Diagnostic Accuracy of Microdialysis in Postoperative Flap Monitoring.

Postoperative monitoring plays an important role in achieving success in microvascular free tissue transfer. A systematic review was designed to evaluate the clinical outcomes of microdialysis in flap monitoring and a meta-analysis was conducted for diagnostic accuracies. The search terms "microdialysis" and "flap" were used in a PubMed and Scopus search, resulting in 60 and 78 results, respectively. Among 78 titles, 15 articles were excluded. Among 63 abstracts, 43 abstracts were excluded. From 20 full texts, 7 articles were excluded because they did not have sufficient content (ie, the statistical values in question). A systematic review was conducted of the final 13 articles. The overall sensitivity was 97.24% [95% confidence interval (CI)=93.67%-99.10%]. Eleven of the 13 studies showed 100% sensitivity and 2 studies had 2 and 3 false negative results, resulting in sensitivity values of 85.8% and 95.3%. Specificity ranged from 91.89% to 100%, and the overall value was 98.15% (95% CI=96.80%-99.04%). The positive predictive value ranged from 84.62% to 100%, with an overall value of 93.62% (95% CI=89.33%-96.26%). The negative predictive value ranged from 94.44% to 100%, with an overall value of 99.22% (95% CI=98.17%-99.67%). The overall flap success rate (survival rate) was 93.7% (786/839). The lowest flap survival rate was 86.7% and the highest was 100%. Microdialysis provides excellent diagnostic accuracy and enables the early detection of ischemia in postoperative flap monitoring. Although microdialysis is not the most popular choice among surgeons, it should be considered adjacent to conventional clinical monitoring. Cost-effectiveness, availability, and ease of application remain hurdles.

Gα12 overexpression induced by miR-16 dysregulation contributes to liver fibrosis by promoting autophagy in hepatic stellate cells.

BACKGROUND & AIMS: Hepatic stellate cells (HSCs) have a role in liver fibrosis. Guanine nucleotide-binding α-subunit 12 (Gα12) converges signals from G-protein-coupled receptors whose ligand levels are elevated in the environment during liver fibrosis; however, information is lacking on the effect of Gα12 on HSC trans-differentiation. This study investigated the expression of Gα12 in HSCs and the molecular basis of the effects of its expression on liver fibrosis. METHODS: Gα12 expression was assessed by immunostaining, and immunoblot analyses of mouse fibrotic liver tissues and primary HSCs. The role of Gα12 in liver fibrosis was estimated using a toxicant injury mouse model with Gα12 gene knockout and/or HSC-specific Gα12 delivery using lentiviral vectors, in addition to primary HSCs and LX-2 cells using microRNA (miR) inhibitors, overexpression vectors, or adenoviruses. miR-16, Gα12, and LC3 were also examined in samples from patients with fibrosis. RESULTS: Gα12 was overexpressed in activated HSCs and fibrotic liver, and was colocalised with desmin. In a carbon tetrachloride-induced fibrosis mouse model, Gα12 ablation prevented increases in fibrosis and liver injury. This effect was attenuated by HSC-specific lentiviral delivery of Gα12. Moreover, Gα12 activation promoted autophagy accompanying c-Jun N-terminal kinase-dependent ATG12-5 conjugation. In addition, miR-16 was found to be a direct inhibitor of the de novo synthesis of Gα12. Modulations of miR-16 altered autophagy in HSCs. In a fibrosis animal model or patients with severe fibrosis, miR-16 levels were lower than in their corresponding controls. Consistently, cirrhotic patient liver tissues showed Gα12 and LC3 upregulation in desmin-positive areas. CONCLUSIONS: miR-16 dysregulation in HSCs results in Gα12 overexpression, which activates HSCs by facilitating autophagy through ATG12-5 formation. This suggests that Gα12 and its regulatory molecules could serve as targets for the amelioration of liver fibrosis. LAY SUMMARY: Guanine nucleotide-binding α-subunit 12 (Gα12) is upregulated in activated hepatic stellate cells (HSCs) as a consequence of the dysregulation of a specific microRNA that is abundant in HSCs, facilitating the progression of liver fibrosis. This event is mediated by c-Jun N-terminal kinase-dependent ATG12-5 formation and the promotion of autophagy. We suggest that Gα12 and its associated regulators could serve as new targets in HSCs for the treatment of liver fibrosis.

Changes in the Expressions of Iba1 and Calcitonin Gene-Related Peptide in Adjacent Lumbar Spinal Segments after Lumbar Disc Herniation in a Rat Model.

Lumbar disc herniation is commonly encountered in clinical practice and can induce sciatica due to mechanical and/or chemical irritation and the release of proinflammatory cytokines. However, symptoms are not confined to the affected spinal cord segment. The purpose of this study was to determine whether multisegmental molecular changes exist between adjacent lumbar spinal segments using a rat model of lumbar disc herniation. Twenty-nine male Sprague-Dawley rats were randomly assigned to either a sham-operated group (n=10) or a nucleus pulposus (NP)-exposed group (n=19). Rats in the NP-exposed group were further subdivided into a significant pain subgroup (n=12) and a no significant pain subgroup (n=7) using mechanical pain thresholds determined von Frey filaments. Immunohistochemical stainings of microglia (ionized calcium-binding adapter molecule 1; Iba1), astrocytes (glial fibrillary acidic protein; GFAP), calcitonin gene-related peptide (CGRP), and transient receptor potential vanilloid 1 (TRPV1) was performed in spinal dorsal horns and dorsal root ganglions (DRGs) at 10 days after surgery. It was found immunoreactivity for Iba1-positive microglia was higher in the L5 (P=0.004) dorsal horn and in the ipsilateral L4 (P=0.009), L6 (P=0.002), and S1 (P=0.002) dorsal horns in the NP-exposed group than in the sham-operated group. The expression of CGRP was also significantly higher in ipsilateral L3, L4, L6, and S1 segments and in L5 DRGs at 10 days after surgery in the NP-exposed group than in the sham-operated group (P<0.001). Our results indicate that lumbar disc herniation upregulates microglial activity and CGRP expression in many adjacent and ipsilateral lumbar spinal segments.

Skin tension related to tension reduction sutures.

The aim of this study was to compare the skin tension of several fascial/subcutaneous tensile reduction sutures. Six upper limbs and 8 lower limbs of 4 fresh cadavers were used. At the deltoid area (10 cm below the palpable acromion) and lateral thigh (midpoint from the palpable greater trochanter to the lateral border of the patella), and within a 3 × 6-cm fusiform area of skin, subcutaneous tissue defects were created. At the midpoint of the defect, a no. 5 silk suture was passed through the dermis at a 5-mm margin of the defect, and the defect was approximated. The initial tension to approximate the margins was measured using a tensiometer.The tension needed to approximate skin without any tension reduction suture (S) was 6.5 ± 4.6 N (Newton). The tensions needed to approximate superficial fascia (SF) and deep fascia (DF) were 7.8 ± 3.4 N and 10.3 ± 5.1 N, respectively. The tension needed to approximate the skin after approximating the SF was 4.1 ± 3.4 N. The tension needed to approximate the skin after approximating the DF was 4.9 ± 4.0 N. The tension reduction effect of approximating the SF was 38.8 ± 16.4% (2.4 ± 1.5 N, P = 0.000 [ANOVA, Scheffé]). The tension reduction effect of approximating the DF was 25.2% ± 21.9% (1.5 ± 1.4 N, P = 0.001 [ANOVA, Scheffé]). The reason for this is thought to be that the SF is located closely to the skin unlike the DF. The results of this study might be a basis for tension reduction sutures.

Ideal or Young-Looking Brow Height and Arch Shape Preferred by Koreans.

The aim of this study is to see which brow height and arch shape is preferred as ideal or young-looking by Koreans. A survey was conducted between June and Dec 2014 on 186 women who visited the brow bar ("Benefit" of Incheon city). They were asked to choose which they believed ideal and youngest amongst the 3 brow archetypes according to their height and 4 types of modification of Anastasia (rotation of medial and lateral arms), which was illustrated. Approximately half (52.5%) of the respondents answered that their brow matches them very well or well. Most (81.2%) believed there might be a method to yield an ideal brow archetype and almost all (97.3%) would change the brow shape if the expert advised. The most preferred ideal brow height was of a middle height (63.2%, the distance from the lateral canthus to the lateral end of eyebrow, which was 2/3 of the eye width). The most preferred ideal brow arch shape was the arched type (57.6% arches on a line drawn from the center of the nose through the center of the pupil). The most preferred young-looking brow height was of an upper height (46.2%, the distance from the lateral canthus to the lateral end of eyebrow was 3/4 of the eye width) followed by a middle height (39.7%). The most preferred young-looking brow arch shape was the head-up position (53.3%, medial arm of the brow was rotated upward to the horizontal plane). The result of this study might be useful in facial rejuvenation surgeries as well as in brow esthetics or tattooing of the eyebrows.

Discovery of an integrative network of microRNAs and transcriptomics changes for acute kidney injury.

The contribution of miRNA to the pathogenesis of acute kidney injury (AKI) is not well understood. Here we evaluated an integrative network of miRNAs and mRNA data to discover a possible master regulator of AKI. Microarray analyses of the kidneys of mice treated with cisplatin were used to extract putative miRNAs that cause renal injury. Of them, miR-122 was mostly downregulated by cisplatin, whereas miR-34a was upregulated. A network integrating dysregulated miRNAs and altered mRNA expression along with target prediction enabled us to identify Foxo3 as a core protein to activate p53. The miR-122 inhibited Foxo3 translation as assessed using an miR mimic, an inhibitor, and a Foxo3 3'-UTR reporter. In a mouse model, Foxo3 levels paralleled the degree of tubular injury. The role of decreased miR-122 in inducing Foxo3 during AKI was strengthened by the ability of the miR-122 mimic or inhibitor to replicate results. Increase in miR-34a also promoted the acetylation of Foxo3 by repressing Sirt1. Consistently, cisplatin facilitated the binding of Foxo3 and p53 for activation, which depended not only on decreased miR-122 but also on increased miR-34a. Other nephrotoxicants had similar effects. Among targets of p53, Phlda3 was robustly induced by cisplatin, causing tubular injury. Consistently, treatment with miR mimics and/or inhibitors, or with Foxo3 and Phlda3 siRNAs, modulated apoptosis. Thus, our results uncovered an miR integrative network regulating toxicant-induced AKI and identified Foxo3 as a bridge molecule to the p53 pathway.

Farnesoid X receptor protects hepatocytes from injury by repressing miR-199a-3p, which increases levels of LKB1.

BACKGROUND & AIMS: Hepatocyte injury occurs during liver fibrogenesis. MicroRNAs (miRNA) regulate some of these processes, and some are regulated by the farnesoid X receptor (FXR). We investigated the effect of repression of specific miRNAs by FXR in hepatocyte injury using fibrotic liver tissue from patients and hepatocytes. METHODS: We used immunohistochemistry or real-time polymerase chain reaction to analyze proteins and miRNAs in human and mouse liver samples. HepG2 cells were transfected with pre-miRNA, antisense oligonucleotides, small interfering RNAs, the 3'-untranslated region of liver kinase B1 (LKB1) (STK11), or constructs for overexpression, and analyzed. RESULTS: Liver tissue from patients with severe fibrosis had lower levels of FXR and greater amounts of hepatocyte death than samples from patients with mild disease. Levels of several miRNAs changed when FXR expression was disrupted in the liver; one of these, miR-199a-3p, was significantly up-regulated in patients with severe fibrosis. Activation of FXR by its ligand reduced the level of miR-199a-3p in HepG2 cells. LKB1 messenger RNA was identified as a target of miR-199a-3p, and its expression was reduced in human fibrotic liver tissue. Overexpression of FXR or incubation of cultured hepatocytes with the FXR ligand up-regulated LKB1; LKB1 was not induced in cells transfected with miR-199a-3p. Incubation of HepG2 cells with FXR ligand, or injection of the ligand into mice, protected hepatocytes from injury and increased levels of LKB1; levels of miR-199a-3p were reduced compared with cells that were not incubated with the FXR ligand. Activation of FXR reduced mitochondrial dysfunction and oxidative stress and increased hepatocyte survival. CONCLUSIONS: In hepatocytes, FXR represses production of miR-199a-3p. In fibrotic livers of humans and mice, FXR expression is reduced, increasing levels of miR-199a-3p, which reduces levels of LKB1. FXR therefore protects hepatocytes from injury by repressing miR-199a-3p and thereby increasing levels of LKB1.

Attenuation of spinal cord injury-induced astroglial and microglial activation by repetitive transcranial magnetic stimulation in rats.

Spinal cord injury (SCI) causes not only loss of sensory and motor function below the level of injury but also chronic pain, which is difficult and challenging of the treatment. Repetitive transcranial magnetic stimulation (rTMS) to the motor cortex, of non-invasive therapeutic methods, has the motor and sensory consequences and modulates pain in SCI-patients. In the present study, we studied the effectiveness of rTMS and the relationship between the modulation of pain and the changes of neuroglial expression in the spinal cord using a rat SCI-induced pain model. Elevated expressions of Iba1 and GFAP, specific microglial and astrocyte markers, was respectively observed in dorsal and ventral horns at the L4 and L5 levels in SCI rats. But in SCI rats treated with 25 Hz rTMS for 8 weeks, these expressions were significantly reduced by about 30%. Our finding suggests that this attenuation of activation by rTMS is related to pain modulation after SCI. Therefore, rTMS might provide an alternative means of attenuating neuropathic pain below the level of SCI.

Global plastic surgeons images depicted in motion pictures.

Motion pictures are made to entertain and enlighten people, but they are viewed differently by different people. What one considers to be a tearjerker may induce giggles in another. We have gained added interest in this because our professional pictures contain plastic surgery in their venue. We have recently reviewed 21 motion pictures that were made from 1928 to 2006 and that includes plastic surgical procedures in their content. As a habit, we tried to analyze them from a surgical point of view. About one third (35.7%) of the patients were criminals, whereas 14.3% of them were spies. One third of the procedures were done by illegitimate "surgeons," whereas a quarter of the procedures (25%) were performed by renowned surgeons. Surgeons who were in love with the patients did the rest (25%) of the operations. The complication rate was 14.3%; the surgery was successful in 85.7% of cases, but were the patients happy with the results? This was not the case in the movies. Only 7.7% were happy; 14.5 % of them were eminently unhappy. Why the discrepancy? It is difficult to analyze the minds of the people in the film, but considering that the majority of the characters in the films were rather unsavory, one may deduce that a crooked mind functions differently. Motion pictures have advanced greatly in the past several decades with the advent of improved mechanical and electronic devices, and plastic surgery as also advanced in tandem. This surgical field has become a common procedure in our daily life. It is readily available and mostly painless. However, the public sees it in only one way, that is, that the performing physicians are highly compensated. Very few consider the efforts and the suffering that accompanies each and every surgical procedure as it is performed. Perhaps, it is too much to hope for a day that will come when we will see a film that portrays the mental anguish that accompanies each and every procedure the plastic surgeon makes.

Application of stacked autoencoder for identification of bone fracture.

This study presents a stacked autoencoder (SAE)-based assessment method which is one of the unsupervised learning schemes for the investigation of bone fracture. Relatively accurate health monitoring of bone fracture requires considering physical interactions among tissue, muscle, wave propagation and boundary conditions inside the human body. Furthermore, the investigation of fracture, crack and healing process without state-of-the-art medical devices such as CT, X-ray and MRI systems is challenging. To address these issues, this study presents the SAE method that incorporates bilateral symmetry of the human legs and low-frequency transverse vibration. To verify the presented method, several examples are employed with plastic pipes, cadaver legs and human legs. Virtual spectrograms, created by applying a short-time Fourier transform to the differences in vibration responses, are employed for image-based training in SAE. The virtual spectrograms are then classified and the fine-tuning is also carried out to increase the accuracy. Moreover, a confusion matrix is employed to evaluate classification accuracy and training validity.

Pulsed radiofrequency application reduced mechanical hypersensitivity and microglial expression in neuropathic pain model.

OBJECTIVE: Pulsed radiofrequency (PRF) procedure has been used in clinical practice for the treatment of chronic neuropathic pain conditions without neuronal damage. The purpose of this study was to investigate the changes in pain response and glial expression after the application of PRF on a dorsal root ganglion (DRG) in a neuropathic pain model. DESIGN: A neuropathic pain model (14 female Sprague-Dawley [SD] rats; 200-250 g) was made by a unilateral L5 spinal nerve ligation (SNL) and transection on the distal side of the ligation. The development of mechanical and cold hypersensitivity on the hindpaw was established postoperative day 9 (POD 9). The rats were then randomly assigned to the PRF (+) and the PRF (-) groups. Furthermore, PRF (2 bursts/s, duration = 20 milliseconds, output voltage = 45 V) was applied on the ipsilateral DRG for 180 seconds, with a maximum temperature of 42°C, at POD 10. Pain behaviors were tested throughout the 12 days after PRF. We also examined the changes of the spinal glial expression by immunohistochemistry. RESULTS: Significant reduction of mechanical hypersensitivity in the PRF (+) group was observed from day 1 after a single PRF procedure and was maintained throughout the following 12 days. Immunoreactivity for OX42 in the ipsilateral dorsal horn also decreased compared with that of the PRF (-) group. However, cold hypersensitivity and glial fibrillary acidic protein (GFAP) immunoreactivity in the dorsal horn was not affected by a PRF procedure. CONCLUSIONS: Our result demonstrated that the mechanical hypersensitivity, induced by L5 SNL, was attenuated by a PRF procedure on the ipsilateral DRG. This analgesic effect may be associated with an attenuation of the microglial activation in the dorsal horn.

Effects of statin and deferoxamine administration on neurological outcomes in a rat model of intracerebral hemorrhage.

Deferoxamine (DFX), a potent iron-chelating agent, reduces brain edema and neuronal cell injury that develop due to the hemolysis cascade. Statins have neuroprotective effects via anti-inflammatory action and increment of cerebral blood flow after intracerebral hemorrhage (ICH). The purpose of this study was to identify the effects of combined DFX and statins treatment in an experimental ICH rat model. The treatments were: intraperitoneal (i.p.) injection of DFX (group I), combined treatment of i.p. DFX and oral statins (group II), statins only (group III) and treatment with vehicle (group IV). Induction of ICH was performed with injection of bacterial collagenase type IV into the left striatum. After removal of the brain, hematoma volume, water content and brain atrophy were measured. Immunohistochemistry in the perihematomal region was performed for identification of microglial infiltration, astrocyte expression and apoptotic cell presence. Statistical analysis was performed using the non-parametric Kruskal-Wallis test and significance was evaluated when the p value was less than 0.05. According to behavioral tests, significant differences among treatment groups were noted 4 weeks after ICH induction (p < 0.05). However, there were no significant differences among treatment groups in hematoma volume, brain water content or brain atrophy. In the perihematomal area, the activated microglial cells were reduced in the combined treatment group. Among the four groups, a significant difference in immunohistochemical staining was identified (p < 0.05). These results suggest that combined treatment with DFX and statins improves neurologic outcomes after ICH through reduction of microglial infiltration, apoptosis, inflammation and brain edema.

E-cadherin antagonizes transforming growth factor ß1 gene induction in hepatic stellate cells by inhibiting RhoA-dependent Smad3 phosphorylation.

UNLABELLED: Cadherins mediate cell-cell adhesion and catenin (ctn)-related signaling pathways. Liver fibrosis is accompanied by the loss of E-cadherin (ECAD), which promotes the process of epithelial-mesenchymal transition. Currently, no information is available about the inhibitory role of ECAD in hepatic stellate cell activation. Because of ECAD's potential for inhibiting the induction of transforming growth factor ß1 (TGFß1), we investigated whether ECAD overexpression prevents TGFß1 gene induction; we also examined what the molecular basis could be. Forced expression of ECAD decreased α-smooth muscle actin and vimentin levels and caused decreases in the constitutive and inducible expression of the TGFß1 gene and its downstream genes. ECAD overexpression decreased Smad3 phosphorylation, weakly decreased Smad2 phosphorylation, and thus inhibited Smad reporter activity induced by either treatment with TGFß1 or Smad3 overexpression. Overexpression of a dominant negative mutant of ras homolog gene family A (RhoA) diminished the ability of TGFß1 to elicit its own gene induction. Consistently, transfection with a constitutively active mutant of RhoA reversed the inhibition of TGFß1-inducible or Smad3-inducible reporter activity by ECAD. Studies using the mutant constructs of ECAD revealed that the p120-ctn binding domain of ECAD was responsible for TGFß1 repression. Consistently, ECAD was capable of binding p120-ctn, which recruited RhoA; this prevented TGFß1 from increasing RhoA-mediated Smad3 phosphorylation. In the liver samples of patients with mild or severe fibrosis, ECAD expression reciprocally correlated with the severity of fibrosis. CONCLUSION: Our results demonstrate that ECAD inhibits Smad3/2 phosphorylation by recruiting RhoA to p120-ctn at the p120-ctn binding domain, whereas the loss of ECAD due to cadherin switching promotes the up-regulation of TGFß1 and its target genes, and facilitates liver fibrosis.

Investigation of bone fracture diagnosis system using transverse vibration response.

In this study, a new diagnostic system is developed to easily identify bone fractures in non-medical environments. It is difficult to determine the extent of cracks, fractures, and the healing process inside humans owing to the differences among people and limitations of state-of-the-art medical devices. Thus, various medical techniques, such as X-ray, computed tomography, or fork tuning systems have been developed, and more advanced technologies are emerging in the medical engineering field. In hazardous circumstances, medical devices to detect bone fracture are not available or cannot be easily applied. Thus, there is a need for the rapid detection of bone fractures without medical devices. To this end, this study analyzes the transverse vibration responses of bones because bone fractures cause different mechanical vibration reactions. By comparing the transverse vibration responses of both healthy and fractured bones, the modal assurance criterion can be calculated and applied to detect the existence of bone fractures. The transverse vibration responses at low and high frequencies are different and exhibit different modal assurance criteria depending on whether or not they are abnormal. Then, the virtual spectrogram of the differences between the signals from non-fractured and fractured bones is calculated. With the help of the present criterion with transverse vibration data, this difference can be analyzed quantitatively and effectively. To validate the proposed system, experiments with artificial specimens, animal legs, and a cadaver are performed.

Experimental study of the effect of the boundary conditions of fractured bone.

Reliable fracture diagnosis monitoring and analyzing low-frequency transverse vibration data can be achieved through an in-depth understanding of the physical interactions between wave propagation and boundary conditions. The present study aims to investigate the effects of the boundary conditions on the low-frequency structural vibrations of bones. Time-frequency domain analysis of transverse vibration signals depending on the boundary conditions of bones is analyzed and investigated. These studies reveal that the responses of fractured or non-fractured bones are different and influenced by the displacement and force boundary conditions. These relationships can be considered in the development of a smart fracture diagnosis system considering the posture and boundary condition. To validate the present observations, the experiments with artificial specimens and cadaver are carried.

Membrane-delimited coupling of TRPV1 and mGluR5 on presynaptic terminals of nociceptive neurons.

Transient receptor potential vanilloid subtype 1 (TRPV1) and metabotropic glutamate receptor 5 (mGluR5) located on peripheral sensory terminals have been shown to play critical roles in the transduction and modulation of pain sensation. To date, however, very little is known regarding the significance of functional expression of mGluR5 and TRPV1 on the central terminals of sensory neurons in the dorsal horn of the spinal cord. Here we show that TRPV1 on central presynaptic terminals is coupled to mGluR5 in a membrane-delimited manner, thereby contributing to the modulation of nociceptive synaptic transmission in the substantia gelatinosa neurons of the spinal cord. Further, our results demonstrate that TRPV1 is involved in the pain behaviors induced by spinal mGluR5 activation, and diacylglycerol produced by the activation of mGluR5 mediates functional coupling of mGluR5 and TRPV1 on the presynaptic terminals. Thus, mGluR5-TRPV1 coupling on the central presynaptic terminals of nociceptive neurons may be an important mechanism underlying central sensitization under pathological pain conditions.

Chemokines released from astrocytes promote chemokine receptor 5-mediated neuronal cell differentiation.

Astrocytes are one of major glial cell types in the central nervous system (CNS), and can support many functions of neuronal cells. In the present study, we demonstrated that the differentiation of rat embryonic neuronal cells was promoted by treatment with astrocyte and microglia-conditioned medium. Cytokine assays identified that the IL-4, MIP-1, KC, and RANTES as were released from astrocyte, and these chemokines promote differentiation of rat embryonic neuronal cells. However, chemokine-promoted neuronal cell differentiation was suppressed by antibodies of these chemokines and their receptor (CCR5). CCR5 and neuronal cell differentiation marker proteins were found to be colocalized, and their expressions were enhanced by chemokines. Furthermore, the differentiation of neuronal cells from CCR5 knock-out mice and of neuronal cells from mice knocked down with the CCR5 siRNA were significantly reduced and delayed. Bradykinin elevated calcium influx in the embryonic neuronal cells. These data suggest that specific chemokines derived from astrocytes may significantly have influence on the CCR5-mediated differentiation of embryonic neuronal cells.

Proinflammatory cytokine and nitric oxide production by human macrophages stimulated with Trichomonas vaginalis.

Trichomonas vaginalis commonly causes vaginitis and perhaps cervicitis in women and urethritis in men and women. Macrophages are important immune cells in response to T. vaginalis infection. In this study, we investigated whether human macrophages could be involved in inflammation induced by T. vaginalis. Human monocyte-derived macrophages (HMDM) were co-cultured with T. vaginalis. Live, opsonized-live trichomonads, and T. vaginalis lysates increased proinflammatory cytokines, such as TNF-alpha, IL-1beta, and IL-6 by HMDM. The involvement of nuclear factor (NF)-kappaB signaling pathway in cytokine production induced by T. vaginalis was confirmed by phosphorylation and nuclear translocation of p65 NF-kappaB. In addition, stimulation with live T. vaginalis induced marked augmentation of nitric oxide (NO) production and expression of inducible NO synthase (iNOS) levels in HMDM. However, trichomonad-induced NF-kappaB activation and TNF-alpha production in macrophages were significantly inhibited by inhibition of iNOS levels with L-NMMA (NO synthase inhibitor). Moreover, pretreatment with NF-kappaB inhibitors (PDTC or Bay11-7082) caused human macrophages to produce less TNF-alpha. These results suggest that T. vaginalis stimulates human macrophages to produce proinflammatory cytokines, such as IL-1, IL-6, and TNF-alpha, and NO. In particular, we showed that T. vaginalis induced TNF-alpha production in macrophages through NO-dependent activation of NF-kappaB, which might be closely involved in inflammation caused by T. vaginalis.