bluuebunny/crossref_metadata_2025 huggingface dataset:
Improvement in Cheyne-Stokes respiration following cardiac resynchronisation therapy
Authors: Gabor J. Y. , Newman D. A. , Barnard-Roberts V. , Korley V. , Mangat I. , Dorian P. , Hanly P. J.
Published: July 2005

The effect of standard cardiac resynchronisation therapy (CRT) on the severity of Cheyne-Stokes respiration (CSR) in patients with congestive heart failure was studied. It was hypothesised that CRT, through its known beneficial effects on cardiac function, would stabilise the control of breathing and reduce CSR.

Twenty-eight patients who were eligible for CRT and receiving optimised medical treatment for congestive heart failure were referred for overnight polysomnography, including monitoring of thoracic and abdominal movements to identify CSR and obstructive sleep apnoea events. Patients underwent repeat polysomnography after 6 months of CRT to re-evaluate sleep quality and sleep-disordered breathing.

Twelve of the 28 patients had significant CSR (43%); 10 patients had a successful implantation and underwent repeat polysomnography a mean±sd27±7 weeks after continuous biventricular pacing. Six of the 10 patients experienced a significant decrease in CSR severity following CRT, associated with correction of congestive heart failure-related hyperventilation and hypocapnia. Circulation time, oxygen saturation, frequency of obstructive apnoeas and sleep quality did not change.

In conclusion, cardiac resynchronisation therapy is associated with a reduction in Cheyne-Stokes respiration, which may contribute to improved clinical outcome in patients treated with cardiac resynchronisation therapy.
Early gastrostomy reduces the rate of ventilator-associated pneumonia in stroke or head injury patients
Authors: Kostadima E. , Kaditis A. G. , Alexopoulos E. I. , Zakynthinos E. , Sfyras D.
Published: July 2005

Presence of a nasogastric tube is a risk factor for the development of ventilator-associated pneumonia (VAP). Alternatively, gastrostomy can be used for administration of enteral feedings.

To determine whether early performance of gastrostomy affects frequency of VAP, a randomised, controlled study was carried out in patients mechanically ventilated for stroke or head injury. In the gastrostomy group, patients underwent the procedure within 24 h of intubation. A nasogastric tube was inserted in controls. Individual subjects were studied for 3 weeks.

In total, 20 subjects (mean age 48±15.2 yrs) were allocated to the gastrostomy group, and 21 to the control group (46.6±15.4 yrs). Of these groups, two (10%) and eight (38.1%) developed VAP, respectively. Four patients with gastrostomy and three controls did not complete the study (due to weaning from ventilatory support or death). After excluding these subjects, difference in VAP frequency persisted: two out of 16 subjects with gastrostomy had VAP (12.5%)versuseight out of 18 controls (44.4%). There were no differences in duration of hospitalisation or mortality between the two groups.

In conclusion, in patients mechanically ventilated for stroke or head injury early gastrostomy is associated with a lower frequency of ventilator-associated pneumonia compared with a nasogastric tube.
The dream of a vaccine against tuberculosis; new vaccines improving or replacing BCG?
Authors: Martín C.
Published: July 2005

In the last 10 yrs, work with experimental laboratory models has developed many new vaccine candidates against tuberculosis (TB). They include subunit vaccines, modified bacilli Calmette-Guérin (BCG) and attenuatedMycobacterium tuberculosis. Phase I clinical trials of new TB vaccine candidates have begun for the first time after 80 yrs of use of BCG. Many of these new trials involve recombinant BCG or improve BCG immunity by boosting with vaccines consisting of subunits or attenuated vaccinia virus expressing TB antigens.

However, effective vaccination against TB presents diverse and complex challenges. For example, TB infection can become reactivated years later and infection does not guarantee resistance to a subsequent second infection. A truly effective TB vaccine may, therefore, have to elicit an immune response that is greater than that induced by natural infection. In addition, various different populations have to be protected including those vaccinated with BCG and those infected withM. tuberculosisor HIV.

The goal is a new generation of vaccines effective against respiratory forms of tuberculosis. As a first step, good candidate vaccines able to boost bacille Calmette-Guérin and thereby improve protection could be a reality in the near future. Tuberculosis vaccine candidates, able to replace the currently used bacille Calmette-Guérin and/or make the eradication of tuberculosis feasible, can only be expected in the long-term, and safe live vaccines could be promising candidates.
Modulation of airway inflammation and resistance in mice by a nicotinic receptor agonist
Authors: Blanchet M-R. , Israël-Assayag E. , Cormier Y.
Published: July 2005

Nicotinic agonists, including 1,1-dimethyl-4-phenylpiperazinium (DMPP), have anti-inflammatory properties and in some instances smooth muscle relaxing effects. Since inflammation and airway smooth muscle contraction are two major components of asthma, the present authors investigated the effects of DMPP on airway inflammation and airway resistance in a mouse model of asthma.

Mice were sensitised and challenged with ovalbumin (OVA) and treated either intraperitoneally or intranasally with DMPP. The effect of DMPP was tested on airway inflammation, airway resistance and on the increase of intracellular calcium in bronchial smooth muscle cells.

DMPP given either during sensitisation, OVA challenges or throughout the protocol prevented lung inflammation and decreased the serum level of OVA specific immunoglobulin E. DMPP administration reduced the number of total cells, lymphocytes and eosinophils in the bronchoalveolar lavage (BAL) fluid. Intranasal DMPP administration was as effective as dexamethasone (DEXA) in reducing total cell count and eosinophil counts in BAL fluid. DMPP, but not DEXA, reduced tissue inflammation. Intranasal DMPP, given 10 min before the test, reduced airway responsiveness to metacholine. DMPP also reduced the increase in intracellular calcium in response to bradykinin.

In conclusion, these results show that 1,1-dimethyl-4-phenylpiperazinium reduces lung inflammation and prevents airway hyperresponsiveness in the mouse model of asthma.
The use of iloprost in early pregnancy in patients with pulmonary arterial hypertension
Authors: Elliot C. A. , Stewart P. , Webster V. J. , Mills G. H. , Hutchinson S. P. , Howarth E. S. , Bu'Lock F. A. , Lawson R. A. , Armstrong I. J. , Kiely D. G.
Published: July 2005

In patients with pulmonary hypertension, pregnancy is associated with a high risk of maternal death. Such patients are counselled to avoid pregnancy, or if it occurs, are offered early interruption. Some patients, however, decide to continue with their pregnancy and others may present with symptoms for the first time whilst pregnant. Pulmonary vasodilator therapy provides a treatment option for these high-risk patients.

The present study describes three patients with pulmonary arterial hypertension of various aetiologies who were treated with the prostacyclin analogue iloprost during pregnancy, and the post-partum period.

Nebulised iloprost commenced as early as 8 weeks of gestation and patients were admitted to hospital between 24–36 weeks of gestation. All pregnancies were completed with a duration of between 25–36 weeks and all deliveries were by caesarean section under local anaesthetic. All patients delivered children free from congenital abnormalities, and there was no post-partum maternal or infant mortality.

In conclusion, although pregnancy is strongly advised against in those with pulmonary hypertension, the current authors have achieved a successful outcome for mother and foetus with a multidisciplinary approach and targeted pulmonary vascular therapy.
Pulmonary manifestations of multicentric Castleman's disease in HIV infection: a clinical, biological and radiological study
Authors: Guihot A. , Couderc L-J. , Agbalika F. , Galicier L. , Bossi P. , Rivaud E. , Scherrer A. , Zucman D. , Katlama C. , Oksenhendler E.
Published: July 2005

The aim of the present study was to report clinical, radiological and bronchoalveolar lavage (BAL) findings in patients with pulmonary manifestations of HIV-associated multicentric Castleman's disease (MCD).

This was a retrospective study of 12 patients with histologically proven MCD.

Clinical manifestations were as follows: dyspnoea (nine out of 12 cases), cough (n = 10), bilateral crackles (n = 10), together with high fever, malaise, peripheral lymphadenopathy (n = 12), and hepatosplenomegaly (n = 10). Two patients developed acute respiratory distress syndrome. Chest radiographs and computed tomography scans showed reticular (n = 7) and/or nodular (n = 7) interstitial patterns, with mediastinal lymphadenopathy (n = 9), and bilateral pleural effusion (n = 3). Fibreoptic endoscopy was normal in all cases. BAL analysis showed hypercellularity (n = 6) and/or lymphocytosis (n = 6), and human herpesvirus-8 DNA was detected in two out of two cases. Specific stains and cultures for pathogens were negative. All patients received etoposide and/or vinblastine, and improved after 2–4 days. Relapses were frequent (50 attacks in 12 patients). Six patients developed a non-Hodgkin's lymphoma, and five died.

In conclusion, the pulmonary manifestation of HIV-related multicentric Castleman's disease is an acute reticulo-nodular interstitial pneumonitis, associated with severe systemic symptoms and peripheral lymphadenopathy. In bronchoalveolar lavage fluid, cellularity is not specific and human herpesvirus-8 DNA is detected. The clinical course is specific due to a rapid onset and regression, frequent relapses and a high occurrence of non-Hodgkin's lymphoma.
Effect of ventilation strategy and surfactant on inflammation in experimental pneumonia
Authors: van Kaam A. H. L. C. , Lutter R. , Lachmann R. A. , Haitsma J. J. , Herting E. , Snoek M. , De Jaegere A. , Kok J. H. , Lachmann B.
Published: July 2005

This study explored, the inflammatory response during experimental pneumonia in surfactant-depleted animals as a function of ventilation strategies and surfactant treatment.

Following intratracheal instillation of Group B streptococci (GBS), surfactant-depleted piglets were treated with conventional (positive-end expiratory pressure (PEEP) of 5 cmH2O, tidal volume 7 mL·kg−1) or open lung ventilation. During the latter, collapsed alveoli were recruited by applying high peak inspiratory pressures for a short period of time, combined with high levels of PEEP and the smallest possible pressure amplitude. Subgroups in both ventilation arms also received exogenous surfactant. Conventionally ventilated healthy animals receiving GBS and surfactant-depleted animals receiving saline served as controls.

In contrast with both control groups, surfactant-depleted animals challenged with GBS and conventional ventilation showed high levels of interleukin (IL)-8, tumour necrosis factor (TNF)-α and myeloperoxidase in bronchoalveolar lavage fluid after 5 h of ventilation. Open lung ventilation attenuated this inflammatory response, but exogenous surfactant did not. Systemic dissemination of the inflammatory response was minimal, as indicated by low serum levels of IL-8 and TNF-α.

In conclusion, the current study indicates that the ventilation strategy, but not exogenous surfactant, is an important modulator of the inflammation during Group B streptococci pneumonia in mechanically ventilated surfactant-depleted animals.
Overexpression of cathepsin f, matrix metalloproteinases 11 and 12 in cervical cancer
Authors: Vazquez-Ortiz Guelaguetza , Pina-Sanchez Patricia , Vazquez Karla , Duenas Alfonso , Taja Lucia , Mendoza Patricia , Garcia José A , Salcedo Mauricio
Published: July 2005

Background

Cervical carcinoma (CC) is one of the most common cancers among women worldwide and the first cause of death among the Mexican female population. CC progression shows a continuum of neoplastic transitions until invasion. Matrix metalloproteinases (MMPs) and cathepsins play a central role on the enhancement of tumor-induced angiogenesis, cell migration, proliferation, apoptosis and connective tissue degradation. MMPs -2 and -9 expression has been widely studied in cervical cancer. Nevertheless, no other metalloproteinases or cathepsins have been yet related with the progression and/or invasion of this type of cancer.

Methods

Three HPV18 CC cell lines, two HPV16 CC cell lines and three HPV16 tumor CC tissues were compared with three morphologically normal, HPV negative, cervical specimens by cDNA arrays. Overexpression of selected genes was confirmed by end point semiquantitative reverse transcription-PCR with densitometry. In situ hybridization and protein expression of selected genes was further studied by means of two tissue microarrays, one consisting of 10 HSIL and 15 CC and the other one of 15 normal cervical and 10 LSIL tissues.

Results

TIMP1, Integrins alpha 1 and 4, cadherin 2 and 11, Cathepsins F, B L2, MMP 9, 10 11 and 12 were upregulated and Cathepsin S, L, H and C, Cadherins 3 and 4, TIMP3, MMP 13, Elastase 2 and Integrin beta 8 were found to be downregulated by cDNA arrays. Endpoint RT-PCR with densitometry gave consistent results with the cDNA array findings for all three genes selected for study (CTSF, MMP11 and MMP12). In situ hybridization of all three genes confirmed overexpression in all the HSIL and CC. Two of the selected proteins were detected in LSIL, HSIL and CC by immunohistochemistry.

Conclusion

Novel undetected CC promoting genes have been identified. Increased transcription of these genes may result in overexpression of proteins, such as CTSF, MMP11 and MMP12 which could contribute to the pathogenesis of CC.
Direct microcontact printing of oligonucleotides for biochip applications
Authors: Thibault C , Le Berre V , Casimirius S , Trévisiol E , François J , Vieu C
Published: July 2005

Background

A critical step in the fabrication of biochips is the controlled placement of probes molecules on solid surfaces. This is currently performed by sequential deposition of probes on a target surface with split or solid pins. In this article, we present a cost-effective procedure namely microcontact printing using stamps, for a parallel deposition of probes applicable for manufacturing biochips.

Results

Contrary to a previous work, we showed that the stamps tailored with an elastomeric poly(dimethylsiloxane) material did not require any surface modification to be able to adsorb oligonucleotides or PCR products. The adsorbed DNA molecules are subsequently printed efficiently on a target surface with high sub-micron resolution. Secondly, we showed that successive stamping is characterized by an exponential decay of the amount of transferred DNA molecules to the surface up the 4th print, then followed by a second regime of transfer that was dependent on the contact time and which resulted in reduced quality of the features. Thus, while consecutive stamping was possible, this procedure turned out to be less reproducible and more time consuming than simply re-inking the stamps between each print. Thirdly, we showed that the hybridization signals on arrays made by microcontact printing were 5 to 10-times higher than those made by conventional spotting methods. Finally, we demonstrated the validity of this microcontact printing method in manufacturing oligonucleotides arrays for mutations recognition in a yeast gene.

Conclusion

The microcontact printing can be considered as a new potential technology platform to pattern DNA microarrays that may have significant advantages over the conventional spotting technologies as it is easy to implement, it uses low cost material to make the stamp, and the arrays made by this technology are 10-times more sensitive in term of hybridization signals than those manufactured by conventional spotting technology.
Physical health and depressive symptoms in older Europeans
Authors: Braam A. W. , Prince M. J. , Beekman A. T. F. , Delespaul P. , Dewey M. E. , Geerlings S. W. , Kivelä S.-L. , Lawlor B. A. , Magnússon H. , Meller I. , Pérès K. , Reischies F. M. , Roelands M. , Schoevers R. A. , Saz P. , Skoog I. , Turrina C. , Versporten A. , Copelan J. R. M.
Published: July 2005

Background

Associations between physical health and depression are consistent across cultures among adults up to 65 years of age. In later life, the impact of physical health on depression is much more substantial and may depend on sociocultural factors.

Aims

To examine cross-national differences in the association between physical health and depressive symptoms in elderly people across western Europe.

Method

Fourteen community-based studies on depression in later life in nine western European countries contributed to a total study sample of 22 570 respondents aged 65 years and older. Measures were harmonised for depressive symptoms (EURO-D scale), functional limitations and chronic physical conditions.

Results

In the majority of the participating samples, the association of depressive symptoms with functional disability was stronger than with chronic physical conditions. Associations were slightly more pronounced in the UK and Ireland.

Conclusions

The association between physical health and depressive symptoms in later life is consistent across western Europe.
Effectiveness of providing self-help information following acute traumatic injury: Randomised controlled trial
Authors: Turpin Graham , Downs Maria , Mason Suzanne
Published: July 2005

Background

Patients attending an accident and emergency department may exhibit psychological disturbances post-injury. Early interventions have been suggested to reduce the risk of post-injury disorder, including psychoeducation.

Aims

We assessed the efficacy of providing such self-help information.

Method

Patients who had experienced trauma were randomised to two groups: those given (n=75) and not given (n=67) a self-help booklet. Psychological assessments were completed within 2, 10–12 and 24–26 weeks.

Results

Post-traumatic stress disorder (PTSD), anxiety and depression decreased (P< 0.05) with time but there were no group differences in PTSD or anxiety. The controls were less depressed (P< 0.05) at follow-up. There was a reductionin PTSD caseness within the control (50%) compared with the intervention (20%) group which was almost significant (P< 0.06).

Conclusions

This trial failed to support the efficacy of providing self-help information as a preventive strategy to ameliorate PTSD.
Schizophrenia and suicide: Systematic review of risk factors
Authors: Hawton Keith , Sutton Lesley , Haw Camilla , Sinclair Julia , Deeks Jonathan J.
Published: July 2005

Background

Suicide risk is greatly increased in schizophrenia. Detection of those at risk is clinically important.

Aims

To identify risk factors for suicide in schizophrenia.

Method

The international literature on case-control and cohort studies of patients with schizophrenia or related conditions in which suicide was reported as an outcome was systematically reviewed. Studies were identified through searching electronic databases and reference lists, and by consulting experts.

Results

Twenty-nine eligible studies were identified. Factors with robust evidence of increased risk of suicide were previous depressive disorders (OR=3.03, 95% CI 2.06–4.46), previous suicide attempts (OR=4.09, 95% CI 2.79–6.01), drug misuse (OR=3.21, 95% CI 1.99–5.17), agitation or motor restlessness (OR=2.61, 95% CI 1.54–4.41), fear of mental disintegration (OR=12.1, 95% CI 1.89–81.3), poor adherence to treatment (OR=3.75, 95% CI 2.20–6.37) and recent loss (OR=4.03, 95% CI 1.37–11.8). Reduced risk was associated with hallucinations (OR=0.50, 95% CI 0.35–0.71).

Conclusions

Prevention of suicide in schizophrenia is likely to result from treatment of affective symptoms, improving adherence to treatment, and maintaining special vigilance in patients with risk factors, especially after losses.
Outcome of Cancer Patients Considered for Intensive Care Unit Admission: A Hospital-Wide Prospective Study
Authors: Thiéry Guillaume , Azoulay Élie , Darmon Michael , Ciroldi Magali , De Miranda Sandra , Lévy Vincent , Fieux Fabienne , Moreau Delphine , Le Gall Jean Roger , Schlemmer Benoît
Published: June 2005

Purpose

To evaluate the outcome of cancer patients considered for admission to the intensive care unit (ICU).

Patients and Methods

Prospective, one-year hospital-wide study of all cancer and hematology patients, including bone marrow transplantation patients, for whom admission to the ICU was requested.

Results

Of the 206 patients considered for ICU admission, 105 patients (51%) were admitted. Of the 101 patients who were not admitted, 54 patients (26.2%) were considered too sick to benefit, and 47 patients (22.8%) were considered to be too well to benefit from the ICU. Of these 47 patients, 13 patients were admitted later. Survival rates after 30 and 180 days were significantly associated with admission status (P < .0001). Remission of the malignancy (odds ratio [OR], 3.37; 95% CI, 1.25 to 9.07) was independently associated with ICU admission, whereas poor chronic health status (OR, 0.38; 95% CI, 0.16 to 0.74) and solid tumor (OR, 0.43; 95% CI, 0.24 to 0.78) were associated with ICU refusal. In admitted patients, 30-day and 6-month survival rates were 54.3% and 32.4%, respectively. Of the patients considered too sick to benefit from ICU admission, 26% were alive on day 30 and 16.7% on day 180. Among patients considered too well to benefit, the 30-day survival rate was a worrisome 78.7%. Calibration of the Mortality Probability Model (the only score available at triage) was of limited value for predicting 30-day survival (area under the curve, 0.62).

Conclusion

Both the excess mortality in too-well patients later admitted to the ICU and the relatively good survival in too-sick patients suggest the need for a broader admission policy.
Effects of 9-cis-Retinoic Acid on the Insulin-Like Growth Factor Axis in Former Smokers
Authors: Lee Ho-Young , Chang Yoon Soo , Han Ji-Youn , Liu Diane D. , Lee J. Jack , Lotan Reuben , Spitz Margaret R. , Hong Waun Ki
Published: June 2005

Purpose

Insulin-like growth factor (IGF) axis has been associated with the risk of lung cancer. 9-cis-retinoic acid (9-cis-RA) has shown potential chemopreventive activities in former smokers. This study was designed to evaluate the effects of 9-cis-RA on IGF axis in former smokers to identify any benefit the retinoid may have in preventing lung cancer.

Patients and Methods

Serum concentrations of IGF-I, IGF binding protein (IGFBP)-3, and their molar ratio (IGF-I/IGFBP-3) were measured with radioimmunoassay kits in stored blood samples from the participants of an original chemoprevention trial. The participants had ceased smoking for at least 12 months and were randomly assigned to receive 3 months of daily oral 9-cis-RA (100 mg) or placebo. All statistical tests were two-sided.

Results

A total of 111 samples from the study's baseline and 84 samples from the 3 months treatment were analyzed. The serum concentrations of IGF-I and IGF-I/IGFBP-3 at baseline were significantly lower in female than in male participants. After 3 months of treatment, the serum level of IGF-I and IGF-I/IGFBP-3 were significantly lower in the 9-cis-RA group than in the placebo group (P = .03 and P < .01, respectively), but the IGFBP-3 level was significantly higher (P = .03).

Conclusion

9-cis-RA treatment modulated the IGF axis in former smokers, suggesting that the IGF axis is a potential target for the chemopreventive activities of 9-cis-RA and that the serum concentrations of IGF, IGFBP-3, and IGF-I/IGFBP-3 could serve as surrogate end point biomarkers of 9-cis-RA treatment.
Prognostic Factors and Outcome of Human Herpesvirus 8–Associated Primary Effusion Lymphoma in Patients With AIDS
Authors: Boulanger Emmanuelle , Gérard Laurence , Gabarre Jean , Molina Jean-Michel , Rapp Christophe , Abino Jean-François , Cadranel Jacques , Chevret Sylvie , Oksenhendler Eric
Published: June 2005

Purpose

Primary effusion lymphoma (PEL) is a rare high-grade B-cell non-Hodgkin's lymphoma associated with Kaposi sarcoma–associated herpesvirus/human herpesvirus 8 (KSHV/HHV-8) infection, and is mostly observed in the course of HIV infection. The prognosis is poor, with reported median survival time shorter than 6 months. To date, no prognostic factor has been identified in this subset of lymphoma.

Patients and Methods

We describe here a large series of HIV-infected patients with PEL, including 28 cases diagnosed in six centers during an 11-year time period. Prognosis analysis was performed using a Cox proportional hazard regression model. Statistically significant covariates were further analyzed in a forward, stepwise multivariate model.

Results

After a median follow-up of 3.8 years (range, 10 months to 10.8 years), nine patients (32%) were still alive, and eight of them remained progression free. The median survival was 6.2 months, and the 1-year overall survival rate was 39.3%. Fourteen patients (50%) achieved complete remission, with a 1-year disease-free survival rate at 78.6%. In a multivariate analysis, only a performance status more than 2 (hazard ratio, 5.84; 95% CI, 1.76 to 19.33) and the absence of highly active antiretroviral therapy (HAART) before PEL diagnosis (hazard ratio, 3.26; 95% CI, 1.14 to 9.34) were found to be independent predictors for shorter survival.

Conclusion

Based on a retrospective series of 28 patients, two prognostic factors were identified as being independently associated with impaired clinical outcome in HIV-related PEL—(1) a poor performance status and (2) the absence of HAART before PEL diagnosis.
The Unconscious Consumer: Effects of Environment on Consumer Behavior
Authors: Dijksterhuis Ap. , Smith Pamela K. , van Baaren Rick B. , Wigboldus Daniël H.J.
Published: July 2005

In this article, we argue that consumer behavior is often strongly influenced by subtle environmental cues. Using grocery shopping as an example (or a “leitmotif,” if you wish), we first argue that the traditional perspective on consumer choice based on conscious information processing leaves much variance to be explained. Instead, we propose that many choices are made unconsciously and are strongly affected by the environment. Our argument is based on research on the perception–behavior link and on automatic goal pursuit.
Behavior Activation Is Not Enough
Authors: Janiszewski Chris , van Osselaer Stijn M.J.
Published: July 2005

We believe there is insufficient support for the direct perception‐to‐behavior and the direct goal‐to‐behavior explanations of nonconsciously mediated behavior. We propose a nonconscious behavioral choice model. We argue that behavioral choice depends on means activation, goal activation, and the relative value of a behavior for achieving a goal.
Targeted Overexpression of Leptin to Keratinocytes in Transgenic Mice Results in Lack of Skin Phenotype but Induction of Early Leptin Resistance
Authors: Rico Laura , Del Rio Marcela , Bravo Ana , Ramirez Angel , Jorcano José L. , Page M. Angustias , Larcher Fernando
Published: July 2005

The epidermis has a great potential as a bioreactor to produce proteins with systemic action. However, the consequences of ectopic epidermal protein overexpression need to be carefully addressed to avoid both local and systemic adverse effects. Thus, the long-term effects of leptin on skin physiology have not been studied, and the metabolic consequences of sustained keratinocyte-derived leptin overexpression are unknown. Herein we describe that very high serum leptin levels can be achieved from a cutaneous source in transgenic mice in which leptin cDNA overexpression was driven by the keratin K5 gene regulatory sequences. Histopathological analysis including the study of skin differentiation and proliferation markers in these transgenic mice revealed that keratinocyte-derived leptin overexpression appears not to have any impact on cutaneous homeostasis. Although young K5-leptin transgenic mice showed remarkable thinness and high glucose metabolism as shown in other leptin transgenic mouse models, a marked leptin insensitivity become apparent as early as 3–4 months of age as demonstrated by increased weight gain and insulin resistance development. Other signs of leptin/insulin resistance included increased bone mass, organomegaly, and wound healing impairment. In addition, to provide evidence for the lack of untoward effects of leptin on epidermis, this transgenic mouse helps us to establish the safe ranges of keratinocyte-derived leptin overexpression and may be useful as a model to study leptin resistance.
The Protein Kinase C Pathway Acts through Multiple Transcription Factors to Repress Gonadotropin-Releasing Hormone Gene Expression in Hypothalamic GT1–7 Neuronal Cells
Authors: Tang Qingbo , Mazur Marcus , Mellon Pamela L.
Published: July 2005

The GnRH gene uses two well-defined regions to target expression to a small population of hypothalamic GnRH neurons: a 173-bp proximal promoter and a 300-bp enhancer localized at approximately −1800 to −1500 bp from the start site. Interaction of multiple factors with the GnRH enhancer and promoter is required to confer neuron-specific expression in vivo and in cells in culture. In addition, the expression of the GnRH gene is regulated by numerous neurotransmitters and hormones. Several of these effectors act through membrane receptors to trigger the protein kinase C pathway, and 12-O-tetradecanoyl phorbol-13-acetate (TPA), a modulator of this pathway, has been shown to suppress GnRH gene expression through the promoter. We find that TPA suppresses expression through the GnRH enhancer as well as the promoter. In the enhancer, an Oct-1 binding site, a Pbx/Prep binding site, Msx/Dlx binding sites, and a previously unidentified protein-binding element at −1793, all contribute to TPA suppression. TPA treatment leads to decreased binding of Oct-1 and Pbx1a/Prep to their sites. However, a complex formed by GT1–7 nuclear extracts on the −1793 site is not affected by TPA treatment. It is known that cooperative interaction among multiple factors is necessary for GnRH gene expression; thus, one mechanism by which TPA suppresses GnRH gene expression is to disengage some of these factors from their cis-regulatory elements.
Kinetics of the MnO4− Oxidation of Anionic Surfactant (Sodiumdodecyl Sulphate): Evidence for the Formation of Soluble Colloidal MnO2
Authors: Raju  , Khan Zaheer
Published: July 2005

A conventional spectrophotometric technique was used to study the oxidation of SDS by permanganate in a perchloric acid medium. It was observed that the reaction proceeded in two stages (fast first stage followed by a relatively slow second stage). Plots of log(absorbance) versus time deviate from linearity. The kinetic and spectroscopic data are consistent with the formation of soluble colloidal MnO2. The first-order kinetics with respect to [SDS] at low concentrations shifted to second-order at higher concentrations. The kinetics of oxidation is first-order with respect to both [MnO4−] and [HClO4]. The oxidation rate was decreased by the addition of P2O74− and Mn(II) ions. Second-step oxidation is not a true path for the oxidation of the Mn(IV)–SDS reaction. In the presence of Mn(II) (a reaction product), the MnO4− oxidation of SDS becomes more complicated, and an exact dependence on [Mn(II)] can not be estimated. Different activation parameters have been evaluated. Mechanisms consistent with the kinetic data have been proposed and discussed. The –O–SO3− group is responsible for the oxidative degradation of SDS by MnO4−.
Phase Relationships of Crystalline Polymorphs of Mesogenic 4-Cyano-4′-heptyloxybiphenyl (7OCB) and 4-Cyano-4′-octyloxybiphenyl (8OCB)
Authors: Hori Kayako , Iwai Yuko , Yano Megumi , Orihara-Furukawa Reiko , Tominaga Yasunori , Nishibori Eiji , Takata Masaki , Sakata Makoto , Kato Kenichi
Published: July 2005

Temperature-dependent X-ray diffraction (XRD) studies using synchrotron radiation and Raman spectral studies in the range of 40–850 cm−1 have been carried out for crystalline polymorphs of the title compounds in order to elucidate the phase relations of the polymorphs. Transition processes between metastable phases have been confirmed with structural evidence: square-plate (SP) crystal to needle (N) crystal for 7OCB and parallelpiped plate (PP) crystal to needle (N) crystal for 8OCB; these processes were previously proposed based on the results of differential scanning calorimetry. XRD study shows that a fourth phase of 7OCB, which appears during the stabilization process and hence is referred to as an intermediate phase, is a slightly but definitely different crystalline phase from the most stable phase. SP of 7OCB stabilizes in two steps: first to the intermediate phase, which then rearranges to the most stable phase at the melting point of the intermediate phase just below the melting point of the most stable phase or on keeping at RT for several months or longer. Some structural features of the most stable phases of 7OCB and 8OCB are discussed based on the unit cell parameters derived from the powder XRD results.
H-Transfer Motions in an H-Bonded (Chloranilic Acid)-(1,3-Diazine) 1:2 Molecular Complex Studied by 35Cl NQR and 1H NMR
Authors: Ikeda Ryuichi , Takahashi Satoshi , Nihei Taka-aki , Ishihara Hideta , Ishida Hiroyuki
Published: July 2005

35Cl NQR and 1H NMR relaxation times (T1Q, T1H) were measured in a hydrogen-bonded (chloranilic acid)-(1,3-diazine) 1:2 molecular complex in a temperature range of ca. 100–300 K. NQR frequencies observed were explained by the monovalent (1−) ionic structure of chloranilic acid, implying partial H-transfer to diazine molecules. This H-transfer was shown to be dynamic by observing a single T1H minimum at ca. 120 K. Two new relaxation processes observed by NQR in high-temperature range, but undetected by NMR, were shown to be H-transfer motions between differently charged states of chloranilic acid molecules.
The Magnetic Orientation of 2-Dimensional Silver Dendrites
Authors: Katsuki Akio , Uechi Ichiro , Tanimoto Yoshifumi
Published: July 2005

A liquid/solid redox reaction between silver ion and zinc metal was investigated under a vertical and inhomogeneous magnetic field. 2-Dimensional silver dendrites produced via the reaction were drastically affected by the magnetic field. Oriented and tilted dendrites were observed under the magnetic field. A similar phenomenon was observed at a silver ion–copper metal system under a horizontal and homogeneous magnetic field. The mechanism of the phenomenon is discussed in detail.
DNA Cleavage by Good’s Buffers in the Presence of Au(III)
Authors: Tabata Masaaki , Habib Ahsan , Watanabe Keiichi
Published: July 2005

DNA (pBluescript plasmid, 2.96 kbp) cleavage by Good’s buffers, which are extensively used in laboratories of chemistry, biochemistry, or biology, was studied by gel electrophoresis, CD spectra, ESR spectra, and CV in the presence of Au(III). Incubation of plasmid DNA with Good’s buffers and Au(III) resulted in DNA cleavage. DNA cleavage occurred due to the formation of nitrogen-centered cationic free radicals from the Good’s buffers in the presence of Au(III). The formation of the nitrogen-centered free radicals was confirmed by ESR spectroscopy. Gel electrophoresis results indicated that Form I and Form II were converted into Form III and a DNA fragment. The molecular weight of the DNA fragment was estimated ca. 1.47 kbp by DNA marker gel electrophoresis. ESR spectra were not observed from the Good’s buffers in the presence of other metal ions, such as Mn(II), Fe(III), Co(II), Ni(II), Zn(II), Pd(II), Cd(II), Hg(II), and Pb(II), resulting in no DNA cleavage. Gel electrophoresis results indicated a partial DNA cleavage during the incubation of DNA in the presence of Good’s buffers and Au(III). This is because the DNA cleavage reaction stopped at 15 min, since the radical lifetime is ca. 15 min. The intensities of the CD spectra at 270 nm decreased with time by 15 min upon the addition of Au(III), and a constant CD spectrum was observed after 15 min. Further addition of Au(III) showed an alternation of the CD spectra from positive ellipticity to negative ellipticity at 270 nm, suggesting significant DNA cleavage. The mechanism of DNA cleavage is discussed based on the gel electrophoresis, ESR, CD, and CV results.
Singlet Energy Transfer and Photoinduced Electron Transfer in Star-Shaped Naphthalimide Derivatives Based on Triphenylamine
Authors: Zhu Weihong , Xu Yisheng , Zhang Yan , Shen Jianping , Tian He
Published: July 2005

A series of star-shaped naphthalimide derivatives (TPA-NP) were synthesized with a triphenylamine (TPA) core and chromophores at the rim. Their steady-state absorption, fluorescence, and transient fluorescence spectra were investigated, indicative of severely fluorescent quenching with regard to the corresponding 1,8-naphthalimide (NP) parent chromophores. Excitation into either the triphenylamine or naphthalimide absorption bands results in the occurrence of singlet energy transfer and photoinduced electron transfer (PIET) effectively. Single photon-counting experiments show a fast charge separation and a thermally activated back reaction. As a consequence, the fluorescent lifetime of the naphthalimide moiety in the system of TPA-NP is reduced by more than an order of magnitude, which mainly arises from the intramolecular PIET process of TPA and naphthalimide. The kinetics and PIET mechanism involving TPA-NP were elucidated.
An end-healing enzyme from Clostridium thermocellum with 5′ kinase, 2′,3′ phosphatase, and adenylyltransferase activities
Authors: MARTINS ALEXANDRA , SHUMAN STEWART
Published: June 2005

We identify and characterize an end-healing enzyme, CthPnkp, from Clostridium thermocellum that catalyzes the phosphorylation of 5′-OH termini of DNA or RNA polynucleotides and the dephosphorylation of 2′,3′ cyclic phosphate, 2′-phosphate, and 3′-phosphate ribonucleotides. CthPnkp also catalyzes an autoadenylylation reaction via a polynucleotide ligase-type mechanism. These characteristics are consistent with a role in end-healing during RNA or DNA repair. CthPnkp is a homodimer of an 870-amino-acid polypeptide composed of three catalytic domains: an N-terminal module that resembles the polynucleotide kinase domain of bacteriophage T4 Pnkp, a central metal-dependent phosphoesterase module, and a C-terminal module that resembles the nucleotidyl transferase domain of polynucleotide ligases. The distinctive feature of CthPnkp vis-à-vis known RNA repair enzymes is that its 3′ end modification component belongs to the calcineurin-type phosphatase superfamily. It contains putative counterparts of the amino acids that form the dinuclear metal-binding site and the phosphate-binding site of bacteriophage λ phosphatase. As with λ phosphatase, the 2′,3′ cAMP phosphatase activity of CthPnkp is specifically dependent on nickel or manganese. We identify homologs of CthPnkp in other bacterial proteomes.
Zinc-regulating Proteins, ZnT-1, and Metallothionein I/II Are Present in Different Cell Populations in the Mouse Testis
Authors: Elgazar Vered , Razanov Vladimir , Stoltenberg Meredin , Hershfinkel Michal , Huleihel Mahmoud , Nitzan Yuval Bibi , Lunenfeld Eitan , Sekler Israel , Silverman William F.
Published: July 2005

Zinc ions play an important role in testis development and spermatogenesis. Thus, nutritional zinc deficiency leads to aberrant testicular development, reduced spermatogenesis, and male sterility. The precise actions of zinc in mediating these functions and the mechanisms by which zinc is itself regulated in the testis, however, have not been adequately elucidated. We have assessed the distribution of the zinc-regulating proteins ZnT-1 and metallothionein I/II (MT I/II) in the mouse seminiferous tubule. Colabeling for ZnT-1 and MT I/II demonstrated unique patterns of distribution for these proteins, with ZnT-1 present in Sertoli cells in addition to luminal spermatozoa and MT I/II restricted to spermatocytes. These findings were confirmed by dual-label immunofluorescence for ZnT-1 and the Sertoli cell marker, vimentin, and by immunoelectron microscopy. The differential expression patterns of ZnT-1 and MTs support the hypothesis that ZnT-1 and MTs play different roles in the regulation of intracellular zinc in this organ. The specific expression of ZnT-1 in the Sertoli cells, moreover, is consistent with their role in maintaining a nurturing, closely regulated environment for spermatogenesis.
Smooth Muscle-specific α Tropomyosin Is a Marker of Fully Differentiated Smooth Muscle in Lung
Authors: Vrhovski Bernadette , McKay Karen , Schevzov Galina , Gunning Peter W. , Weinberger Ron P.
Published: July 2005

Tropomyosin (Tm) is one of the major components of smooth muscle. Currently it is impossible to easily distinguish the two major smooth muscle (sm) forms of Tm at a protein level by immunohistochemistry due to lack of specific antibodies. α-sm Tm contains a unique 2a exon not found in any other Tm. We have produced a polyclonal antibody to this exon that specifically detects α-sm Tm. We demonstrate here the utility of this antibody for the study of smooth muscle. The tissue distribution of α-sm Tm was shown to be highly specific to smooth muscle. α-sm Tm showed an identical profile and tissue colocalization with α-sm actin both by Western blotting and immunohistochemistry. Using lung as a model organ system, we examined the developmental appearance of α-sm Tm in comparison to α-sm actin in both the mouse and human. α-sm Tm is a late-onset protein, appearing much later than actin in both species. There were some differences in onset of appearance in vascular and airway smooth muscle with airway appearing earlier. α-sm Tm can therefore be used as a good marker of mature differentiated smooth muscle cells. Along with α-sm actin and sm-myosin antibodies, α-sm Tm is a valuable tool for the study of smooth muscle.
Three-dimensional Imaging of the Intracellular Localization of Growth Hormone and Prolactin and Their mRNA Using Nanocrystal (Quantum Dot) and Confocal Laser Scanning Microscopy Techniques
Authors: Matsuno Akira , Itoh Johbu , Takekoshi Susumu , Nagashima Tadashi , Osamura R. Yoshiyuki
Published: July 2005

Semiconductor nanocrystals (Quantum dots, Qdots) have recently been used in biological research, because they do not fade on exposure to light, and they enable us to obtain multicolor imaging because of a narrow emission peak that can be excited via a single wavelength of light. There have been no reports of simultaneous localization of mRNA and protein using Qdots. We successfully applied these advantages of Qdot and confocal laser scanning microscopy (CLSM) to three-dimensional images of the intracellular localization of growth hormone and prolactin and to their mRNA. In situ hybridization and immunohistochemistry using Qdots combined with CLSM can optimally illustrate the relationship between protein and mRNA simultaneously in three dimensions. Such an approach enables us to visualize functional images of proteins in relation with mRNA synthesis and localization.
Light and Electron Microscopic Immunohistochemical Detection of Bromodeoxyuridine-labeled Cells in the Brain: Different Fixation and Processing Protocols
Authors: Ngwenya Laura B. , Peters Alan , Rosene Douglas L.
Published: July 2005

Bromodeoxyuridine (BrdU) immunohistochemistry is the method of choice for labeling newly generated cells in the brain. Most BrdU studies utilize paraformaldehyde-fixed brain tissue because of its compatibility with both BrdU and other immunohistochemical methods. However, stronger fixation is required for electron microscopic studies, and unfixed tissue is needed for biochemical and molecular studies. Because there are no systematic studies comparing the effects of different fixatives on BrdU immunohistochemistry in brain tissue, we compared BrdU immunohistochemical methods in brain tissue fixed with 4% paraformaldehyde, a mixed glutaraldehyde–paraformaldehyde fixative for electron microscopy, and unfixed tissue from brains perfused only with buffer and flash frozen. After optimizing immunostaining protocols, qualitative assessments of light microscopic diaminobenzidine labeling and of double-label immunofluorescence with confocal microscopy demonstrated excellent BrdU labeling in each of the three groups. Quantitative stereological assessment of the number of BrdU-labeled cells in rat dentate gyrus showed no significant difference in the number of labeled cells detected with each perfusion protocol. Additionally, we developed a protocol to visualize BrdU-labeled cells in the electron microscope with adequate preservation of fine structure in both rat and monkey brain.
Cardiomyocyte Remodeling and Sarcomere Addition after Uniaxial Static Strain In Vitro
Authors: Yu Ji-Guo , Russell Brenda
Published: July 2005

Individual cardiomyocytes are lengthened in dilated cardiomyopathy. However, it is not known how the new sarcomeres are added to preexisting myofibrils. Using a three-dimensional microtextured culturing system, a 10% mechanical static strain was applied to aligned, well-attached cardiomyocytes from neonatal rat. The morphology of the myofibrils and the ends of the myocytes were examined. Disruptions of the sarcomeric pattern for actin showed a progression from weak to intense staining over 4 hr. The lightly stained sarcomeres were common at 1 hr after being strained, peaked at 2 hr, and then subsided. In contrast, the numbers of intensely stained sarcomeres were initially low, peaked at 3 hr, and then began to decline when compared with control values. The myocyte ends showed elongations and convolutions after 3 hr and 4 hr of mechanical strain when observed with α-actinin and N-cadherin staining. We suggest that myocytes from neonatal rat hearts remodel by insertion of new sarcomeres throughout the cell length and also by enhancement at the intercalated discs.
Immunohistochemical Determination of Cytosolic Cytochrome c Concentration in Cardiomyocytes
Authors: van Beek-Harmsen Brechje J. , van der Laarse Willem J.
Published: July 2005

Cytochrome c release from the intermembrane space of mitochondria is one of the triggers of apoptosis. There is no histochemical method available to demonstrate cytochrome c in cryostat sections, possibly because small cytosolic proteins diffuse readily into aqueous fixation media. This report shows that it is possible to demonstrate cytochrome c release in cardiomyocytes in failing myocardium using vapor fixation of cryostat sections and immunohistochemistry. The method is calibrated using sections from gelatin blocks containing known concentrations of cytochrome c. The method is applied to the hypertrophied right ventricular wall of rats in which pulmonary hypertension was induced by monocrotaline. Cytochrome c release is found in a fraction of the cardiomyocytes, leading to a mosaic-staining pattern. Cytochrome c release was found in myocytes over the full range of cross-sectional area (from 1 to 3.9 times control) in the hypertrophied myocardium. Cytosolic cytochrome c concentrations up to 0.4–0.5 mM occur frequently.