Miracle-dz/UniFAR-FTUs huggingface dataset:
Culture of Keratinocytes for Transplantation without the Need of Feeder Layer Cells

Patients with large burn wounds have a limited amount of healthy donor skin. An alternative for the autologous skin graft is transplantation with autologous keratinocytes. Conventionally, the keratinocytes are cultured with mouse feeder layer cells in medium containing fetal calf serum (FCS) to obtain sufficient numbers of cells. These xenobiotic materials can be a potential risk for the patient. The aim of the present study was to investigate if keratinocytes could be expanded in culture without the need of a feeder layer and FCS. Keratinocytes were cultured on tissue culture plastic with or without collagen type IV coating in medium containing Ultroser G (serum substitute) and keratinocyte growth factor (KGF). An in vitro skin equivalent model was used to examine the capacity of these cells to form an epidermis. Keratinocytes in different passages (P2, P4, and P6) and freshly isolated cells were studied. Keratinocytes grown on collagen type IV were able to form an epidermis at higher passage numbers than cells grown in the absence of collagen type IV (P4 and P2, respectively). In both cases the reconstructed epidermis showed an increased expression of Ki-67, SKALP, involucrin, and keratin 17 compared to normal skin. Only 50,000 keratinocytes grown on collagen type IV in P4 were needed to form 1 cm2 epidermis, whereas 150,000 of freshly isolated keratinocytes were necessary. Using this culture technique sufficient numbers of keratinocytes, isolated from 1 cm2 skin, were obtained to cover 400 cm2 of wound surface in 2 weeks. The results show that keratinocytes can be cultured without the need of a fibroblast feeder layer and FCS and that these cells are still able to create a fully differentiated epidermis. This culture technique can be a valuable tool for the treatment of burn wounds and further development of tissue engineered skin.
Human Milk Oligosaccharides Protect Against Enteropathogenic Escherichia coli Attachment In Vitro and EPEC Colonization in Suckling Mice

ABSTRACT Breast-feeding reduces the risk of enteric bacterial infections in newborns in part because of human milk oligosaccharides (HMOs), complex glycans that are present in human milk, but not in infant formula. Enteropathogenic Escherichia coli (EPEC) are attaching/effacing pathogens that cause serious diarrheal illness with potentially high mortality in infants. We isolated HMOs from pooled human milk and found that they significantly reduce EPEC attachment to cultured epithelial cells. In suckling mice, administration of HMOs significantly reduced colonization with EPEC compared with untreated controls. These data suggest an essential role for HMOs in the prevention of EPEC infections in human infants.
Java project on periodontal diseases: serotype distribution of Aggregatibacter actinomycetemcomitans and serotype dynamics over an 8-year period.

OBJECTIVE To investigate the serotype distribution and stability of Aggregatibacter actinomycetemcomitans over an 8-year period in untreated Indonesian subjects. MATERIAL AND METHODS Clinical periodontal status and the presence of A. actinomycetemcomitans were established in 1994 and 2002 in 107 subjects from an Indonesian tea estate deprived from dental care. On an average, 3.6 isolates per patient were subcultured and serotyped using specific PCR reactions. RESULTS In 1994, the predominant serotype was b (53.7%), whereas a and c occurred in 17.1% and 14.6% of the subjects, respectively. In 2002, a reduction in serotypes a (7.5%) and b (30.2%) occurred. Serotypes c and e increased in prevalence from 14.6% to 35.8% and 2.4% to 9.4%, respectively. Multiple serotypes were found in 12.2% in 1994 and 17% in 2002. From 24 subjects who were positive at both time points, 14 (58.3%) had the same serotype, whereas in 10 subjects (41.7%), a different serotype was found. Mean clinical attachment loss had increased from 0.74 mm in 1994 to 1.96 mm in 2002 but could not be related to subgingival presence of A. actinomycetemcomitans. CONCLUSIONS A. actinomycetemcomitans serotypes distribution in Indonesian young adults shifts from predominantly serotype b to a more equal prevalence of serotypes b and c. This shift suggests an opportunistic character of A. actinomycetemcomitans.
The mismatch between high effort and low reward in household and family work predicts impaired health among mothers.

BACKGROUND So far, Siegrist's model of effort-reward imbalance (ERI) has been tested almost exclusively for paid employment. This article reports results on a newly developed questionnaire measuring ERI in unpaid household and family work. METHODS Using data of a population-based sample of 3129 German mothers, logistic regression analyses were performed to test the following three main assumptions: (i) high effort combined with low reward in household and family work increases the risk of poor health; (ii) a high level of overcommitment may enhance the risk of poor health; and (iii) mothers reporting an extrinsic high ERI and a high level of overcommitment have an even higher risk of poor health. RESULTS ERI was significantly related to self-rated health, somatic complaints and mental health. A high level of overcommitment increased the risk of poor health, whereas ERI and overcommitment combined was associated with the highest risk of poor health. Statistically significant synergy effects of combined exposure of ERI and overcommitment were found for 'anxiety'. CONCLUSIONS With some limitations, all three assumptions underlying the ERI model were confirmed. Thus, we conclude that ERI is applicable to domestic work and may provide an explanatory framework to assess stress experiences in mothers.
Safety of an intravitreal injection of triamcinolone: results from a randomized clinical trial.

OBJECTIVE To determine the safety of a single intravitreal injection of triamcinolone acetonide (4 mg) in patients with subfoveal choroidal neovascularization caused by age-related macular degeneration. METHODS A double-masked, placebo-controlled, randomized clinical trial was conducted at a public tertiary referral eye hospital. Patients participating had age-related macular degeneration with evidence of choroidal neovascularization, any part of which was classic; age older than 59 years; and best-corrected visual acuity of 20/200 or better. Eyes were assigned to active study treatment or to placebo. Intraocular pressure and cataract grading were performed every 6 months for 3 years. Adverse events, from mild to vision-threatening or life-threatening, were recorded as procedure-related or corticosteroid-related. RESULTS Seventy-five eyes were assigned to study treatment and 76 eyes to placebo. There were no moderate or severe adverse events related to the surgical procedure in either group. Triamcinolone-treated eyes had a significantly increased risk of developing mild or moderate elevation of the intraocular pressure. Topical glaucoma medication reduced intraocular pressure to acceptable levels in all patients. There was significant progression of cataract in the triamcinolone-treated eyes. CONCLUSION Despite a significant adverse event profile, intravitreal triamcinolone is generally well tolerated by the human eye as long as patients are carefully followed up by their surgeon and treated appropriately, when necessary.
Early signal transduction by the antigen receptor without commitment to T cell activation.

The T lymphocyte antigen-receptor complex mediates antigen-specific cell activation, at least in part, through the production of inositolphospholipid-derived second messengers. Little is known about how second messenger events, typically measured within minutes of ligand binding, eventually lead to distal biologic responses such as expression of lymphokine genes. Several monoclonal antibodies directed against the receptor complex were tested for their ability to elicit transmembrane signaling in the parental Jurkat line and in a somatic mutant (J.CaM1) with a deficient receptor function. One antibody elicited substantial early Ca2+ mobilization responses in both cells but was unable to promote expression of the interleukin-2 gene in J.CaM1. In J.CaM1 there was a diminished production of phosphatidylinositol second messengers, and the elevation in intracellular free Ca2+ was transient. Thus, short-term Ca2+ mobilization does not always indicate complete signal transmission and lead to a full cellular response.
The p53 Family Member Genes Are Involved in the Notch Signal Pathway*

The p53 tumor suppressor is a transcription factor that regulates cell growth and death in response to environmental stimuli such as DNA damage. p63/p51 andp73 were recently identified as members of the p53gene family. In contrast to p53 however, p63and p73 are rarely mutated in human cancers. Mice that lack p53 are developmentally normal, while p63 and p73 appear to play critical roles in normal development. To determine how p63 and p73 are involved in normal development, we attempted to identify target genes that are specifically regulated by p63 and/or p73 but not by p53. We found that the Jagged1 (JAG1) andJagged2 (JAG2) genes, encoding ligands for the Notch receptors, are up-regulated by p63 and p73. Furthermore, we identified a p63-binding site in the second intron of the JAG1gene, which can directly interact with the p63 protein in vivo, as assessed by a chromatin immunoprecipitation assay. A heterologous reporter assay revealed that this p63-binding site is a functional response element and is specific for p63. We also found a target of Notch signaling, HES-1 was up-regulated in Jurkat cells, in which Notch1 is highly expressed, when co-cultured with p63-transfected cells, suggesting that p63 can trigger the Notch signal pathway in neighboring cells. Our findings show an association between the p53 family genes and Notch signaling and suggest a potential molecular mechanism for the involvement of the p53family genes in normal development.
Activation of human immunodeficiency virus type 1 in monocytoid cells by the protozoan parasite Leishmania donovani

In this study, we demonstrated that the protozoan parasite Leishmania donovani and one of its major surface molecules, the lipophosphoglycan (LPG), can induce human immunodeficiency virus type 1 (HIV-1) expression in U1 and OM-10.1, two cell lines of monocytoid origin latently infected with HIV-1. Treatment of U1 cells with various concentrations of LPG (1, 5, and 10 microM) resulted in a dose-dependent secretion of tumor necrosis factor alpha (TNF-alpha). Suppression of LPG-induced HIV-1 expression by polyclonal anti-TNF-alpha antibodies further confirmed the involvement of this cytokine. Results from these studies indicate that the protozoan parasite L. donovani can induce the secretion of TNF-alpha that will function in an autocrine or paracrine manner to upregulate HIV-1 expression. Our data suggest for the first time that this protozoan parasite can be viewed as a potential cofactor in the pathogenesis of AIDS.
Comparison of Two Dosages of Ginkgo biloba Extract EGb 761 in Patients with Peripheral Arterial Occlusive Disease Fontaine’s Stage IIb

Summary Background: Rheologically-active drugs are widely used in the therapy of peripheral arterial occlusive disease (pAOD) Fontaine’s stage II. Several clinical trials have demonstrated the efficacy of Ginkgo biloba extract in the treatment of pAOD Fontaine’s stage II. A pilot study indicated the superiority of 240 mg Ginkgo biloba extract daily compared with the standard dosage of 120 mg to 160 mg daily. This trial was conducted to confirm the superiority of the higher dosage of Ginkgo biloba extract in patients with pAOD Fontaine’s stage IIb statistically. Patients and methods: 74 patients were analysed. Thirty-eight patients received the standard dosage (EGb 761 120 mg/d), and 36 patients received 240 mg EGb 761 daily. The primary efficacy criterion was the difference of the pain-free walking distance between the start of treatment and after 24 weeks measured on a treadmill under standardised conditions. Results: The pain-free walking distance improved in both groups. There was a mean increase of 60.6 m in the group of patients who received 120 mg Ginkgo biloba extract daily and a statistically significant higher (p = 0.0253) mean increase of 107.0 m in the group of patients who were treated with the higher dosage. Conclusion: Both dosage regimens investigated in this trial led to a clinically relevant improvement of the pain-free walking distance after 24 weeks of treatment. The superiority of the higher dosage over the standard dosage was statistically significant. Both treatment variations were safe and well tolerated.
Initial features and outcome of cutaneous and non‐cutaneous primary extranodal follicular lymphoma

Follicular lymphoma (FL), a typically nodal disease, can arise in extranodal sites in about 10% of cases. The present study aimed to analyse the main differential features of patients with primary extranodal FL. Thirty‐nine patients with primary extranodal FL were identified from a series of 354 patients with FL diagnosed at a single institution and their main clinicobiological features were analysed. Twenty patients (5·6%) had a primary extranodal non‐cutaneous FL, and 19 (5·4%) a cutaneous FL. BCL2+ and CD10+ expression and BCL2/IGHJ@ rearrangement were less frequently observed in cutaneous FL. Absence of ‘B’‐symptoms, early stage, absence of bone marrow involvement and low‐risk Follicular Lymphoma International Prognostic Index (FLIPI) were more frequent in extranodal FL. Five‐year overall survival (OS) was 100%, 83% and 78% for cutaneous, non‐cutaneous and nodal FL, respectively. When stage I patients were analysed separately, no differences were seen in terms of OS. In multivariate analysis, FLIPI was the most important variable to predict outcome. In conclusion, extranodal FLs, particularly cutaneous, have particular clinico‐biological features, which differentiate them from nodal cases. Nevertheless, primary site of the disease is not the main issue to predict outcome.
Multiple representations of body movements in mesial area 6 and the adjacent cingulate cortex: An intracortical microstimulation study in the macaque monkey

The mesial agranular frontal cortex that lies rostral to area 4 (F1) is formed by two distinct cytoarchitectonic areas: F3, located caudally, and F6, located rostrally. In the present experiments we investigated the organization of F3 and F6 by observing the motor responses evoked by their intracortical electrical microstimulation. Our main purpose was to find out whether the cytoarchitectonic subdivision of the mesial agranular frontal cortex into two areas has a physiological counterpart.
Effectiveness of bougie dilation for the management of corrosive esophageal strictures.

BACKGROUND AND STUDY AIMS Caustic ingestion caused by swallowing a detergent can produce a progressive and devastating injury in the esophagus and stomach. One of the most important outcomes of the corrosive oesophagitis is the stricture formation, which is resistant to treatment. The aim of this study was firstly to determine the relation between agent, inflammation and stricture, and secondly investigate the efficiency of dilation in patients having esophageal stricture due to corrosive oesophagitis. PATIENTS AND METHODS In this study, 58 cases with post caustic oesophagitis, which had been admitted to our clinic or emergency department between January 1999 and December 2004, were assessed retrospectively. Dilation of esophageal stricture of the cases was performed by Savary-Gilliard bougies. RESULTS The most frequently ingested substance was alkaline (48.2%). Concerning all the patients, the most frequent location of caustic injury was upper esophagus (36.2%), and grade I injury was the most frequently encountered one (34.4%). Thirty patients (51.7%) developing stricture were treated by repeated dilations. The most common location of stricture was middle esophagus (50%), and severe stricture was the most common one among all stricture grades (46.7%). Alkaline ingestions yielded more severe stricture than acids. Eight of the patients with stricture (26.6%, 8/30), who didn't respond to periodic esophageal dilation, underwent esophageal resection or bypass surgery. CONCLUSION Dilation with Savary-Gilliard bougies is a quite effective method for stricture after corrosive oesophagitis.
Naturally occurring sites within the Shigella dysenteriae tryptophan operon severely limit tryptophan biosynthesis

We investigated the structural, functional, and regulatory properties of the Shigella dysenteriae tryptophan (trp.) operon in transduction hybrids in which the cysB-trp-region of Escherichia coli is replaced by the corresponding region from S. dysenteriae. Tryptophan biosynthesis was largely blocked in the hybrids, although the order of the structural genes was identical with that of E. coli. Nutritional tests and enzyme assays revealed that the hybrids produced a defective anthranilate synthetase (ASase). Deletion mapping identified two distinct sites in trpE, each of which was partially responsible for the instability and low activity of ASase. We also discovered a pleiotropic site trpP (S) that maps outside the structural gene region and is closely linked to the S. dysenteriae trp operator. trpP (S) reduced the rate of trp messenger ribonucleic acid synthesis, and consequently trp enzyme levels, 10-fold relative to wild-type E. coli. In recombinants in which the structural genes of E coli were under the control of the S. dysenteriae promoter, enzyme levels were also reduced 10-fold. In some fast-growing revertants of the original hybrids, the rates of trp messenger ribonucleic acid synthesis and levels of tryptophan synthetase were restored to values characteristic of wild-type E.coli. Thus, the Trp auxotrophy associated with the S dysenteriae trp operon derives from the combination of a defective ASase and decreased expression of the entire operon imposed by trpP (S).
Effects of androgen deprivation on glycaemic control and on cardiovascular biochemical risk factors in men with advanced prostate cancer with diabetes

Introduction and objective. Androgen deprivation therapy of prostate cancer with luteinizing hormone releasing hormone agonists may result in loss of bone mass, changes in body composition and a deterioration of arterial stiffness. The present study monitored the effects of androgen deprivation therapy in men with insulin-dependent diabetes on glycaemic control and on biochemical cardiovascular risk markers. Methods. Twenty-nine patients from a urology practice were included. All men had insulin-dependent diabetes mellitus prior to being diagnosed with metastatic prostate cancer. In a retrospective analysis, levels of fasting glucose, haemoglobin A1c, insulin requirements, total cholesterol, HDL, LDL, triglycerides, fibrinogen, PAI-1, tPA and C-reactive protein were obtained on at least eight occasions over a period of up to 24 months. Results. Glycaemic control worsened substantially with increases of serum glucose requiring increases in insulin dosages. HbA1c levels rose indicating impaired glycaemic control. All biochemical cardiovascular risk markers deteriorated. Conclusion. In men with insulin-dependent diabetes, androgen deprivation therapy may have negative effects on their glycaemic control and may aggravate the biochemical risk profile of cardiovascular disease to which diabetics are predisposed. These observations are in agreement with the emerging role of low levels of testosterone in metabolic syndrome and insulin resistance.
American Ginseng Improves Glycemia in Individuals with Normal Glucose Tolerance: Effect of Dose and Time Escalation

Objective: We studied the effect of escalating the dose and administration time of American ginseng (AG, Panax quinquefolius L.) in nondiabetic individuals to achieve further improvements in glucose tolerance seen previously when 3g of AG was taken 40 minutes before a 25g glucose challenge. Methods: Ten nondiabetic individuals (6M:4F; mean ± STD: age = 41 ± 13 years, BMI = 24.8 ± 3.5 kg/m2, FBG = 4.5 ± 0.1mmolL−1) on 12 separate occasions, randomly received 0 (placebo), 3, 6 or 9g of ground AG root at 40, 80, or 120 minutes before a 25g oral glucose challenge. Capillary blood glucose was measured prior to ingestion of AG or placebo capsules and at 0, 15, 30, 45, 60 and 90 minutes from start of challenge. Results: Compared with the placebo, 3, 6 and 9g of AG reduced (p<0.05) postprandial incremental glucose at 30, 45 and 60 minutes; also, 3 and 9g of AG did so at 90 minutes. At 60 minutes, 9g of AG reduced incremental postprandial glucose relative to 3g of AG (p<0.05). All AG doses reduced (p<0.05) area under the incremental glucose curve (3g, 26.6%; 6g, 29.3%; 9g, 38.5%). AG taken at different times did not have an additional influence on postprandial glycemia. Conclusions: In nondiabetic individuals, 3, 6 or 9g of AG taken 40, 80 or 120 minutes before a glucose challenge similarly improved glucose tolerance.
Association of serology with the endovascular presence of Chlamydia pneumoniae and cytomegalovirus in coronary artery and vein graft disease.

BACKGROUND Chemotherapeutic treatment for patients with symptomatic coronary artery disease to reduce cardiovascular events may be initiated in response to elevated antibody titers against Chlamydia pneumoniae or cytomegalovirus. How antibody titers are associated with the endovascular presence of these microorganisms is still unclear. METHODS AND RESULTS Antibody titers against C pneumoniae (microimmunofluorescence) and cytomegalovirus (ELISA) in patients undergoing primary (coronary desobliterates, n=80) or repeated CABG (occluded vein grafts, n=45) were correlated with the endovascular presence of the 2 microorganisms. C pneumoniae was detected by means of a nested polymerase chain reaction (PCR) and by culturing. Both conventional PCR and quantitative PCR were applied for detection of cytomegalovirus. C pneumoniae (PCR/culture) was detected in 19/9% (15/80 and 7/80) of coronary desobliterates and in 18/11% (8/45 and 5/45) of occluded vein grafts. There was no statistical evidence that IgG values differed between patients with or without C pneumoniae detection who were undergoing primary CABG. In contrast, repeated-CABG patients with a positive PCR (P=0.0027) or C pneumoniae culture (P=0.0018) had distinctly elevated IgG titers compared with patients in whom C pneumoniae was not detected. Cytomegalovirus could not be detected in the examined specimens. CONCLUSIONS Cytomegalovirus infection does not seem to be associated with advanced coronary artery lesions. C pneumoniae antibody titers are not associated with the endovascular presence of C pneumoniae in patients with coronary artery disease. The observed strong association between elevated IgG titers and the detection of C pneumoniae in occluded vein grafts warrants further investigation.
The obesity–asthma link in different ages and the role of Body Mass Index in its investigation: findings from the Genesis and Healthy Growth Studies

To date, an obesity/asthma link is well defined in adults; however, the nature of such a link is obscure in children, partly due to Body Mass Index (BMI) limitations as a surrogate fat mass marker in childhood. We thus opted to investigate the association of adiposity with asthma in children of different ages, using several indices to assess fat mass.