bluuebunny/crossref_metadata_2025_split huggingface dataset:
Possibility of Undifferentiated Human Thigh Adipose Stem Cells Differentiating into Functional Hepatocytes
Authors: Lee Jong Hoon , Lee Kuk Han , Kim Min Ho , Kim Jun Pyo , Lee Seung Jae , Yoon Jinah 
Published: November 2012

Background This study aimed to investigate the possibility of isolating mesenchymal stem cells (MSCs) from human thigh adipose tissue and the ability of human thigh adipose stem cells (HTASCs) to differentiate into hepatocytes.
Methods The adipose-derived stem cells (ADSCs) were isolated from thigh adipose tissue. Growth factors, cytokines, and hormones were added to the collagen coated dishes to induce the undifferentiated HTASCs to differentiate into hepatocyte-like cells. To confirm the experimental results, the expression of hepatocyte-specific markers on undifferentiated and differentiated HTASCs was analyzed using reverse transcription polymerase chain reaction and immunocytochemical staining. Differentiation efficiency was evaluated using functional tests such as periodic acid schiff (PAS) staining and detection of the albumin secretion level using enzyme-linked immunosorbent assay (ELISA).
Results The majority of the undifferentiated HTASCs were changed into a more polygonal shape showing tight interactions between the cells. The differentiated HTASCs up-regulated mRNA of hepatocyte markers. Immunocytochemical analysis showed that they were intensely stained with anti-albumin antibody compared with undifferentiated HTASCs. PAS staining showed that HTASCs submitted to the hepatocyte differentiation protocol were able to more specifically store glycogen than undifferentiated HTASCs, displaying a purple color in the cytoplasm of the differentiated HTASCs. ELISA analyses showed that differentiated HTASCs could secrete albumin, which is one of the hepatocyte markers.
Conclusions MSCs were islolated from human thigh adipose tissue differentiate to heapatocytes. The source of ADSCs is not only abundant abdominal adipose tissue, but also thigh adipose tissue for cell therapy in liver regeneration and tissue regeneration.
Usefulness of V-Y Advancement Flap for Defects after Skin Tumor Excision
Authors: Kwon Ki Hyun , Lee Dong Gwan , Koo Su Han , Jo Myoung Soo , Shin Heakyeong , Seul Jung Hyun 
Published: November 2012

Background After skin tumor excision on the face, extremities, or trunk, the choice of treatment for a skin defect is highly variable. Many surgeons prefer to use a local flap rather than a skin graft or free flap for small- or moderately-sized circular defects. We have used unilateral or bilateral V-Y advancement flaps, especially on the face. Here we evaluated the functional and aesthetic results of this technique.
Methods All of the patients were pathologically diagnosed with squamous cell carcinoma (SCC), basal cell carcinoma (BCC), or malignant melanoma or premalignant lesion (Bowen's disease). Thirty-two patients underwent V-Y advancement flap repair (11 unilateral and 21 bilateral) from January 2007 to June 2011. We analyzed the patients' age and satisfaction, and location and size of defect. The patients were followed up for 6 months or more.
Results There were 22 women and 10 men. The ages ranged from 47 to 93 years with a mean age of 66 years. The causes were SCC in 15 cases, BCC in 13 cases, malignant melanoma in 1 case, Bowen's disease in 2 cases, and another cause in 1 case. The tumor locations were the face in 28 patients, and the scalp, upper limb, and flank each in one patient. All of the flaps survived and the aesthetic results were good. Postoperative recovery was usually rapid, and no complication or tumor recurrence was observed.
Conclusions The V-Y advancement flap is often used not only for facial circular defects but also for defects of the trunk and extremities. Its advantages are less scarring and superior aesthetic results as compared with other local flap methods, because of less scarification of adjacent tissue and because it is an easy surgical technique.
Efficacy of long‐term osteopenia/osteoporosis treatment in the Slovene HIV‐infected male population
Authors: Maver M , Vovko T , Pfeifer M , Volcansek G , Pecavar B , Tomazic J 
Published: November 2012

Purpose of studyIn our study from 2006, high prevalence of reduced bone mineral density (BMD) among Slovene HIV‐infected male population was observed. The aim of the present study was to evaluate bone status six years after treatment intervention.Methods69 HIV‐infected male patients (out of 96, studied in 2006) were assessed for change in BMD using dual X‐ray absorptiometry and markers of bone turnover. The effects of HIV‐associated factors (time since infection, ART duration, viral load, CD4+count), changes in life‐style risk factors (smoking, physical activity, alcohol consumption, amount of milk in diet) and treatment of osteopenia/osteoporosis were assessed. Therapy for osteopenia/osteoporosis was used in 30 patients; vitamin D and calcium supplementation in 20 patients with osteopenia and additionally bisphophonates in 15 patients with osteoporosis or androgens in case of hypogonadism, respectively. 39 patients were not treated.Summary of resultsTherapy of osteopenia/osteoporosis maintained BMD values in 18 (66.7%) patients, as compared to only 8 (25%) patients without therapy (p=0.002). Lumbar spine BMD increased by 4.1% during the 6‐year period for patients with therapy, as compared with an increase of 0.9% for patients without therapy (p=0.033). The difference in total hip values was even more significant, with a 2.6% increase in BMD with therapy, compared to a 1.95% decrease without therapy (p<0.0001). Some increase in the physical activity and milk consumption among HIV‐infected population was observed, while the smoking rate and alcohol consumption remained the same. Risk‐factors associated with life‐style did not have any effect on BMD change. According to the logistic regression model, long‐term vitamin D and calcium supplementation was the most significant factor associated with maintaining BMD values (p=0.010; OR= 14.4).ConclusionsLong‐term treatment, especially with vitamin D and calcium, was safe and efficient in the treatment of osteopenia/osteoporosis in HIV infected patients. As BMD values decreased in a majority of individuals without therapy, and given the absence of any association between life‐style change and BMD improvement, early vitamin D and calcium supplementation in the HIV‐infected population is required.
Therapeutic drug monitoring (TDM) of atazanavir in pregnancy
Authors: Else L , Jackson V , Brennan M , Breiden J , Lawless M , Coulter Smith S , Back D , Khoo S , Lambert J 
Published: November 2012

Purpose of the studyPregnant women experience physiological changes during pregnancy resulting in clinically significant alterations in antiretroviral pharmacokinetics (PK). Therefore, achieving and maintaining optimal plasma concentrations of antiretroviral drugs is essential for maternal health and minimising the risk of mother‐to‐child transmission of HIV. The aim of this study is to describe atazanavir/ritonavir (ATV/r) PK during pregnancy.MethodsPregnant HIV‐positive women received ATV/r as part of their routine pre‐natal care. Demographic and clinical data were collected, and ATV plasma concentrations [ATV] were determined in the first (T1), second (T2) and third (T3) trimester using HPLC‐MS/MS (LLQ=0.05 µg/mL). Postpartum (PP) sampling was performed where applicable. Antepartum (AP) and PP PK parameters were compared using a one‐way ANOVA.Summary of resultsFrom January 2007, 44 women (37 black African) were enrolled in the study. All received ATV/r at a standard dose of 1 tablet once daily (300/100 mg od). 24 women were receiving ART prior to pregnancy, and 20 women initiated ATV/r during pregnancy. Median (range) gestation at treatment initiation in these patients was 23.5 weeks (7–35). At the time nearest to delivery 31 patients had an undetectable plasma viral load (pVL), 6 patients had detectable pVL and 2 were unavailable. [ATV] were determined in 11/44 (T1); 25/44 (T2); 35/44 (T3) and 28/44 (PP) patients. Time of TDM sampling, gestation time and [ATV] (geometric mean; 95% CI) are given in the Table. 6 patients were either below or approaching the ATV MEC (0.15 µg/mL) during pregnancy; of these, 4/6 achieved undetectable pVL at the time of delivery (1=pVL of 291 copies/mL; 1 unavailable). [ATV] were significantly lower at T2/T3 relative to T1/PP. Equally, in a paired analysis of 28 patients (T2/T3 vs. PP), [ATV] were significantly reduced at T2/T3 (P=0.003).

T1 (n=11)
T2 (n=25)
T3 (n=35)
PP (n=28)
P value

[ATV], µg/mL
1.07 (0.15–1.99)
0.68 (0.39–0.97)
0.63 (0.47–0.78)
1.22 (0.96–1.49)
0.002

CV, %
102
83
61
49
–

Time of sampling, h
12.0 (8.8–15.3)
18.2 (15.8–20.5)
18.9 (17.2–20.7)
18.9 (16.5–21.3)
0.003

Gestation/PP, weeks
9 (6–12)
20 (14–26)
32 (25–39)
10.5 (5–50)
–

ConclusionsThis study represents one of the larger cohorts of women undergoing TDM for ATV in pregnancy. Lower [ATV] were seen in T2 and T3 when compared to T1. However, such findings were not associated with viral breakthrough or HIV transmissions. Nonetheless, careful monitoring of women in pregnancy is required, and if there is concern for inadequate levels, dose adjustment of ATV upward from 300 mg to 400 mg may be an option.
Development and validation of a LC‐MS/MS assay to quantify intracellular tenofovir‐diphosphate (TFV‐DP) and emtricitabine‐triphosphate (FTC‐TP)
Authors: Watson V , Else L , Tjia J , Khoo S , Back D 
Published: November 2012

IntroductionCombination antiretroviral therapy has been associated with dramatic reductions in morbidity and mortality in HIV‐infected patients. The key to successful HIV treatment is strict adherence to the prescribed regimen. The combination of tenofovir, emtricitabine and efavirenz co‐formulated in a single tablet (Atripla) provides a ‘one tablet once a day’ regimen, making adherence easier for patients. Tenofovir and emtricitabine are converted intracellularly to active phosphate anabolites. Therefore, knowledge of intracellular pharmacokinetics should aid understanding of the virological response and toxicity of these agents. This intracellular methodology has been developed and applied to clinical trial samples.MethodPeripheral blood mononuclear cells (PBMC) were isolated from whole blood by gradient centrifugation using ficoll. PBMC were counted and lysed (70% MeOH) giving a final cell density of 2×106 cells/ml. Lysate was centrifuged, supernatant (intracellular matrix), was spiked with drug anabolites at various concentrations to produce standard curves and quality controls (QC). Samples were extracted by protein precipitation (acetonitrile), evaporated to dryness, internal standard (IS) added, and reconstituted in 5 mM ammonuim formate. Weak anion exchange chromatography with an optimised step‐wise gradient, interfaced with a triple quadrupole mass spectrometer operated in SRM with positive ionisation mode was used for quantification.ResultsAnalytes eluted within 12 minutes run‐time with adequate separation. Calibration curves were validated over the following range TFV‐DP = 0.35–10.91 ng/mL, FTC‐TP = 0.38–103.17 ng/mL (r2 values > 0.99; linear 1/x). The lower limit of quantification was < 20%, signal to noise was > 5% and carryover < 0.1%. The precision (relative standard deviation,% RSD) of the assays as determined from analysis of quality control samples was; TFV‐DP = 6.3–11% and FTC‐TP = 6–18.6%, and accuracy was TFV‐DP = 97.5–100.8%, FTC‐TP = 98–100.3%.ConclusionsA direct, highly sensitive intracellular anabolite assay has been developed and validated which elutes analytes and IS rapidly and has been applied to patient samples from a pivotal trial the results of which are crucial in our understanding of antiretroviral drug ‘forgiveness’.
GAM analysis of the relationship between DRV PK and pharmacodynamics following DRV/r 800/100 mg qd in the phase III trials ARTEMIS and ODIN
Authors: Kakuda T , Tomaka F , Van De Casteele T , Vangeneugden T 
Published: November 2012

BackgroundOnce‐daily (qd) darunavir/ritonavir (DRV/r) 800/100mg was evaluated in two randomised and controlled Phase III trials, showing high response rates in treatment‐naïve (ARTEMIS; TMC114‐C211) and treatment‐experienced patients with no DRV resistance‐associated mutations (ODIN; TMC114‐C229). Treatment interruptions, medical co‐morbidities, drug‐drug interactions, and challenges to adherence can decrease exposures and result in suboptimal DRV coverage. Consequences of suboptimal dosing could be detrimental to patients, namely the selection of drug‐resistant HIV strains. No relevant relationship between DRV area under the plasma concentration‐time curve over 24 hours (AUC24h) or plasma trough concentrations (C0h) and efficacy or safety were observed in ARTEMIS or ODIN [1,2]. The PK/PD analysis for efficacy has been extended using a generalized additive model (GAM).MethodsA univariate GAM analysis was undertaken for each trial, exploring the relationship between DRV C0h and virological response at Week 48. A quadratic spline (2 degrees of freedom) was selected, based on the Akaike information criterion value. Additionally, a linear logistic regression model was applied to model virologic response as a function of DRV C0h.ResultsIn ARTEMIS, there was no relationship between DRV C0h and the observed virological response or virological response predicated in the GAM analysis, with no indication that lower values within the observed DRV C0h range are associated with lower predicted virological response (Figure A). The mean predicted virological response was 92.8% in the total population (N = 304). A 50% reduction in DRV C0h would result in a similar predicted mean virological response (93.9%). In ODIN, for values of DRV C0h up to 3000ng/mL, there was no clear relationship between DRV C0h and observed virological response. The GAM analysis showed a counter‐intuitive inverse relationship between DRV C0h and predicted virological response (Figure B). The mean predicted virological response was 80.6% in the total population (N = 252). For each trial, similar results to GAM were obtained when a linear logistic regression model was applied.ConclusionIn patients who received 800/100mg qd DRV/r in the ARTEMIS and ODIN trials, a reduction in DRV C0h of up to 50% is not associated with a reduced virologic response at Week 48, as predicted by both the GAM and logistic regression model. DRV/r doses below 800/100mg qd should not be administered.image
Etravirine concentrations in seminal plasma in HIV‐infected patients
Authors: Tiraboschi J , Niubo J , Ferrer E , Barrera‐Castillo G , Rozas N , Maso‐Serra M , Podzamczer D 
Published: November 2012

Purpose of the studyGood penetration of antiretroviral drugs to the seminal plasma may be associated with a decrease in viral replication and play an important role in the prevention of sexual transmission of HIV. We present data from a series of HIV‐infected ARV‐experienced patients receiving etravirine‐containing regimens, in whom etravirine concentrations and viral loads were determined in blood plasma and seminal plasma. The objective was to determine etravirine concentrations and HIV‐1 viral load (VL) in blood plasma (BP) and seminal plasma (SP) of HIV‐infected patients.MethodsTen HIV‐1 adult antiretroviral‐experienced patients receiving an etravirine‐containing regimen for at least 1 month were enrolled. Semen and blood samples were both collected around 12–24 h after the last etravirine dose, depending on once‐daily or twice‐daily dosing, respectively. HPLC/MS/MS was used to determine etravirine concentrations, and HIV‐1 VL was determined by real‐time PCR (limit of detection, VL 40 copies/mL).ResultsTen blood and twenty semen samples were collected. Median (range) CD4 count was 502 cells/mm3 (252–817) and median (range) BP VL was<40 copies/mL (40–362). Median (range) time on etravirine was 52 weeks (12–124). Median (range) BP etravirine concentration was 452.5 ng/mL (258–751). Median (range) SP etravirine concentration was 62.9 ng/mL (31.2–166), and values were above the protein‐free IC50 range (0.39–2.4 ng/mL) in all cases. Median (range) etravirine SP:BP ratio was 0.16 (0.07–0.26). SP VL was<40 copies/mL in all patients, whereas BP VL was detectable in one patient with poor adherence to treatment.

Patient
SP (ng/ml)
BP (ng/ml)
SP VL (copies/ml)
BP VL (copies/ml)
SP:BP
Concomitant ARV

1
31.2
375
<40
<40
0.08
DRVr/TDF

2*
57.1
307
<40
<40
0.18
3TC/ABC

3*
36.9
506
<40
<40
0.07
TDF/FTC

4*
60.6
399
<40
<40
0.15
3TC/ABC

5
68.1
258
<40
<40
0.26
LPVr/TDF

6*
104
414
<40
<40
0.25
3TC/ABC

7
107
491
<40
362
0.21
DRVr/TDF/FTC

8*
65.3
592
<40
<40
0.11
TDF/FTC

9
52.2
518
<40
<40
0.1
DRVr

10
166
751
<40
<40
0.22
TDF/FTC

Median
62.9
452.5
<40
<40
0.16

Patients taking ETR once a day.

ConclusionsTotal etravirine concentrations in male genital secretion are modest, reaching only 16% of the BP concentration, but nevertheless, more than 10 times above the wild type IC50 range.
Determination of rilpivirine (TMC‐278) plasma concentrations by the conventional LC‐MS method
Authors: Shibata M , Takahashi M , Kuwahara T , Nomura T , Yokomaku Y , Sugiura W 
Published: November 2012

BackgroundRilpivirine (TMC‐278) is a second‐generation NNRTI that is highly potent against both wild‐type and drug‐resistant HIV‐1 strains. The quantification of rilpivirine in human plasma is important to support clinical studies and determine pharmacokinetic parameters of rilpivirine. Until now there has been a methodological report for the determination of rilpivirine using LC‐MS/MS. However, the MS‐MS detector needs to be delicately set and it is expensive. To bypass these difficulties, we aimed to develop more conventional procedures for determining rilpivirine plasma concentration by LC‐MS method.MethodsA Waters Alliance 2695 HPLC and a Micromass ZQ‐2000 MS, controlled with MassLynx version 4.0 software, were used for detection. Our method involves rapid liquid‐liquid drug extraction from plasma and use of gradient elution on a reversed‐phase C18 column. The mobile phase comprised 0.1 mM EDTA in 0.1% acetic acid (65%), acetonitrile (15%), and methanol (20%). Quantitative analysis detected rilpivirine at m/z 367, and the internal standard, at m/z 313, all in the form of ions.ResultsThe established LC‐MS method was validated by estimating the precision and accuracy for inter‐ and intraday analysis in the concentration range of 18–715 ng/ml. The calibration curve was linear in this range. Average accuracy ranged from 100.0 to 100.6%. Relative standard deviations of both inter‐ and intraday assays were less than 3.3%. Recovery of rilpivirine was more than 82.0%.ConclusionsOur newly developed LC‐MS method achieves the same level of reproducibility and accuracy as the LC‐MS/MS method. Our method provides a conventional, accurate and precise way to determine rilpivirine in human plasma. This method can be used in routine clinical application for HIV‐1 infected patients, and permits management of drug interactions and toxicity for rilpivirine.
Potential drug‐drug interactions in HIV‐infected children on highly active antiretroviral therapy in Lagos, Nigeria
Authors: Oshikoya K , Lawal S , Oreagba I , Ezeaka C , Temiye E , Adeyemo T 
Published: November 2012

ObjectiveMultiple therapies are common in HIV‐infected children on highly active antiretroviral therapy (HAART). These children are at risk for clinically significant drug interactions (CSDIs). While CSDIs are under‐recognized in developed countries, data are lacking for developing African countries. We aimed to investigate the prevalence of CSDIs between antiretrovirals (ARVs) and co‐prescribed drugs for children attending a large HIV clinic in Lagos, Nigeria.MethodsWe retrospectively assessed the risk for clinically significant drug interactions in children on HAART. Of the 417 patients enrolled for treatment, 80 were eligible for inclusion. We defined CSDIs as ‘major’ (capable of causing severe or permanent damage, contraindicated, avoid or not recommended by the manufacturer, or requiring dose modification) or ‘moderate’ (manufacturers advise caution, or close monitoring, or capable of causing clinical deterioration).ResultsA total of 60 (75%) patients were at risk for a CSDI resulting in major interactions in 13 (16.3%) patients and could potentially lower the plasma concentration of antiretroviral drugs in 9 (15%) patients. Major interactions most frequently involved rifampicin in 9 (11.3%) patients and artemisinin combination therapies (ACTs) in 8 (10%) patients whereas moderate interactions frequently involved ACTs (48.8%), fluconazole (36.3%), and rifampicin (11.3%). Age (p = 0.392), gender (p = 0.813), and moderate (p = 0.692) or severe (p = 0.788) malnourished state of the children were not associated with risk for CSDIs.ConclusionsThree‐quarters of children receiving ARV drugs are at risk for CSDIs. Strategies are needed to be put in place to prevent important drug interactions and to manage unavoidable interactions.
Cost‐efficacy analysis of darunavir/r monotherapy in clinical practice
Authors: Pérez Elías M , Martinez‐Colubi M , Sanz J , Gomez C , Sepulveda M , Estrada V , Moreno A , Muriel A , Dronda F , Del Palacio M , Casado J , Gómez‐Ayerbe C , Rodriguez Sagrado M , Moreno S 
Published: November 2012

Purpose of the studyTo evaluate the economic impact of a swiching strategy to DRV/r mx in clinical practice using Spanish prices.MethodsMulticenter retrospective study of four tertiary hospitals in Spain. The analysis includes 147 patients switching to DRV/r mx mainly due to toxicity or simplification from March 2009 to June 2011. The Spanish costs (ex‐factory price+VAT) per patient with HIV RNA<50 copies/ml were calculated, accounting for additional/ switch antiretroviral taken after initial treatment failure and management of adverse events. Cost of adverse events were based on a Spanish publication [1] (updated by the inflation rate until april 2012) The horizon of the analysis was of 48 weeks.Summary of resultsBaseline characteristics were: women (30.6%), median age (49 yr), IDU (45%), AIDS stage (32%), HCV coinfected (48%, 40% with advanced fibrosis), length of HIV‐RNA<1.7 before DRV/rtv mtx 67.6. Most frequent reasons for switching to DRVr mx were toxicity (62.6%) and simplification (23.8%). If a hospital with 600 patients in ART treatment, switched from 10% to 20% of its patients to DRV/r mx, there is a potential to save up to 448,000€/year.ConclusionsSwitching to DRV/r mx is a cost‐effective strategy that allows more patients to be treated for a fixed budget. Higher cost saving is expected when toxicity is the reason for switching.

Baseline HAART
DRV/r Monotherapy
Difference

Mean cost per patient (ARV)
8.471€
5.773€ ‐
2.698€

Mean cost per patient (AEs)
70€
5€ ‐
65€

Total Mean cost per patient*
8.541€
5.778€ ‐
2.762€

Number treated for 1,000,000
117
173
56

% HIV RNA<50 copies/ml (OTT)
93,10%
93,10%
0,00%

Cost per success
9.174€
6.207€ ‐
2.967€

Number of success
109
161
52

ICER (Incremental Cost Efficacy Ratio)

Dominated

Includes ARV and AEs treatment costs.

48 Weeks Cost‐Efficacy analysis: Simplification strategy to DRV/r monotherapy

% of patients that Switches to DRV/r monotherapy

10%
15%
20%

Number of patients in ARV treatment
600
600
600

Number of patients in DRV/r Monotherapy
60
90
120

Before Switch: 1 year cost
512.441€
768.662€
1.024.883€

After Switch: 1 year cost
346.698€
520.048€
693.397€

Total Cost Saving
165.743€
248.614€
331.486€

Hospital Budget Impact Analysis: assuming that 10%–20% of 600 patients in ARV treatment simplifies to DRV/r monotherapy