Initiation site of Ca(2+) entry evoked by endoplasmic reticulum Ca(2+) depletion in mouse parotid and pancreatic acinar cells

Purpose

In non-excitable cells, which include parotid and pancreatic acinar cells, Ca²⁺ entry is triggered via a mechanism known as capacitative Ca²⁺ entry, or store-operated Ca²⁺ entry. This process is initiated by the perception of the filling state of endoplasmic reticulum (ER) and the depletion of internal Ca²⁺ stores, which acts as an important factor triggering Ca²⁺ entry. However, both the mechanism of store-mediated Ca²⁺ entry and the molecular identity of store-operated Ca²⁺ channel (SOCC) remain uncertain.

Materials and Methods

In the present study we investigated the Ca²⁺ entry initiation site evoked by depletion of ER to identify the localization of SOCC in mouse parotid and pancreatic acinar cells with microfluorometeric imaging system.

Results

Treatment with thapsigargin (Tg), an inhibitor of sarco/ endoplasmic reticulum Ca²⁺-ATPase, in an extracellular Ca²⁺ free state, and subsequent exposure to a high external calcium state evoked Ca²⁺ entry, while treatment with lanthanum, a non-specific blocker of plasma Ca²⁺ channel, completely blocked Tg-induced Ca²⁺ entry. Microfluorometric imaging showed that Tg-induced Ca²⁺ entry started at a basal membrane, not a apical membrane.

Conclusion

These results suggest that Ca²⁺ entry by depletion of the ER initiates at the basal pole in polarized exocrine cells and may help to characterize the nature of SOCC.     

Ionic mechanisms of desflurane on prolongation of action potential duration in rat ventricular myocytes

Purpose

Despite the fact that desflurane prolongs the QTC interval in humans, little is known about the mechanisms that underlie these actions. We investigated the effects of desflurane on action potential (AP) duration and underlying electrophysiological mechanisms in rat ventricular myocytes.

Materials and Methods

Rat ventricular myocytes were enzymatically isolated and studied at room temperature. AP was measured using a current clamp technique. The effects of 6% (0.78 mM) and 12% (1.23 mM) desflurane on transient outward K⁺ current (I_to), sustained outward current (I_sus), inward rectifier K⁺ current (I_KI), and L-type Ca²⁺ current were determined using a whole cell voltage clamp.

Results

Desflurane prolonged AP duration, while the amplitude and resting membrane potential remained unchanged. Desflurane at 0.78 mM and 1.23 mM significantly reduced the peak I_to by 20±8% and 32±7%, respectively, at +60 mV. Desflurane (1.23 mM) shifted the steady-state inactivation curve in a hyperpolarizing direction and accelerated inactivation of the current. While desflurane (1.23 mM) had no effects on I_sus and I_KI, it reduced the L-type Ca²⁺ current by 40±6% (p<0.05).

Conclusion

Clinically relevant concentrations of desflurane appear to prolong AP duration by suppressing I_to in rat ventricular myocytes.     

Cardioprotection Via Modulation of Calcium Homeostasis by Thiopental in Hypoxia-Reoxygenated Neonatal Rat Cardiomyocytes

Purpose

Ca²⁺ homeostasis plays an important role in myocardial cell injury induced by hypoxia-reoxygenation, and prevention of intracellular Ca²⁺ overload is key to cardioprotection. Even though thiopental is a frequently used anesthetic agent, little is known about its cardioprotective effects, particulary in association with Ca²⁺ homeostasis. We investigated whether thiopental protects cardiomyocytes against hypoxia-reoxygenation injury by regulating Ca²⁺ homeostasis.

Materials and Methods

Neonatal rat cardiomyocytes were isolated. Cardiomyocytes were exposed to different concentrations of thiopental and immediately replaced in the hypoxic chamber to maintain hypoxia. After 1 hour of exposure, a culture dish was transferred to the CO₂ incubator and cells were incubated at 37℃ for 5 hours. At the end of the experiments, the authors assessed cell protection using immunoblot analysis and caspase activity. The mRNA of genes involved in Ca²⁺ homeostasis, mitochondrial membrane potential, and cellular Ca²⁺ levels were examined.

Results

In thiopental-treated cardiomyocytes, there was a decrease in expression of the proapoptotic protein Bax, caspase-3 activation, and intracellular Ca²⁺ content. In addition, both enhancement of anti-apoptotic protein Bcl-2 and activation of Erk concerned with survival were shown. Furthermore, thiopental attenuated alterations of genes involving Ca²⁺ regulation and significantly modulated abnormal changes of NCX and SERCA2a genes in hypoxia-reoxygenated neonatal cardiomyocytes. Thiopental suppressed disruption of mitochondrial membrane potential (ΔΨ_m) induced by hypoxia-reoxygenation.

Conclusion

Thiopental is likely to modulate expression of genes that regulate Ca²⁺ homeostasis, which reduces apoptotic cell death and results in cardioprotection.     

Facilitation of Ca2+-activated K+ channels (IKCa1) by mibefradil in B lymphocytes

K⁺ channels play critical roles in the proliferation and activation of lymphocytes. Mouse B cells express large-conductance background K⁺ channel (LK_bg) in addition to the voltage-gated K⁺ channel (Kv) and Ca²⁺-activated K⁺ channel current (IKCa1). Mibefradil, a blocker of T-type Ca²⁺ channels, has been reported to affect the proliferation of immune cells. In this study, we investigated the effects of mibefradil on the membrane potential and ion channels in murine B cell lines, WEHI-231 and Bal-17. In the whole-cell patch clamp experiments, mibefradil blocked Kv and LK_bg current with half inhibitory concentration (IC₅₀), 1.9 and 2.3 μM, respectively. Interestingly, IKCa1 current was increased by mibefradil. In the inside-out patch clamp study with cloned murine IKCa1 (mIKCa1) in HEK-293, mibefradil increased both Ca²⁺ sensitivity and maximum activity of mIKCa1. At high concentrations (>10 μM), mibefradil inhibited mIKCa1 in a voltage-dependent manner. Application of anti-IgM antibody to stimulate B cell receptors (BCR-ligation) induced transient hyperpolarization of Bal-17 and WEHI-231 cells, which became persistent with 1 μM mibefradil. The hyperpolarizing response was abolished by charybdotoxin, a selective blocker for SK4/IKCa1. In summary, our study firstly reports the ion channel-activating effects of mibefradil. The selective potent activation of IKCa1 suggests that mibefradil-derived drugs might be useful in the control of cell responses related with IKCa1. HY Yoo, H Zheng, and JH Nam contributed equally to this study.     

Proliferation of human lens epithelial cells (HLE-B3) is inhibited by blocking of voltage-gated calcium channels

Calcium, as an integral part of a large number of cellular regulatory pathways, is selective in the control of specific cell functions like the start of G1 phase in cell cycle. Cell proliferation has been suggested to depend on increasing intracellular calcium levels. A major regulatory pathway for intracellular calcium is the calcium influx into the cell via voltage-gated calcium channels. T-type and L-type calcium channels are substantially present in human lens epithelial cell (hLEC), and total calcium currents are inhibited by mibefradil. Here, the hypothesis was tested if calcium influx via Ca_v channels regulates proliferation in epithelial cells. Cell proliferation was determined by cell culture assays using the L- and T-type Ca_v channel blockers mibefradil and verapamil as modulators for calcium influx. Calcium influx was investigated using the Manganese quench technique. Western blot experiments were accomplished under standard conditions using antibodies against MAPK 3. Mibefradil as well as verapamil impaired cell proliferation, but in different concentration ranges. Furthermore, the activation of MAPK 3 was reduced by both antagonists. Calcium influx was also reduced in the presence of both blockers. We conclude that the transmembrane influx of Ca²⁺ through Ca_v channels contributes to the regulation of hLEC proliferation, identifying Ca_v channel blockers as potential therapeutic substances in ocular diseases.     

Impact of immunosuppressant therapy on early recurrence of hepatocellular carcinoma after liver transplantation

Background/Aims

The most commonly used immunosuppressant therapy after liver transplantation (LT) is a combination of tacrolimus and steroid. Basiliximab induction has recently been introduced; however, the most appropriate immunosuppression for hepatocellular carcinoma (HCC) patients after LT is still debated.

Methods

Ninety-three LT recipients with HCC who took tacrolimus and steroids as major immunosuppressants were included. Induction with basiliximab was implemented in 43 patients (46.2%). Mycophenolate mofetil (MMF) was added to reduce the tacrolimus dosage (n=28, 30.1%). The 1-year tacrolimus exposure level was 7.2 ± 1.3 ng/mL (mean ± SD).

Results

The 1- and 3-year recurrence rates of HCC were 12.9% and 19.4%, respectively. Tacrolimus exposure, cumulative steroid dosages, and MMF dosages had no impact on HCC recurrence. Induction therapy with basiliximab, high alpha fetoprotein (AFP; >400 ng/mL) and protein induced by vitamin K absence/antagonist-II (PIVKA-II; >100 mAU/mL) levels, and microvascular invasion were significant risk factors for 1-year recurrence (P<0.05). High AFP and PIVKA-II levels, and positive ¹⁸fluoro-2-deoxy-d-glucose positron-emission tomography findings were significantly associated with 3-year recurrence (P<0.05).

Conclusions

Induction therapy with basiliximab, a strong immunosuppressant, may have a negative impact with respect to early HCC recurrence (i.e., within 1 year) in high-risk patients.     

A clinical,microbiological and immunological comparison between subgingival irrigation with Dentol? and chlorhexidine in advanced periodontitis

Introduction

The purpose of this study was to compare the clinical, microbiological and immunological effects of subgingival irrigation with Dentol^™ and 0.2% chlorhexidine in human advanced periodontitis.

Material and methods

Twenty cases of advanced periodontitis patients were randomly treated either with Dentol^™ (as test) or 0.2% chlorhexidine (as control) every other day for 29 days after primary scaling, root planing and oral hygiene instruction. Pocket depth (PD), bleeding on probing (BOP) and plaque index (PI) were considered as clinical parameters. Total counts of aerobic and anaerobic bacteria were measured in subgingival plaque. At the end, concentrations of TNF-α and IL-1β in gingival crevicular fluid were measured as inflammatory markers.

Results

PD and BOP decreased in both groups on the 29^th day and two weeks afterward. PI did not show any statistical difference either within or among groups (p > 0.05). Results indicate that both chlorhexidine and Dentol^™ diminished total count of subgingival anaerobic bacteria significantly (p < 0.05). Total count of subgingival aerobic bacteria increased in both groups, but the differences between the two groups were statistically significant in favour of Dentol^™ (p < 0.05). The level of IL-1β in the crevicular fluid was reduced in both groups but it was more significant in the test group (p < 0.05). The TNF-a level decreased in both groups but the statistical difference between the two groups was negligible (p > 0.05).

Conclusions

Subgingivally irrigation with Dentol^™, containing 0.02% carvacrol, every other day for 29 days is equally or more effective than 0.2% chlorhexidine in reducing clinical and immunological inflammatory indices.     

KGFR promotes Na+ channel expression in a rat acute lung injury model

Background

Binding of keratinocyte growth factor (KGF) to the KGF receptor (KGFR) plays an important role in the recovery of alveolar epithelial cells from acute lung injury (ALI).

Objectives

To evaluate the effect of gene therapy via adenovirus gene transfer of KGFR on the treatment of ALI.

Methods

Sprague-Dawley rats were divided into four groups: normal controls, injury controls, normal adenovirus transduced group and injury adenovirus transduced group. The ALI model was induced by lipopolysaccharide (LPS) injection. Recombinant adenovirus (AdEasy-KGFR) was injected via the tail vein. Expression of the sodium (Na⁺) channel in rat alveolar type II (ATII) epithelial cells was determined by PCR, immunohistochemistry and immunoelectron microscopy of rat lung tissues.

Results

Gene expression of the Na⁺ channel and KGFR in ATII cells was higher in the normal adenovirus transduced group than the three other groups; expression of these two genes in the injury adenovirus transduced group was higher than the injury control group. Na⁺ channel protein expression was lower in the injury adenovirus transduced group but higher than the injury control group.

Conclusions

KGFR over-expression induced Na channel expression could potentially be beneficial for ALI therapy.     

Bone morphogenetic protein-4 induces upregulation of Cav3.1 Ca2+ channels in HL-1 atrial myocytes

Cardiac T-type Ca²⁺ channels are reexpressed in atrial and ventricular myocytes under various pathological conditions such as post-myocardial infarction, hypertrophy, and heart failure, but relatively little is known about the mechanisms. Our previous study found that bone morphogenetic protein-4 (BMP4) was reexpressed in pathological cardiac hypertrophy models and BMP4-mediated cardiomyocyte hypertrophy. We hypothesized that BMP4 could upregulate cardiac T-type Ca²⁺ channels in HL-1 atrial myocytes. The T-type Ca²⁺ currents were recorded by using the patch-clamp technique, and the expressions of Cav3.1 and Cav3.2 were measured by real-time PCR method in HL-1 cells. BMP4 and Cav3.1 mRNA expressions increased in the left atrium from the pressure overload-induced hypertrophy of mice hearts. BMP4 treatment for 48 h induced increase of Cav3.1 but not Cav3.2 mRNA expression in HL-1 cells, and the increase was inhibited by BMP4 inhibitor noggin. Acute treatment with BMP4 did not affect T-type Ca²⁺ currents, but chronic treatment (48 h) significantly increased the amplitude of T-type Ca²⁺ currents in HL-1 cells. Chronic treatment with BMP4 induced upregulation of NADPH oxidase-4 (NOX4), increase of reactive oxygen species (ROS) level, and activation of mitogen-activated protein kinase (MAPK)-activated protein kinases c-jun N-terminal kinases (JNK) and p38. BMP4-induced upregulation of Cav3.1 mRNA was inhibited by NADPH oxidase inhibitor apocynin, the radical scavenger tempol, JNK inhibitor SP600125, and p38 inhibitor SB203580. In conclusion, BMP4 induces upregulation of Cav3.1 Ca²⁺ channels and T-type Ca²⁺ currents in HL-1 atrial myocytes through ROS/MAPK pathways.     

Lack of Association of the APOE ε4 Allele with the Risk of Obstructive Sleep Apnea: Meta-Analysis and Meta-Regression

Study Objectives:

Reports on the association of polymorphisms in the gene encoding apolipoprotein E (APOE)—a vital macromolecule in cholesterol metabolism—with obstructive sleep apnea (OSA) have provided conflicting results. Our objective was to meta-analytically synthesize the existing evidence for the association of the APOE ε4 allele with the risk of OSA.

Design:

Random effects meta-analysis and meta-regression

Setting:

Genetic epidemiological studies reporting the association of APOE ε4 allele with OSA susceptibility.

Patients or Participants:

Synthesis of APOE ε4 allele data from 6,508 subjects including 1,901 cases of OSA and 4,607 controls.

Interventions:

None

Measurements and Results:

Eight studies were included in the random effects meta-analysis; the summary effect size measured as odds ratio (OR) for association of the APOE ε4 allele with the risk of OSA was found to be 1.13 (95% confidence interval 0.86–1.47). There was a statistically significant heterogeneity (I² = 72%, P = 0.001) across study results that was not explained by the mean age, proportion of males, or the proportion possessing the APOE ε4 allele or when grouped based on the geographic location of the study.

Conclusions:

The hypothesis that the APOE ε4 allele may be causally associated with OSA cannot be supported on the basis of published literature.

Citation:

Thakre TP; Mamtani MR; Kulkarni H. Lack of association of the APOE ε4 allele with the risk of obstructive sleep apnea: meta-analysis and meta-regression. SLEEP 2009;32(11):1507-1511.     

Attenuation of Peripheral Regulatory T-Cell Suppression of Skin-Homing CD8+T Cells in Atopic Dermatitis

Purpose

Cutaneous lymphocyte-associated antigen (CLA)-expressing CD8⁺T cells have been known to play an important role in the pathogenesis of atopic dermatitis (AD). However, the mechanisms underlying the loss of self-tolerance remain unclear. Regulatory T cells (Tregs) play a key role in the development of homeostasis in the immune system. We, therefore, hypothesized that a reduced ability of Tregs to inhibit autologous CD8⁺CLA⁺T cells might be underlying mechanism in AD.

Materials and Methods

CD8⁺CLA⁺T cells and Tregs were obtained from the peripheral blood of AD patients and control volunteers. The frequencies of CD8⁺CLA⁺T cells were evaluated. The proliferative responses of CD8⁺CLA⁺T cells were assessed by flow cytometry, and the levels of transforming growth factor-β1 (TGF-β1) and interleukin-10 (IL-10) in culture supernatants were detected by enzyme-linked immunosorbent assay.

Results

Our results revealed higher frequency and increased expression of perforin and granzyme-B in peripheral CD8⁺CLA⁺T cells in AD, and lower inhibitory ability of Tregs on proliferation of CD8⁺CLA⁺T cells in AD. Meanwhile, the levels of TGF-β1 produced by Tregs were significantly lower in AD, and anti-TGF-β1 abolished such suppression.

Conclusion

The attenuated inhibitory ability of Tregs on hyper-activated autologous CD8⁺CLA⁺T cells, mediated by TGF-β1, plays an important role in the pathogenesis of AD.     

Calcium-phosphate metabolism parameters and erythrocyte Ca2+ concentration in autosomal dominant polycystic kidney disease patients with normal renal function

Introduction

The aim of this study was to assess calcium-phosphate metabolism of autosomal dominant polycystic kidney disease (ADPKD) patients with a special consideration to the following serum parameters: calcium (Ca²⁺), inorganic phosphate (Pi), parathyroid hormone (PTH) and intracellular erythrocyte calcium ([Ca²⁺]_i) concentrations.

Material and methods

The study included 49 adult ADPKD patients (19 males, 30 females) aged 36 ±11 years with normal renal function and no diagnosis of diabetes as well as 50 healthy controls (22 males, 28 females) matched for age and gender. Serum concentrations of sodium (Na⁺), potassium (K⁺) and magnesium (Mg²⁺) ions and Pi were determined with an indirect ion-selective method, while Ca2+ concentration was measured with a direct ion-selective method. The PTH was detected using a radioimmunometric method. [Ca²⁺]_i concentration was determined with the Ca²⁺ sensitive fluorescent dye Fura-2 method.

Results

In the ADPKD group, when compared to controls, the following concentrations were significantly higher: serum Ca²⁺ (1.18 ±0.06 mmol/l vs. 1.15 ±0.06 mmol/l, p = 0.0085), [Ca²⁺]_i (146.9 ±110.0 nmol/l vs. 96.5 ±52.7 nmol/l, p = 0.0075), serum Na+ (139.4 ±2.7 mmol/l vs. 138.5 ±2.1 mmol/l, p = 0.060, borderline significance), and PTH (15.5 ±6.8 pg/ml vs. 13.6 ±5.3 pg/ml, p = 0.066, borderline significance), while serum Mg²⁺ was significantly lower (0.81 ±0.09 mmol/l vs. 0.85 ±0.05 mmol/l, p = 0.021). In the ADPKD group we observed significant negative correlations of PTH with Ca²⁺ serum concentrations (Rs = –0.32, p = 0.025) and with estimated glomerular filtration rate (Rs = –0.31, p = 0.033).

Conclusions

The erythrocyte Ca²⁺ concentration is elevated in ADPKD patients with normal renal function. It may result from a dysfunction of mutated polycystins which can affect various aspects of electrolyte metabolism.     

Localization of Rod Bipolar Cells in the Mammalian Retina Using an Antibody Against the α1c L-type Ca2+ Channel

Bipolar cells transmit stimuli via graded changes in membrane potential and neurotransmitter release is modulated by Ca²⁺ influx through L-type Ca²⁺ channels. The purpose of this study was to determine whether the α₁c subunit of L-type voltage-gated Ca²⁺ channel (α₁c L-type Ca²⁺ channel) colocalizes with protein kinase C alpha (PKC-α), which labels rod bipolar cells. Retinal whole mounts and vertical sections from mouse, hamster, rabbit, and dog were immunolabeled with antibodies against PKC-α and α₁c L-type Ca²⁺ channel, using fluorescein isothiocyanate (FITC) and Cy5 as visualizing agents. PKC-α-immunoreactive cells were morphologically identical to rod bipolar cells as previously reported. Their cell bodies were located within the inner nuclear layer, dendritic processes branched into the outer plexiform layer, and axons extended into the inner plexiform layer. Immunostaining showed that α₁c L-type Ca²⁺ channel colocalized with PKC-α in rod bipolar cells. The identical expression of PKC-α and α₁c L-type Ca²⁺ channel indicates that the α₁c L-type Ca²⁺ channel has a specific role in rod bipolar cells, and the antibody against the α₁c L-type Ca²⁺ channel may be a useful marker for studying the distribution of rod bipolar cells in mouse, hamster, rabbit, and dog retinas.     

Effects of heme precursors on CYP1A2 and POR expression in the baculovirus/Spodoptera frugiperda system

Objective

CYP1A2 and NADPH-CYP450 oxidoreductase (POR) were expressed in the baculovirus/Spodoptera frugiperda (sf9) system. The aim of this study was to investigate the effects of heme precursors on the expression of CYP1A2 and POR.

Methods

The heme precursors [δ-Aminolaevulinic Acid (5-ALA), Fe³⁺ and hemin] were introduced into the system to evaluate their effects on the expression of CYP1A2, POR and their co-expression. All the proteins were identified using immunoblotting, CO-difference spectroscopy, or cytochrome c assay.

Results

In the present study, functional CYP1A2 and POR were successfully expressed in the baculovirus/sf9 system, and both of them showed high activities. Co-addition of 5-ALA and Fe³⁺ significantly improved expression of CYP1A2 by about 50% compared with the addition of 5-ALA, Fe³⁺ or hemin alone. Either co-addition of 5-ALA and Fe³⁺ or addition of 5-ALA or Fe³⁺ alone improved the POR expression level 2 fold and its activity 7-10 fold compared with control (no addition). However, unlike CYP1A2, there was no difference between the co-addition and addition of these heme precursors alone. Different ratios of BvCYP1A2 to BvPOR also affected the co-expression of CYP1A2 and POR, with a 3:1 ratio of BvCYP1A2 / BvPOR significantly increasing their co-expression. Surprisingly, the addition of 0.1 mM 5-ALA or Fe³⁺ alone, but not their co-addition, could significantly improve the CYP1A2 and POR co-expression (P < 0.05).

Conclusion

5-ALA and Fe³⁺ increased the expression of CYP1A2 and POR in a baculovirus/sf9 system, but the pattern of their expression was different between their expression alone and co-expression.     

Anti-Tumour Effects of Polysaccharides Isolated from Artemisia Annua L by Inducing Cell Apoptosis and Immunomodulatory Anti-Hepatoma Effects of Polysaccharides

Background

It is well known that various polysaccharides present anti-tumour effects by inducing cell apoptosis and immunomodulation. However, it is still unclear about the roles of polysaccharides isolated from Artemisia apiacea (HQG) to hepatoma and its underlying mechanism. The objective of the study was to examine the anti-hepatoma effects of HQG and its related mechanism.

Materials and Methods

HQG was prepared in house and the quality and purity were confirmed by infra-red spectrum and gel permeation chromatography (GPC). Tumour-bearing mice induced by injection of mouse hepatoma H22 cells were used to evaluate the tumour growth inhibition by HQG administration. Cell immunostaining, JC1 staining and flow cytometer were performed to examine the cell apoptosis, mitochondrial membrane potential change and immunomodulation in response to HQG treatment.

Results

HQG treatment inhibited hepatoma growth in tumour-bearing mice. Cell apoptosis rate of human hepatoma 7402 cells and of the cells from ascites in tumour-bearing mice was increased after HQG treatment. Mitochondrial membrane potential in human hepatoma 7402 cells was decreased after HQG treatment. CD4+ and CD8+ T-lymphocytes subpopulation was increased while the ratio of CD4+/ CD8+ decreased in tumour-bearing mice after HQG administration. IFN-γ and IL-4 secretion was increased in spleen lymphocytes in tumour-bearing mice after HQG administration.

Conclusion

The study concluded that polysaccharides isolated from Artemisia apiacea (HQG) can inhibit hepatoma cell growths by facilitating cell apoptosis and immuno-defence.     

Detection and significance of TregFoxP3+ and Th17 cells in peripheral blood of non-small cell lung cancer patients

Introduction

The aim of this study was to explore the relationships between TregFoxP3⁺ cells and Th17 cells and occurrence of lung cancer.

Material and methods

The proportions of TregFoxP3⁺ and Th17 cells, the expression of FoxP3 and RORγt mRNA, and the levels of related cell factors such as transforming growth factor-β (TGF-β), interleukin IL-17 (IL-17) and IL-23 were determined respectively by flow cytometry analysis, real-time-polymerase chain reaction (PCR), and ELISA in peripheral blood of 18 healthy people and 26 patients with non-small cell lung cancer (NSCLC).

Results

The levels of TregFoxP3⁺ and Th17, expression of FoxP3 and RORγt mRNA, and ratios of TregFoxP3⁺/Th17 and FoxP3/RORγt in peripheral blood with NSCLC were higher than those in healthy controls (p < 0.05). The proportion of Th17 cells from NSCLC patients was positively correlated with that of TregFoxP3⁺ (r = 0.81, p < 0.05). The receiver-operating characteristic (ROC) curve demonstrates that the increased level of TregFoxP3⁺/Th17 in the peripheral blood may be a useful indicator in early diagnosis of non-small cell lung carcinoma. The TregFoxP3⁺/Th17 and FoxP3/RORγt levels for patients in stage IV were higher than those of patients in stages I, II, and III (p < 0.05). The levels of TGF-β, IL-17, and IL-23 were higher in NSCLC patients than those in healthy controls.

Conclusions

The results suggest that ratios of Treg/Th17 correlate with the stage of NSCLC.     

Synergistic effect of aqueous extract of Telfaria occidentalis on the biological activities of artesunate in Plasmodium berghei infected mice

Background

Resistance to most antimalarial drugs has encouraged the use of herbal preparations along with prescribed orthodox drugs.

Objective

To investigate effect of co-administration of aqueous extract of T. occidentalis leaves; commonly used as antimalarial and haematinic agent in Nigeria and artesunate using P. berghei animal model.

Methods

In vivo curative antiplasmodial effect of T. occidentalis (200mg/kg) alone and combination with artesunate (2mg/kg) were evaluated using albino mice infected with 10⁶ parasitized erythrocytes of P. berghei intraperitoneally. The haematological parameters: haemoglobin level, red blood cells and white blood cells and packed cell volume were monitored using standard methods.

Results

Aqueous extract of T. occidentalis, artesunate and the combination gave 72.17±4.07%, 70.43± 4.27% and 85.43±3.65% reduction in parasitaemia after 48hours respectively. A significant enhancement of the PCV was obtained with the coadministration of artesunate and aqueous extract (p< 0.01). Similar trends were also observed with heamatological parameters at 72hours of administration.

Conclusion

This study revealed a synergistic effect of the co-administration on parasite clearance rate of P. berghei infection in mice, with a significant enhancement of haematological parameters within 48 hours of administration. This indicates a rapid rate of recovery from plasmodial infections with the co-administration.     

Piper sarmentosum enhances fracture healing in ovariectomized osteoporotic rats: a radiological study

INTRODUCTION:

Osteoporotic fractures are common during osteoporotic states. Piper sarmentosum extract is known to possess antioxidant and anti-inflammatory properties.

OBJECTIVES:

To observe the radiological changes in fracture calluses following administration of a Piper sarmentosum extract during an estrogen-deficient state.

METHODS:

A total of 24 female Sprague-Dawley rats (200-250 g) were randomly divided into 4 groups: (i) the sham-operated group; (ii) the ovariectomized-control group; (iii) the ovariectomized + estrogen-replacement therapy (ovariectomized-control + estrogen replacement therapy) group, which was supplemented with estrogen (100 µg/kg/day); and (iv) the ovariectomized + Piper sarmentosum (ovariectomized + Piper sarmentosum) group, which was supplemented with a water-based Piper sarmentosum extract (125 mg/kg). Six weeks after an ovariectomy, the right femora were fractured at the mid-diaphysis, and a K-wire was inserted. Each group of rats received their respective treatment for 6 weeks. Following sacrifice, the right femora were subjected to radiological assessment.

RESULTS:

The mean axial callus volume was significantly higher in the ovariectomized-control group (68.2±11.74 mm³) than in the sham-operated, estrogen-replacement-therapy and Piper sarmentosum groups (20.4±4.05, 22.4±4.14 and 17.5±3.68 mm³, respectively). The median callus scores for the sham-operated, estrogen-replacement-therapy and Piper sarmentosum groups had median (range, minimum - maximum value) as 1.0 (0 - 2), 1.0 (1 - 2) and 1.0 (1 - 2), respectively, which were significantly lower than the ovariectomized-control group score of 2.0 (2 - 3). The median fracture scores for the sham-operated, estrogen-replacement-therapy and Piper sarmentosum groups were 3.0 (3 - 4), 3.0 (2 - 3) and 3.0 (2 - 3), respectively, which were significantly higher than the ovariectomized-control group score of 2.0 (1 - 2) (p<0.05).

CONCLUSION:

The Piper sarmentosum extract improved fracture healing, as assessed by the reduced callus volumes and reduced callus scores. This extract is beneficial for fractures in osteoporotic states.     

Relationship between Preoperative 18F-Fluorodeoxyglucose Uptake and Epidermal Growth Factor Receptor Status in Primary Colorectal Cancer

Purpose

Both ¹⁸F-fluorodeoxyglucose (¹⁸F-FDG) uptake and epidermal growth factor receptor (EGFR) status are prognostic variables of colorectal cancer (CRC). The aim of this study was to investigate a possible association between ¹⁸F-FDG uptake on preoperative positron emission tomography/computed tomography (PET/CT) and EGFR status in primary CRC.

Materials and Methods

Records of 132 patients (66 men and 66 women; mean age=67.1±11.1 years) who underwent ¹⁸F-FDG PET/CT for CRC staging and subsequent bowel resection were reviewed. In primary lesions, ¹⁸F-FDG uptake was semiquantitatively evaluated in terms of maximum standardized uptake value (SUV_max), and EGFR status was determined by immunohistochemistry. Associations of clinicopathological parameters and EGFR status were analyzed by Pearson''s chi-square test, multiple logistic regression, and receiver operating characteristic curves.

Results

Eighty-six patients (65.2%) showed EGFR expression. SUV_max was significantly lower in EGFR-negative tumors than in EGFR-expressing tumors (10.0±4.2 vs. 12.1±2.1; p=0.012). It was the only significant parameter correlated with EGFR expression (odds ratio=2.457; relative risk=2.013; p=0.038). At the SUV_max threshold of 7.5, the sensitivity and specificity for predicting EGFR expression were 84.9% and 40.4%, respectively (area under the curve=0.624; p=0.019).

Conclusion

Preoperative ¹⁸F-FDG uptake is slightly correlated with EGFR status in primary CRC. Preoperative SUV_max of ¹⁸F-FDG may have a limited role in predicting EGFR expression in such tumors because of its poor specificity.