HIV voluntary testing and counselling centres in Africa use rapid antibody tests, in parallel or in sequence, to establish same‐day HIV status. The interpretation of indeterminate or discrepant results between different rapid tests on one sample poses a challenge. We investigated the use of an algorithm using three serial rapid HIV tests in cohabiting couples to resolve unclear serostatuses.

Methods

Heterosexual couples visited the Rwanda Zambia HIV Research Group testing centres in Kigali, Rwanda, and Lusaka, Zambia, to assess HIV infection status. Individuals with unclear HIV rapid antibody test results (indeterminate) or discrepant results were asked to return for repeat testing to resolve HIV status. If either partner of a couple tested positive or indeterminate with the screening test, both partners were tested with a confirmatory test. Individuals with indeterminate or discrepant results were further tested with a tie‐breaker and monthly retesting. HIV‐RNA viral load was determined when HIV status was not resolved by follow‐up rapid testing. Individuals were classified based on two of three initial tests as “Positive”, “Negative” or “Other”. Follow‐up testing and/or HIV‐RNA viral load testing determined them as “Infected”, “Uninfected” or “Unresolved”.

Results

Of 45,820 individuals tested as couples, 2.3% (4.1% of couples) had at least one discrepant or indeterminate rapid result. A total of 65% of those individuals had follow‐up testing and of those individuals initially classified as “Negative” by three initial rapid tests, less than 1% were resolved as “Infected”. In contrast, of those individuals with at least one discrepant or indeterminate result who were initially classified as “Positive”, only 46% were resolved as “Infected”, while the remainder was resolved as “Uninfected” (46%) or “Unresolved” (8%). A positive HIV serostatus of one of the partners was a strong predictor of infection in the other partner as 48% of individuals who resolved as “Infected” had an HIV‐infected spouse.

Conclusions

In more than 45,000 individuals counselled and tested as couples, only 5% of individuals with indeterminate or discrepant rapid HIV test results were HIV infected. This represented only 0.1% of all individuals tested. Thus, algorithms using screening, confirmatory and tie‐breaker rapid tests are reliable with two of three tests negative, but not when two of three tests are positive. False positive antibody tests may persist. HIV‐positive partner serostatus should prompt repeat testing.
Tuberculosis in cattle: the results of the four-area project
Authors: Griffin John M. , More Simon J. , Clegg Tracy A. , Collins John D. , O'Boyle Ian , Williams David H. , Kelly Gabrielle E. , Costello Eamon , Sleeman D. Paddy , O'Shea Finbarr , Duggan Murt , Murphy James , Lavin Desmond P.T.
Published: April 2011

The four-area project was undertaken to further assess the impact of badger removal on the control of tuberculosis in cattle herds in Ireland. It was conducted between 1997 and 2002 in matched removal and reference areas in four counties, namely Cork, Donegal, Kilkenny and Monaghan, representing a wide range of Irish farming environments. In the removal areas, a proactive programme of badger removal was conducted, on two or three occasions each year, whereas in the reference areas, badger removal was entirely reactive following severe outbreaks of tuberculosis amongst cattle. A detailed statistical analysis of this study has already been presented by Griffin et al. [13]; this paper presents further, mainly descriptive, findings from the study. In total, 2,360 badgers were captured in the removal areas of which 450 (19.5%) were considered positive for tuberculosis and 258 badgers were captured in the reference areas, with 57 (26.1%) positive for tuberculosis. The annual incidence of confirmed herd restrictions was lower in the removal area compared to the reference area in every year of the study period in each of the four counties. These empirical findings were consistent with the hazard ratios found by Griffin et al. [13]. Further, the effect of proactive badger removal on cattle tuberculosis in the four-area project and in the earlier east-Offaly project, as measured using the number of reactors per 1,000 cattle tested, were very similar, providing compelling evidence of the role of badgers in the epidemiology of tuberculosis in Irish cattle herds. The validity of the four-area project was discussed in detail. Efforts to minimise badger-to-cattle transmission in Ireland must be undertaken in association with the current comprehensive control programme, which has effectively minimised opportunities for cattle-to-cattle transmission.
Non-accidental injury in companion animals in the Republic of Ireland
Authors: McGuinness Kristina , Allen Mary , Jones Boyd R.
Published: April 2011

Non-accidental injury (NAI), animal abuse and "battered pet" syndrome are terms used to identify "the intentional harm of an animal". The terms include, but are not limited to, wilful neglect, inflicting injury, pain or distress, or malicious killing of an animal. Three categories of abuse are recognised: physical, sexual and neglect.

A postal survey was conducted to determine the extent to which NAI was recognised by veterinary surgeons in urban, semi-rural and rural veterinary practices in the Republic of Ireland. The questionnaire was sent to 600 veterinarians; completed submissions were received from 115 respondents (19.2%).

The occurrence of NAI was acknowledged by 106 (92.2%) of the respondents and cases had been seen by 50 (43.3%) of them, comprised of 36.2% of urban veterinary surgeons from rural towns and of 82% of urban practitioners. In 59% of cases the client indicated the injury was non-accidental; 39 (67.2%) of the 58 reported cases involved a single event. Signs that made veterinary surgeons suspicious of NAI included inconsistent history, untreated injuries, recurring injuries, meekness of the animal, suspicious behaviour of the owner and injuries consistent with abuse. The types of injuries observed included burns, lacerations, gunshot wounds, poisoning, injury to genitalia, bruising and fractures.

The findings of this study are comparable with those from other countries. Most but not all veterinary surgeons in Ireland recognise NAI and animal abuse is of significant concern in rural and urban communities as evidenced by this survey of practising veterinary surgeons.
Measurements of the acid-binding capacity of ingredients used in pig diets
Authors: Lawlor Peadar G. , Lynch P. Brendan , Caffrey Patrick J. , O'Reilly James J. , O'Connell M. Karen
Published: April 2011

Some feed ingredients bind more acid in the stomach than others and for this reason may be best omitted from pig starter foods if gastric acidity is to be promoted. The objective of this study was to measure the acid-binding capacity (ABC) of ingredients commonly used in pig starter foods. Ingredients were categorised as follows: (i) milk products (n = 6), (ii) cereals (n = 10), (iii) root and pulp products (n = 5), (iv) vegetable proteins (n = 11), (v) meat and fish meal (n = 2), (vi) medication (n = 3), (vii) amino acids (n = 4), (viii) minerals (n = 16), (ix) acid salts (n = 4), (x) acids (n = 10). A 0.5 g sample of food was suspended in 50 ml distilled de-ionised water with continuous stirring. This suspension was titrated with 0.1 mol/L HCl or 0.1 mol/L NaOH so that approximately 10 additions of titrant was required to reach pH 3.0. The pH readings after each addition were recorded following equilibration for three minutes. ABC was calculated as the amount of acid in milliequivalents (meq) required to lower the pH of 1 kg food to (a) pH 4.0 (ABC-4) and (b) pH 3.0 (ABC-3). Categories of food had significantly different (P < 0.01) ABC values. Mean ABC-4 and ABC-3 values of the ten categories were: (i) 623 (s.d. 367.0) and 936 (s.d. 460.2), (ii) 142 (s.d. 79.2) and 324 (s.d. 146.4), (iii) 368 (s.d. 65.3) and 804 (s.d. 126.7), (iv) 381 (s.d. 186.1) and 746 (s.d. 227.0), (v) 749 (s.d. 211.6) and 1508 (s.d. 360.8), (vi) 120 (s.d. 95.6) and 261 (s.d. 163.2), (vii) 177 (s.d. 60.7) and 1078 (s.d. 359.0), (viii) 5064 (s.d. 5525.1) and 7051 (s.d. 5911.6), (ix) 5057 (s.d. 1336.6) and 8945 (s.d. 2654.1) and (x) -5883 (s.d. 4220.5) and -2591 (s.d. 2245.4) meq HCl per kg, respectively. Within category, ABC-3 and ABC- 4 values were highly correlated: R2 values of 0.80 and greater for food categories i, iv, v, vi, vii and viii. The correlation between predicted and observed ABC values of 34 mixed diets was 0.83 for ABC-4 and 0.71 for ABC-3. It was concluded that complete diets with low ABC values may be formulated through careful selection of ingredients. The final pH to which ABC is measured should matter little as ABC-3 and ABC-4 are highly correlated.
A folate receptor beta-specific human monoclonal antibody recognizes activated macrophage of rheumatoid patients and mediates antibody-dependent cell-mediated cytotoxicity
Authors: Feng Yang , Shen Jiayin , Streaker Emily D , Lockwood Michael , Zhu Zhongyu , Low Philip S , Dimitrov Dimiter S
Published: April 2011

Introduction

Folate receptor beta (FRβ) is only detectable in placenta and limited to some hematopoietic cells of myeloid lineage in healthy people. Studies have indicated that FRβ is over-expressed in activated macrophages in autoimmune diseases and some cancer cells. In this study we aimed to develop an FRβ-specific human monoclonal antibody (mAb) that could be used as a therapeutic agent to treat rheumatoid arthritis and other autoimmune diseases, as well as FRβ positive cancers.

Methods

Functional recombinant FRβ protein was produced in insect cells and used as antigen to isolate a mAb, m909, from a human naïve Fab phage display library. Binding of Fab and IgG1 m909 to FRβ was measured by ELISA, surface plasmon resonance, immune fluorescence staining, and flow cytometry. Antibody-dependent cell-mediated cytotoxicity (ADCC) was evaluated with FRβ positive CHO cells as target cells and isolated peripheral blood monocytes as effector cells in an in vitro assay.

Results

Fab m909 bound with relatively high affinity (equilibrium dissociation constant 57 nM) to FRβ. The IgG1 m909 showed much higher (femtomolar) avidity as measured by ELISA, and it bound to FRβ positive cells in a dose-dependent manner, but not to parental FRβ negative cells. m909 did not compete with folate for the binding to FRβ on cells. m909 was not only able to select FRβ positive, activated macrophages from synovial fluid cells of arthritis patients as efficiently as folate, but also able to mediate ADCC in FRβ positive cells.

Conclusions

Unlike folate-drug conjugates, m909 selectively binds to FRβ, does not recognize FRα, and has at least one effector function. m909 alone has potential to eliminate FRβ positive cells. Because m909 does not compete with folate for receptor binding, it can be used with folate-drug conjugates in a combination therapy. m909 can also be a valuable research reagent.
Serum concentrations of insulin-like growth factor-I, insulin-like growth factor binding protein-3 and cardiovascular risk factors in adolescents
Authors: Kong Alice P S , Choi Kai-Chow , Wong Gary W K , Ko Gary T C , Ho Chung-Shun , Chan Michael H M , Ozaki Risa , Ma Ronald C W , Lau Joseph T F , Chan Juliana C N
Published: April 2011

Background

The risk association between the insulin like growth factor-I (IGF-I) system and cardiovascular risk is inconclusive in adults and under-explored in adolescents. We aimed to investigate the associations between serum concentrations of IGF-I and IGF binding protein-3 (IGFBP-3) and cardiovascular risk factors in adolescents.

Methods

This was a cross-sectional, population-based, observational study in a school setting with 2102 Hong Kong Chinese adolescents aged 12–19 years. Serum IGF-I and IGFBP-3 concentrations were measured by chemiluminescence immunoassays. Anthropometric indices and traditional cardiovascular risk factors were assessed.

Results

After excluding participants with abnormal thyroid and liver test results, 765 boys and 877 girls, mean (±SD) age of 15.3 (±2.0) and 15.7 (±2.0) years, respectively, were included in the analysis. Multivariable regression analyses revealed that both IGF-I and IGFBP-3 concentrations were independently associated with waist circumference, fasting insulin and haemoglobin concentrations in boys (all P < 0.05), systolic blood pressure, serum creatinine, fasting insulin and haemoglobin concentrations in girls (all P < 0.05). In girls, IGF-I was also associated with C-reactive protein concentration ( P < 0.001) and IGFBP-3 was associated with fasting triglyceride concentration ( P < 0.001). Compared with adolescents with the lowest tertile, the top tertile of both IGF-I and IGFBP-3 concentrations were associated with increased odds of having overweight/obesity, top tertiles of insulin and haemoglobin in both boys and girls ( P for trend, all <0.05).

Conclusions

The associations between serum IGF-I, IGFBP-3, obesity, cardiovascular risk factors, insulin and haemoglobin suggest that dysregulation of the IGF system may play a linking role for the clustering of cardiovascular risk factors.
Update on investigating hypoglycaemia in childhood
Authors: Lang Timothy F
Published: April 2011

Identification and investigation of hypoglycaemia in childhood remains an important clinical emergency. Rapid recognition and appropriate management of this clinical state continues to be important in order to prevent neurological damage or even death. The purpose of this review is to provide an update on the advances made in this area since the review by Bonham in this journal in 1993. Advances in molecular science and diagnostic techniques have assisted in understanding the mechanisms involved in the homeostasis of glucose metabolism at rest and when stressed. New disorders causing hypoglycaemia are described using the classification based upon aetiologies, which was used in Bonham's original paper. The development and use of guidelines and pre-assembled packs for investigating hypoglycaemia is also discussed.
Isoangustone A suppresses mesangial fibrosis and inflammation in human renal mesangial cells
Authors: Li Jing , Lim Soon Sung , Lee Eun-Sook , Gong Ju-Hyun , Shin Daekeun , Kang Il-Jun , Kang Young-Hee
Published: April 2011

Development of diabetic nephropathy with fibrosis is associated with hypereglycemia-linked inflammation. Increased levels of proinflammatory factors have been found in diabetic patients with nephropathy. The present study was to test the hypothesis that isoangustone A, a novel compound present in licorice, can inhibit renal fibrosis and inflammation inflamed by high glucose (HG) in human mesangial cells through disturbing transforming growth factor β (TGF- β) and nuclear facor κB (NF- κB) pathways. Serum-starved mesangial cells were cultured in 33 mmol/L glucose media. Cells were treated with 1–20 μmol/L isoangustone A isolated from Glycyrrhiza uralensis licorice for three days. Exposure of cells to HG elevated connective tissue growth factor and collagen production, which was dose-dependently reversed by isoangustone A. Isoangustone A boosted HG-plummeted membrane type matrix metalloproteinase (MMP)-1 expression and diminished HG-elevated tissue inhibitor of MMP-2 expression. HG activated mesangial TGF- β1-SMAD-responsive signaling, which was repealed by ≥10 μmol/L isoangustone A. Furthermore, HG upregulated intracellular cell adhesion molecule-1 (ICAM-1) level and monocyte chemoattractant protein-1 (MCP-1) mRNA expression, and such increases were dose-dependently suppressed by isoangustone A most likely through hampering TGF- β signaling pathways. Blockade of NF- κB signaling appeared to be responsible for attenuating HG-triggered induction of ICAM-1 and MCP-1. Our findings provide the first evidence that isoangustone A dampens mesangial sclerosis associated with inflammation in response to HG through hindering TGF- β and NF- κB signaling.
Perinatal exposure to low-dose DE-71 increases serum thyroid hormones and gonadal osteopontin gene expression
Authors: Blake Charles A , McCoy George L , Hui Yvonne Y , LaVoie Holly A
Published: April 2011

Polybrominated diphenyl ethers (PBDEs) are flame retardants that have been widely used in manufacturing. They are major household and environmental contaminants that bioaccumulate. Humans are exposed primarily through dust inhalation and dietary ingestion of animal products. In animal studies, high doses of penta-brominated diphenyl ethers (penta-BDEs) in the mg/kg body weight (BW) range negatively impact brain development, behavior, memory, circulating thyroid hormone concentrations, the reproductive system and bone development. We investigated the effects of ingestion of a relatively low dose of the penta-BDE mixture DE-71 by pregnant and lactating rats on reproductive and thyroid parameters of the F1 offspring. F0 mothers received 60 μg/kg BW of DE-71 or vehicle daily by gavage from Day 1.5 of pregnancy through lactation (except the day of parturition). F1 pups were sacrificed at 21 d of age or outbred at approximately 80 d of age. Bred F1 females were sacrificed at Day 14.5 of pregnancy or at five months of age. Bred F1 males were sacrificed at five months of age. DE-71 treatment of the mothers affected the F1 females as evidenced by lower body weights at 80 d and five months of age, elevated serum T3 and T4 concentrations at Day 14.5 of pregnancy and increased thyroid gland weight and ovarian osteopontin mRNA at five months of age. Perinatal DE-71 exposure also increased testicular osteopontin mRNA in 21-day-old F1 males. Utilizing a granulosa cell in vitro model, we demonstrated that DE-71 activated the rat osteopontin gene promoter. Our results are the first to demonstrate that PBDEs increase rodent circulating T3 and T4 concentrations and gonadal osteopontin mRNA, and activate the osteopontin gene promoter. These changes may have clinical implications as others have shown associations between human exposure to PBDEs and subclinical hyperthyroidism, and overexpression of ovarian osteopontin has been associated with ovarian cancer.
Improved engraftment with minimal graft-versus-host disease after major histocompatibility complex-mismatched cord blood transplantation with photochemically treated donor lymphocytes
Authors: Kanathezhath Bindu , Mizokami Myra , Stanislaus Sharleny , Hounshell Catherine , Neumayr Lynne , Guo Hua , Hearst John E , Walters Mark C , Kuypers Frans A
Published: April 2011

There is a significant risk of severe graft-versus-host disease (GVHD) and graft failure after unrelated umbilical cord blood transplantation (CBT) if donor–recipient pairs are mismatched at major histocompatibility complex (MHC) loci. To mitigate these risks after MHC-mismatched CBT, we infused psoralen-treated, photochemically inactivated, mature donor T-lymphocytes with MHC (H2-haplotype) mismatched murine donor fetal near-term peripheral blood (FNPB) cells after sublethal irradiation. We analyzed the rates of donor engraftment, GVHD and long-term survival in H2 haplotype disparate (C57BL/6 [H-2b/Thy1.1] → AKR [H-2k/Thy1.2]) recipient mice. We observed inconsistent donor engraftment after transplantation with cord blood alone, but superior engraftment and long-term survival after FNPB transplantation supplemented with psoralen-treated donor T-lymphocytes. Additionally, there was fatal GVHD after FNPB co-infusion with untreated donor T-lymphocytes, but minimal GVHD after FNPB supplemented with psoralen-treated donor T-lymphocytes transplantation. Donor MHChigh/c-Kit+/lineage−/CD34−stem cells were noted in the recipient bone marrow compartment following co-infusion of photochemically inactivated T-cells with FNPB. Despite the non-myeloablative preparation before FNPB infusion, complete hematological recovery was delayed until 50–60 d after transplantation. We observed that co-transplantation of psoralen-treated donor T-lymphocytes with FNPB facilitated durable engraftment of donor hematopoietic stem cells in the marrow and splenic compartments with complete but delayed recovery of all hematopoietic lineages. This CBT model establishes the possibility of ensuring donor engraftment across a MHC barrier without severe GVHD.
Preclinical evaluation of gene delivery methods for the treatment of loco-regional disease in breast cancer
Authors: Rajendran Simon , O'Hanlon Deirdre , Morrissey David , O'Donovan Tracey , O'Sullivan Gerald C , Tangney Mark
Published: April 2011

Preclinical results with various gene therapy strategies indicate significant potential for new cancer treatments. However, many therapeutics fail at clinical trial, often due to differences in tissue physiology between animal models and humans, and tumor phenotype variation. Clinical data relevant to treatment strategies may be generated prior to clinical trial through experimentation using intact patient tissue ex vivo. We developed a novel tumor slice model culture system that is universally applicable to gene delivery methods, using a realtime luminescence detection method to assess gene delivery. Methods investigated include viruses (adenovirus [Ad] and adeno-associated virus), lipofection, ultrasound (US), electroporation and naked DNA. Viability and tumor populations within the slices were well maintained for seven days, and gene delivery was qualitatively and quantitatively examinable for all vectors. Ad was the most efficient gene delivery vector with transduction efficiency >50%. US proved the optimal non-viral gene delivery method in human tumor slices. The nature of the ex vivo culture system permitted examination of specific elements. Parameters shown to diminish Ad gene delivery included blood, regions of low viability and secondary disease. US gene delivery was significantly reduced by blood and skin, while tissue hyperthermia improved gene delivery. US achieved improved efficacy for secondary disease. The ex vivo model was also suitable for examination of tissue-specific effects on vector expression, with Ad expression mediated by the CXCR4 promoter shown to provide a tumor selective advantage over the ubiquitously active cytomegalovirus promoter. In conclusion, this is the first study incorporating patient tissue models in comparing gene delivery from various vectors, providing knowledge on cell-type specificity and examining the crucial biological factors determining successful gene delivery. The results highlight the importance of in-depth preclinical assessment of novel therapeutics and may serve as a platform for further testing of current, novel gene delivery approaches.
Reference values of biochemical and hematological parameters for Guizhou minipigs
Authors: Chen Younan , Qin Shengfang , Ding Yang , Li Shengfu , Yang Guang , Zhang Jie , Li Youping , Cheng Jingqiu , Lu Yanrong
Published: April 2011

The pig is not only an economically important livestock animal but is also a valuable model animal for biomedical research and xenotransplantation. Reference values for clinical biochemical and hematological parameters are required for accurate data interpretation while using a pig model. In this study, whole blood samples were collected from 54 healthy Chinese Guizhou minipigs. We analyzed routine biochemical and hematological parameters and special coagulation parameters, including thrombelastography and coagulation factor activities, and have presented the baseline values of these parameters. These data provide valuable information for investigators using minipigs as animal models in biomedical studies and useful physiological data for veterinarians and livestock producers. We also compared all the results for the minipigs with the corresponding data from healthy humans. The bilirubin, uric acid and cholesterol levels of minipigs were significantly lower than those of humans (14%, 0.086% and 48% of human levels, respectively), whereas the serum enzyme levels were much higher than those in humans (e.g. the hydroxybutyrate dehydrogenase and creatine kinase levels of the minipigs were 19- and 8.4-fold higher than the human reference values). The red blood cell counts, platelet counts and white blood cell counts of the minipigs were significantly higher than those of the humans. The coagulation activities of factor VII and factor X were higher in minipigs than in humans. The significant differences observed between minipigs and humans for many of these parameters suggest substantial interspecies disparities in organs and tissues. These differences merit greater attention in biomedical research involving minipigs, particularly in the area of pig-to-human transplantation.
STAT1 promotes radioresistance of CD44+/CD24−/low cells in breast cancer
Authors: Zhan Jun-fang , Chen Long-hua , Yuan Ya-wei , Xie Guo-zhu , Sun Ai-min , Liu Ying , Chen Zhi-xian
Published: April 2011

Breast cancer-initiating cells are a relatively radioresistant subpopulation of breast cancer cells. However, the mechanism of this radioresistance is still unclear. This study aimed to investigate the effect of radiation on the levels of signal transducer and activator of transcription 1 (STAT1) in mammospheres of cancer-initiating cells and monolayer cultures of MCF-7 cells. We isolated CD44+/CD24−/low cancer-initiating cells from MCF-7 cells and propagated them as mammospheres. Next we used realtime quantitative reverse-transcriptase polymerase chain reaction to examine the mRNA level of STAT1 in mammospheres of breast cancer-initiating cells and monolayer cultures of MCF-7 cells. The apoptosis rate and surviving fraction using clonogenic assays was observed after treating the cells with a STAT1 inhibitor. After irradiation, the STAT1 level in the mammospheres was higher than that in the monolayer cultures. STAT1 inhibitor treatment did not cause significant changes in the apoptosis rate and surviving fraction in the MCF-7 monolayer cultures. However, the inhibitor treatment caused significant differences in the apoptosis rate and surviving fraction in mammospheres. Our study provides the first evidence that STAT1 signaling contributes to radioresistance in breast cancer-initiating cells and reveals STAT1 as a promising target to reduce radioresistance and enhance the efficacy of radiotherapy for breast cancer.
Platelet activating factor: the good and the bad in the ischemic/reperfused heart
Authors: Penna Claudia , Bassino Eleonora , Alloatti Giuseppe
Published: April 2011

The present review is focused on the dual role played by platelet-activating factor (PAF) in ischemia and reperfusion (I/R) injury of the heart. Although the involvement of PAF in the pathogenesis of myocardial reperfusion injury is well established, in the last few years it has emerged that very low concentrations of PAF exert cardioprotective effects, comparable to that afforded by ischemic preconditioning (IP). PAF is a potent phosphoglyceride involved in different pathophysiological conditions affecting the cardiovascular system, including the development of myocardial I/R injury. PAF is released from the I/R myocardium in concentrations (1–10 nmol/L) high enough to negatively modulate coronary circulation as well as electrical and contractile activities. PAF may act either directly, via generation of secondary mediators, or through the activation of inflammatory cells like platelets and polymorphonuclear neutrophils, which exacerbate postischemic myocardial injury. The effects of PAF are mediated through specific receptors (PAFRs) that belong to the superfamily of G protein-coupled receptors. Since cardiomyocytes not only produce PAF but also possess PAFRs, it is likely that PAF acts as an autocrine/paracrine mediator. Although the negative effects exerted by high concentrations of PAF are well established, several recent findings from our and other laboratories have demonstrated that very low concentrations (pmol/L) of PAF infused before ischemia induce cardioprotective effects similar to those afforded by IP, and that endogenous PAF production participates in the induction of IP itself. The IP-like action exerted by low concentrations of PAF is due to the activation/phosphorylation of kinases included in the reperfusion injury salvage kinase (RISK) pathway, such as protein kinase C, Akt/PkB and nitric oxide synthase. Together with the activation of mitochondrial KATPchannels, these events may allow prevention of mitochondrial permeability transition pores opening at reperfusion. Moreover, the nitric oxide-dependent S-nitrosylation of L-type Ca2+channels induced by PAF reduces intracellular Ca2+overload.
Selection and characterization of DNA aptamers against PrPSc
Authors: Wang Ping , Hatcher Kristen L , Bartz Jason C , Chen Shu G , Skinner Pamela , Richt Juergen , Liu Hong , Sreevatsan Srinand
Published: April 2011

Transmissible spongiform encephalopathies (TSEs) are a group of zoonotic and fatal neurodegenerative disorders that affect humans and animals. The pathogenesis of TSEs involves a conformational conversion of the cellular prion protein (PrP) into abnormal isoforms. Currently, cellular and pathological forms of PrP are differentiated using specific antibody-based analyses that are resource intensive and not applicable to all species and strains. Thus, there is an urgent need for sensitive and efficient assays that can detect pathological forms of PrP. Using systematic evolution of ligands by exponential enrichment, we developed DNA aptamers that can differentiate normal and abnormal PrP isoforms. These aptamers represent the first reagents that can identify pathological isoforms of PrP across multiple host species. Second, they are able to distinguish different strains of prions. Third, they can be used to detect prions in peripheral blood cells, which are otherwise undetectable using conventional antibody-based detection methods. Thus, DNA aptamers offer promise for the development of presymptomatic screens of tissue, blood and other body fluids for prion contamination.
Analysis of plasma membrane Ca2+-ATPase gene expression during epileptogenesis employing single hippocampal CA1 neurons
Authors: Bravo-Martínez Jorge , Delgado-Coello Blanca , García David E , Mas-Oliva Jaime
Published: April 2011

Disruption of calcium homeostasis in epileptic cells is characterized by both short- and long-term perturbations of Ca2+ buffering systems. Along with the Na+/Ca2+ exchanger, the plasma membrane Ca2+-ATPase (PMCA) plays an important role in excitable cells. The involvement of PMCAs in epileptogenesis has primarily been studied in brief intervals after various stimuli; however, the specific contribution of this molecule to epileptogenesis is not yet fully understood. Our aim has been to investigate whether PMCA expression in the chronic stages of epilepsy is altered. Through an interdisciplinary approach, involving whole-cell recordings and real-time reverse transcriptase-polymerase chain reaction, we have shown that epileptic neurons in our preparation consistently show changes in electrical properties during the period of chronic epilepsy. These changes included increased spike frequency, altered resting membrane potential and changes in passive membrane properties. Following these observations, which indicate an altered excitability in the epileptic cells studied, PMCA mRNA transcripts were studied. It was found that while PMCA1 transcripts are significantly increased one month following the pilocarpine epileptogenic stimulus, PMCA3, an isoform important in excitable tissues, was significantly, decreased. These findings suggest that, in the long-term, a slow PMCA ( PMCA1) plays a role in the reestablishment of a new calcium homeostasis attained by epileptic cells. Overall, this phenomenon points out the fact that in seizure disorders, changes that take place in the balance of the different molecules and their isoforms in charge of maintaining neuronal calcium homeostasis, are fundamental in the survival of affected cells.
Role of AMP-activated protein kinase in the control of hepatocyte priming and proliferation during liver regeneration
Authors: Varela-Rey Marta , Beraza Naiara , Lu Shelly C , Mato José M , Martínez-Chantar María L
Published: April 2011

The enzyme AMP-activated protein kinase (AMPK) is the main energy sensor in cells and is responsible for controlling the balance of anabolic/catabolic processes under metabolic stress conditions. This metabolic control exerted by AMPK is critical for energy-demanding situations, such as liver regeneration. Immediately after partial hepatectomy (PH), the liver undergoes the priming phase, mediated by the proinflammatory cytokines tumor necrosis factor (TNF) and interleukin-6, which promote responsiveness of hepatocytes to growth factors, such as hepatocyte growth factor (HGF) and epidermal growth factor, which lead to proliferation. In addition to its metabolic function, AMPK is likely to be a key mediator in both hepatocyte priming and the proliferative phases, induced by TNF- α and HGF, respectively. TNF- α-induced AMPK activation has been shown to be necessary for nuclear factor κappa B (NF- κB)-induced inducible nitric oxide synthase expression and for blocking TNF- α-induced apoptosis. On the other hand, HGF-induced LKB1/AMPK activation has been found to play a critical role in controlling Hu antigen R cytosolic localization and endothelial nitric oxide synthase activation, and consequently Cyclin D1 and Cyclin A expressions, and nitric oxide generation, respectively. During PH, levels of S-adenosylmethionine (SAMe), the principal methyl donor in the liver, have to decrease to allow liver proliferation. Our studies also show that SAMe inhibits hepatocyte proliferation by controlling the hepatocyte's responsiveness to mitogenic signals such as HGF through the inhibition of AMPK activity. In summary, these data highlight the essential role of AMPK in controlling the balance between hepatocyte metabolic adaptations, cell cycle progression and apoptosis during liver regeneration.
Pyridoxal 5′ phosphate protects islets against streptozotocin-induced beta-cell dysfunction – in vitro and in vivo
Authors: Kiran Sashi G , Dorisetty Raj K , Umrani Malathi R , Boindala Sesikeran , Bhonde Ramesh R , Chalsani Maniprabha , Singh Himadri , Venkatesan Vijayalakshmi
Published: April 2011

Administration of pyridoxal 5′ phosphate (PLP) has demonstrated beneficial effects in the management of diabetes, albeit the mechanism(s) are not clearly understood. The present study addressed the islet-cell function(s) in streptozotocin (STZ)-induced diabetic mice both in vitro and in vivo. Primary islet cells primed with or without PLP (5 mmol/L) were treated with STZ (2 mmol/L) and were measured for cell viability, insulin secretion, free radicals and mRNA of Insulin and Pdx1. The specificity of PLP's response on insulin secretion was assessed with amino oxy acetic acid (AOAA)-PLP inhibitor. In vivo, the STZ (200 mg/kg b.w)-treated diabetic mice received 10 mmol/L PLP intraperitoneally a day before (PLP + STZ) or after (STZ + PLP) with three more doses once every 48 h. On 7, 14 and 21 d of STZ treatment, physiological parameters, islet morphology, insulin:glucagon, insulin:HSP104, and mRNA of Insulin, Glut2, Pdx1 and Reg1 were determined. In vitro, PLP protected islets against STZ-induced changes in viability, insulin secretion, prevented increase in free radical levels and normalized mRNA of Insulin and Pdx1. Further, AOAA inhibited PLP-induced insulin secretion in islets. In vivo, PLP treatment normalized STZ-induced changes in physiological parameters, circulating levels of PLP and insulin. Also, islet morphology, insulin:glucagon, insulin:HSP104 and mRNA levels of Insulin, Pdx1 and Glut2 were restored by 21 d. Although PLP treatment (pre- and post-STZ) prevented development of frank diabetes, STZ + PLP mice showed transient hyperglycemia, and increased mRNA for Reg1. The data suggest the cytoprotective vis-à-vis insulinotrophic effects of PLP against STZ-induced beta-cell dysfunction and underline its prophylactic use in the management of diabetes.
Single dose streptozotocin-induced diabetes: considerations for study design in islet transplantation models
Authors: Deeds M C , Anderson J M , Armstrong A S , Gastineau D A , Hiddinga H J , Jahangir A , Eberhardt N L , Kudva Y C
Published: April 2011

Streptozotocin (STZ)-induced diabetes mellitus (DM) offers a very cost-effective and expeditious technique that can be used in most strains of rodents, opening the field of DM research to an array of genotypic and phenotypic options that would otherwise be inaccessible. Despite widespread use of STZ in small animal models, the data available concerning drug preparation, dosing and administration, time to onset and severity of DM, and any resulting moribundity and mortality are often limited and inconsistent. Because of this, investigators inexperienced with STZ-induced diabetes may find it difficult to precisely design new studies with this potentially toxic chemical and account for the severity of DM it is capable of inducing. Until a better option becomes available, attempts need to be made to address shortcomings with current STZ-induced DM models. In this paper we review the literature and provide data from our pancreatic islet transplantation experiments using single high-dose STZ-induced DM in NCr athymic nude mice with hopes of providing clarification for study design, suggesting refinements to the process, and developing a more humane process of chemical diabetes induction.
How do rats respond to playing radio in the animal facility?
Authors: Krohn Thomas C , Salling Bo , Hansen Axel Kornerup
Published: April 2011

In the animal facility, a range of different sounds are present. On the one hand, rats and humans will regard sound and noise differently even within the audible range, but on the other hand mice and rats being very adaptable to the environment may adapt to living in a noisy facility with e.g. a radio playing. It was the aim of the present study to investigate whether two different strains of rats had different preferences for different kinds of sound patterns, including radio, and to get an indication of whether they are able to distinguish between different sound patterns. The present preference study revealed that rats were able to distinguish between different sound patterns. They showed a clear preference for silence to anything else, which may be taken as an indication that they feel disturbed by the sound from the speaker.