Agonist of growth hormone-releasing hormone enhances retinal ganglion cell protection induced by macrophages after optic nerve injury.

Optic neuropathies are leading causes of irreversible visual impairment and blindness, currently affecting more than 100 million people worldwide. Glaucoma is a group of optic neuropathies attributed to progressive degeneration of retinal ganglion cells (RGCs). We have previously demonstrated an increase in survival of RGCs by the activation of macrophages, whereas the inhibition of macrophages was involved in the alleviation on endotoxin-induced inflammation by antagonist of growth hormone-releasing hormone (GHRH). Herein, we hypothesized that GHRH receptor (GHRH-R) signaling could be involved in the survival of RGCs mediated by inflammation. We found the expression of GHRH-R in RGCs of adult rat retina. After optic nerve crush, subcutaneous application of GHRH agonist MR-409 or antagonist MIA-602 promoted the survival of RGCs. Both the GHRH agonist and antagonist increased the phosphorylation of Akt in the retina, but only agonist MR-409 promoted microglia activation in the retina. The antagonist MIA-602 reduced significantly the expression of inflammation-related genes Il1b, Il6, and Tnf Moreover, agonist MR-409 further enhanced the promotion of RGC survival by lens injury or zymosan-induced macrophage activation, whereas antagonist MIA-602 attenuated the enhancement in RGC survival. Our findings reveal the protective effect of agonistic analogs of GHRH on RGCs in rats after optic nerve injury and its additive effect to macrophage activation, indicating a therapeutic potential of GHRH agonists for the protection of RGCs against optic neuropathies especially in glaucoma.

Goats with low levels of AAV antibody may serve as candidates for large animal gene therapy.

AAV vector-mediated gene therapy has been proposed as a feasible strategy for several eye diseases. However, AAV antibodies in the serum prior to treatment hinder the transduction efficiency and thus the therapeutic effect. Therefore, it is necessary to evaluate AAV antibodies in the serum before gene therapy. As large animals, goats are more closely related to humans than rodents and more economically available than nonhuman primates. Here, we first evaluated the AAV2 antibody serum level in rhesus monkeys before AAV injection. Then, we optimized a cell-based neutralizing antibody assay for detecting AAV antibodies in the serum of Saanen goats and evaluated the consistency of the cell-based neutralizing antibody assay and ELISA for goat serum antibody evaluation. The cell-based neutralizing antibody assay showed that the percentage of macaques with low antibody levels was 42.86%; however, there were no macaques with low antibody levels when the serum was evaluated by ELISA. The proportion of goats with low antibody levels was 56.67% according to the neutralizing antibody assay and 33. 33% according to the ELISA, and McNemar's test showed that the results of the two assays were not significantly different (P = 0.754), but that their consistency is poor (Kappa = 0.286, P = 0.114). Moreover, longitudinal evaluation of serum antibodies before and after intravitreal injection of AAV2 in goats revealed that the level of AAV antibodies increased and transduction inhibition subsequently increased, as reported in humans, indicating that transduction inhibition should be taken into account at different stages of gene therapy. In summary, starting with an evaluation of monkey serum antibodies, we optimized a detection method of goat serum antibodies, providing an alternative large animal model for gene therapy, and our serum antibody measurement method may be applied to other large animals.

Longitudinal evaluation of immediate inflammatory responses after intravitreal AAV2 injection in rats by optical coherence tomography.

Gene therapy has been proposed as a feasible strategy for RGC survival and optic nerve regeneration. Some preclinical and clinical studies revealed intraocular inflammation after intravitreal injection of adeno-associated virus (AAV) by slit-lamp or indirect ophthalmoscope. Here we evaluate the longitudinal profile of immediate inflammatory responses after AAV2 injection in rat retina and vitreous body by optical coherence tomography (OCT). Adult Fischer F344 rats were intravitreally injected once with saline, AAV2 or zymosan. Retinal thickness and cell infiltration were recorded by OCT longitudinally for 2 months and verified by histological analysis. The transduction rate of single intravitreal AAV2 injection was 21.3 ± 4.9% of whole retina, and the transduction efficiency on RGCs was 91.5 ± 2.5% in the transduced area. Significant increase in cell infiltration was observed from Day 1-3 after AAV2 injection, compared to very few infiltrating cells observed in the saline-injected group. The infiltrating cells ceased at Day 5 after intravitreal injection and remained absent at 2 months. The thicknesses of total and inner retina were increased along Day 1-3 after AAV2 injection, but reverted to normal afterwards. The surviving RGCs in the AAV2-injected groups at Day 14 showed no significant difference compared to saline-injected group. In summary, this study revealed the immediate inflammatory responses and retinal edema after intravitreal AAV2 injection in normal rats, without influencing long-term retinal thickness and RGC survival. OCT can be implemented for the time-lapse in vivo evaluation of inflammatory response after AAV-mediated gene therapy through intravitreal injection.

Casein kinase-II inhibition promotes retinal ganglion cell survival and axonal regeneration.

Neuron survival is critical for the maintenance of central nervous system physiology upon diseases or injury. We previously demonstrated that the blockage of phosphatidylinositol 3-kinase/Akt and Janus kinase/STAT3 pathways promotes retinal ganglion cell (RGC) survival and axonal regeneration via macrophage activation; yet, the complexity of the inflammatory regulation for neural repair indicates the involvement of additional unresolved signaling pathways. Here we report the effects and underlying mechanism of casein kinase-II (CK2) inhibition on RGC survival and axonal regeneration in rats after optic nerve (ON) injury. Adult rats received intravitreal injection of CK2 inhibitors, TBB (4,5,6,7-Tetrabromo-2-azabenzimidazole) and DMAT (2-Dimethylamino-4,5,6,7-tetrabromo-1H-benzimidazole), after ON transection and peripheral nerve (PN) grafting. Intravitreal application of TBB and DAMT effectively suppressed the CK2 phosphorylation activity in the retina, and enhanced RGC survival and axonal regeneration in vivo. Meanwhile, the numbers of infiltrating macrophages were increased. Removal of macrophages by clodronate liposomes significantly abolished the CK2 inhibition-induced RGC survival and axonal regeneration. Clodronate liposomes also weakened the RGC protective effects by TBB and DMAT in vitro. In summary, this study revealed that inhibition of CK2 enhances RGC survival and axonal regeneration via macrophage activation in rats. CK2 could be a therapeutic target for RGC protection after ON injury.

Improved performance of thermal-optic switch using polymer/silica hybrid and air trench waveguide structures.

By exploiting the polymer/silica hybrid and the air trench waveguide structures, we demonstrate a new type of low-power consuming and high-speed thermal-optic (TO) switch. Such a design provides an effective means to shorten the switching time of the TO switches, as well as to reduce the power consumption at the same time. This TO switch operated with less than 150 µs of switching time via a polymer/silica hybrid waveguide structure. Meanwhile, the power consumption was reduced to be 3.4 mW by introducing the air trench structure.

650-nm 1 × 2 polymeric thermo-optic switch with low power consumption.

In this paper, a low-power 1 × 2 polymeric thermo-optic switch operating at the polymer optical fiber low-loss window of 650 nm was studied. The characteristic parameters of the switch were carefully designed and simulated. The fabrication was done by using standard semiconductor fabrication techniques such as spin-coating, photolithography, and dry etching. The device was fabricated based on poly(methyl methacrylate) (PMMA)-based materials with the Mach-Zehnder interferometer (MZI) structure. The device shows an extinction ratio of over 23.4 dB at 650 nm with a very low-power consumption of 5.3 mW. The measured switching rise time and fall time are 464.4 and 448.0 µs, respectively.

The partner's insecure attachment, depression and psychological well-being as predictors of diurnal cortisol patterns for breast cancer survivors and their spouses.

The purpose of this study was to explore whether stress from individual's and partner's depression, anxiety, sleep disturbances, insecure attachment and meaning in life were predictors of diurnal cortisol patterns in breast cancer survivors and their spouses. Thirty-four couple dyads participated in this eight-month follow-up study. The breast cancer survivors and their spouses completed the Medical Outcomes Study Sleep scale, the Beck Depression Inventory-II, the State Trait Anxiety Inventory, the Experiences in Close Relationships-Revised scale and the Meaning in Life Questionnaire, and they collected salivary cortisol at home at the time of awakening, 30 and 45 min after waking and at 1200 h, 1700 h and 2100 h. Diurnal cortisol slopes of survivors and spouses are positively correlated. But the factors associated with diurnal cortisol patterns are different between survivors and spouses. For survivors, neither survivor individuals' nor spouses' psychosocial factors were the predictors of survivors' diurnal cortisol patterns. For spouses, the survivors' higher anxious attachment style was the main predictor of spouses' flatter diurnal cortisol patterns. In conclusion, for spouses, psychophysiological stress responses are mainly influenced by breast cancer survivors' insecure attachment. Future couple supportive care interventions can address survivors' attachment styles in close relationships in order to improve neuroendocrine functions for both breast cancer survivors and their spouses.

PEITC reverse multi-drug resistance of human gastric cancer SGC7901/DDP cell line.

Gastric cancer is one of the leading causes of cancer death in the world and nearly all patients who respond initially to cisplatin later develop drug resistance, indicating multi-drug resistance is an essential aspect of the failure of treatment. Phenethyl isothiocyanate (PEITC) has been implicated in inhibiting metastasis of several types of human cancer. However, the effect and potential mechanism of PEITC reversed multi-drug resistance of human gastric cancer is not fully clear. We have identified the role of PEITC in multi-drug resistance reversal of human gastric cancer SGC7901/DDP cell line. PEITC inhibited cisplatin-resistant human SGC7901/DDP cell growth in a dose-dependent manner, causing increased apoptosis, ROS generation, glutathione depletion, accumulation of Rhodamine-123, decreased expression of P-glycoprotein and cell cycle arrest. mRNA and protein expression of the multi-drug resistance gene (MDR1), multi-drug resistance-associated protein (MRP1), excision repair cross-complementing gene 1 (ERCC1), survivin, and Mad2 was decreased, and phosphorylation of Akt and transcriptional activation of NF-κB were suppressed. PEITC may be useful as the therapeutic strategy for overcoming multi-drug resistance through suppressing the PI3K-Akt pathway in human gastric cancer.

The effects of psychotherapy on psychological well-being and diurnal cortisol patterns in breast cancer survivors.

BACKGROUND: Neuroendocrine dysregulation influenced by psychosocial stress is related to breast cancer recurrence. Very few studies examine the impacts of psychotherapy on diurnal cortisol patterns among breast cancer survivors. METHODS: Forty-eight breast cancer patients who completed active cancer treatment were randomly assigned to receive either 8 weekly body-mind-spirit (BMS) group therapy sessions or 1 educational (EDU) session. Self-report measures included the Beck Depression Inventory-II (BDI-II), and the Meaning in Life questionnaire (MLQ) including two subscales: MLQ-Presence and MLQ-Search. Salivary cortisol levels were collected by the subjects in their homes at the time of awakening, 30 and 45 min after awakening, and at 12.00, 17.00, and 21.00 h. Measurement time points include baseline, the 2nd month (completion of BMS therapy), the 5th month, and the 8th month. RESULTS: There were no significant differences in BDI-II scores (p>0.05) and MLQ-Presence scores (p >0.05) between BMS and EDU groups at baseline or across the three follow-ups. Nevertheless, greater MLQ-Search scores were found in the BMS group compared to the EDU group during the 5th month of follow-up (p <0.01). The higher level of cortisol at 21.00 h (p < 0.01) and a flatter diurnal cortisol pattern were more likely to occur in EDU than in BMS participants (p < 0.05) at the 8th month of follow-up. CONCLUSION: BMS group therapy likely contributed to enhancing an active search for meaning in life toward more opportunities for personal growth and to maintaining stable cortisol responses to everyday life stress for breast cancer survivors.

Peritoneal macrophages attenuate retinal ganglion cell survival and neurite outgrowth.

Inflammation is a critical pathophysiological process that modulates neuronal survival in the central nervous system after disease or injury. However, the effects and mechanisms of macrophage activation on neuronal survival remain unclear. In the present study, we co-cultured adult Fischer rat retinas with primary peritoneal macrophages or zymosan-treated peritoneal macrophages for 7 days. Immunofluorescence analysis revealed that peritoneal macrophages reduced retinal ganglion cell survival and neurite outgrowth in the retinal explant compared with the control group. The addition of zymosan to peritoneal macrophages attenuated the survival and neurite outgrowth of retinal ganglion cells. Conditioned media from peritoneal macrophages also reduced retinal ganglion cell survival and neurite outgrowth. This result suggests that secretions from peritoneal macrophages mediate the inhibitory effects of these macrophages. In addition, increased inflammation- and oxidation-related gene expression may be related to the enhanced retinal ganglion cell degeneration caused by zymosan activation. In summary, this study revealed that primary rat peritoneal macrophages attenuated retinal ganglion cell survival and neurite outgrowth, and that macrophage activation further aggravated retinal ganglion cell degeneration. This study was approved by the Animal Ethics Committee of the Joint Shantou International Eye Center of Shantou University and the Chinese University of Hong Kong, Shantou, Guangdong Province, China, on March 11, 2014 (approval no. EC20140311(2)-P01).

CXCL5/CXCR2 modulates inflammation-mediated neural repair after optic nerve injury.

BACKGROUND: Previous studies reported that mild inflammation promotes retinal ganglion cell (RGC) survival and axonal regeneration after optic nerve (ON) injury with involvement of infiltrating macrophages and neutrophils. Here we aimed to evaluate the involvement and regulation of the main inflammatory chemokine pathway CXCL5/CXCR2 in the inflammation-mediated RGC survival and axonal regeneration in mice after ON injury. METHODS: The expressions and cellular locations of CXCL5 and CXCR2 were confirmed in mouse retina. Treatment effects of recombinant CXCL5 and CXCR2 antagonist SB225002 were studied in the explant culture and the ON injury model with or without lens injury. The number of RGCs, regenerating axons, and inflammatory cells were determined, and the activation of Akt andSTAT3 signaling pathways were evaluated. RESULTS: Cxcr2 and Cxcl5 expressions were increased after ON and lens injury. Addition of recombinant CXCL5 promoted RGC survival and neurite outgrowth in retinal explant culture with increase in the number of activated microglia, which was inhibited by SB225002 or clodronate liposomes. Recombinant CXCL5 also alleviated RGC death and promoted axonal regeneration in mice after ON injury, and promoted the lens injury-induced RGC protection with increase in the number of activated CD68+ cells. SB225002 inhibited lens injury-induced cell infiltration and activation, and attenuated the promotion effect on RGC survival and axonal regeneration through reduction of lens injury-induced Akt activation. CONCLUSIONS: CXCL5 promotes RGC survival and axonal regeneration after ON injury and further enhances RGC protection induced by lens injury with CD68+ cell activation, which is attenuated by CXCR2 antagonist. CXCL5/CXCR2 could be a potential therapeutic target for RGC survival promotion after ON injury.

Automatic detection of 39 fundus diseases and conditions in retinal photographs using deep neural networks.

Retinal fundus diseases can lead to irreversible visual impairment without timely diagnoses and appropriate treatments. Single disease-based deep learning algorithms had been developed for the detection of diabetic retinopathy, age-related macular degeneration, and glaucoma. Here, we developed a deep learning platform (DLP) capable of detecting multiple common referable fundus diseases and conditions (39 classes) by using 249,620 fundus images marked with 275,543 labels from heterogenous sources. Our DLP achieved a frequency-weighted average F1 score of 0.923, sensitivity of 0.978, specificity of 0.996 and area under the receiver operating characteristic curve (AUC) of 0.9984 for multi-label classification in the primary test dataset and reached the average level of retina specialists. External multihospital test, public data test and tele-reading application also showed high efficiency for multiple retinal diseases and conditions detection. These results indicate that our DLP can be applied for retinal fundus disease triage, especially in remote areas around the world.

Expression and characterization of thymine-DNA glycosylase from Aeropyrum pernix.

The recombinant thymine-DNA glycosylase (TDG) from Aeropyrum pernix (A. pernix) was expressed in Escherichia coli. The enzymatic activity of recombinant A. pernix TDG (ApeTDG) was characterized using oligonucleotides containing a thymine/uracil base as substrate. ApeTDG had distinct glycosylase activity on T/G mismatch. The optimal temperature and pH for thymine removal were 65-70 degrees C and pH 7.0-8.5, respectively. High concentration of NaCl inhibited the thymine removal. Divalent ions had different influence on the thymine removal by ApeTDG. Ca(2+) and Mg(2+) had no inhibition on the enzymic activity, but Ni(2+), Co(2+), Cu(2+), Mn(2+), and Zn(2+) completely inhibited the excision reaction. As derived from a hyperthermophilic archaea, ApeTDG protein was heat-resistant at 75 degrees C. ApeTDG also had a relatively weak DNA glycosylase activity on uracil base, with the following order: U/C>U/G approximately U/T>U/U approximately U/I approximately U/AP approximately U/->U/A. Additional mismatch located at 3' of T/G had less inhibition on the thymine removal than that located at 5' of T/G, and two additional mismatches located at each side of T/G completely inhibited the excision of thymine. Together, these data suggest that ApeTDG is a TDG protein with weak UDG activity.

Di-µ-chlorido-bis-[chlorido(N,N'-dibenzyl-propane-1,2-diamine-κN,N')copper(II)].

In the title complex, [Cu(2)Cl(4)(C(17)H(22)N(2))(2)], the Cu(II) cation is coordinated by a N,N'-dibenzyl-propane-1,2-diamine ligand and two Cl(-) anions, and a Cl(-) anion from an adjacent mol-ecule further bridges to the Cu(II) cation in the apical position, with a longer Cu-Cl distance of 2.9858 (18) Å, forming a centrosymmetric dimeric complex in which each Cu(II) cation is in a distorted square-pyramidal geometry. Intra-molecular N-H⋯Cl hydrogen bonding is observed in the dimeric complex.

Diaqua-bis(N,N'-dibenzyl-ethane-1,2-diamine-κN,N')nickel(II) dichloride N,N-dimethyl-formamide solvate.

The asymmetric unit of the title complex, [Ni(C(16)H(20)N(2))(2)(H(2)O)(2)]Cl(2)·C(3)H(7)NO, consists of two Ni(II) atoms, each lying on an inversion center, two Cl anions, two N,N'-dibenzyl-ethane-1,2-diamine ligands, two coordinated water mol-ecules and one N,N-dimethyl-formamide solvent mol-ecule. Each Ni(II) atom is six-coordinated in a distorted octa-hedral coordination geometry, with the equatorial plane formed by four N atoms and the axial positions occupied by two water mol-ecules. The complex mol-ecules are linked into a chain along [001] by N-H⋯Cl, N-H⋯O and O-H⋯Cl hydrogen bonds. The C atoms and H atoms of the solvent mol-ecule are disordered over two sites in a ratio of 0.52 (2):0.48 (2).

Bis{6,6'-dimeth-oxy-2,2'-[ethane-1,2-diyl-bis(imino-methyl-ene)]diphenolato(1.5-)-κO,N,N',O'}praeseodymium(III).

The title compound, [Pr(C(18)H(22.5)N(2)O(4))(2)], is isotypic with its Er and Tb analogues. All interatomic distances, angles and the hydrogen bond geometry are very similar for the three structures..

Bis{6,6'-dimeth-oxy-2,2'-[ethane-1,2-diyl-bis(imino-methyl-ene)]diphenolato(1.5-)-κO,N,N',O'}terbium(III).

The title compound, [Tb(C(18)H(22.5)N(2)O(4))(2)], is isotypic with its Pr and Tb analogues. All interatomic distances, angles and the hydrogen bond geometry are very similar for the three structures.

Type II collagen gene variants and inherited osteonecrosis of the femoral head.

BACKGROUND: Avascular necrosis of the femoral head (ANFH) causes disability that often requires surgical intervention. Most cases of ANFH are sporadic, but we identified three families in which there was autosomal dominant inheritance of the disease and mapped the chromosomal position of the gene to 12q13. METHODS: We carried out haplotype analysis in the families, selected candidate genes from the critical interval for ANFH on 12q13, and sequenced the promoter and exonic regions of the type II collagen gene (COL2A1) from persons with inherited and sporadic forms of ANFH. RESULTS: We identified a G-->A transition in exon 50 of COL2A1 in affected members of a four-generation family with ANFH. This transition predicts the replacement of glycine with serine at codon 1170 in a GXY repeat of type II collagen. Another pedigree was shown to harbor the same transition, but the mutant allele occurred on a different haplotype background. In a third family, a G-->A transition in exon 33 of the gene, causing a glycine-to-serine change at codon 717, was detected. No mutation was found in the COL2A1 coding region in sporadic cases of ANFH. CONCLUSIONS: All the patients with familial ANFH whom we studied carried COL2A1 mutations. In families with ANFH, haplotype and sequence analysis of the COL2A1 gene can be used to identify carriers of the mutant allele before the onset of clinical symptoms, allowing the initiation of measures that may delay progression of the disease.

Dysregulation of GIMAP genes in non-small cell lung cancer.

The GIMAP (GTPase of the immunity-associated protein) gene family includes seven functional members residing on human chromosome 7. GIMAP genes encode GTP-binding proteins that share a unique primary structure and whose function is largely unknown. However, gene ablation studies reveal that Gimap4 plays an important role in regulating the apoptosis of T cells. In a pilot microarray analysis on six cases of non-small cell lung cancer (NSCLC), we discovered that the expression of GIMAP family members, but not the neighboring non-GIMAP genes, was uniformly lower in the tumor tissues, compared to that in the adjacent nontumor tissues. This finding was subsequently confirmed by quantitative PCR assays in a total of twenty NSCLCs, and we found that GIMAP6 and GIMAP8 showed striking reduction of gene expression in the tumors. In contrast, GIMAP8 mRNA level was abnormally elevated in the adjacent nontumor tissues as compared to that in the control lung tissues. Such reciprocal expression of GIMAPs suggests that this unique gene family might contribute to the pathogenesis of and immune reactions to NSCLC.

A study on the efficacy of body-mind-spirit group therapy for patients with breast cancer.

AIMS AND OBJECTIVES: This study aims to understand the effects of culturally enriched body-mind-spirit group therapy on anxiety, depression and holistic well-being among women with breast cancer and to examine patients' views on what aspects of group therapy worked to enhance their health. DESIGN: The study was designed using multiple methods, which consisted of a randomised controlled trial and a focus group interview. METHODS: A total of 16 subjects in the control group received the standard care of a physician's treatment at the outpatient department. In addition to standard care, 12 subjects in the experimental group received 10 sessions of weekly body-mind-spirit group therapy for 180 minutes each. This therapy integrates concepts and practices of traditional Chinese medicine and Western medicine (e.g. positive psychology and forgiveness therapy). The subjects in the experimental group were invited to participate in a focus group interview regarding their perceptions of the change mechanisms that occurred in group therapy. RESULTS: The results of analysis of covariance indicated that after a two-month trial, there was a similarity between the experimental and control groups in reducing the scores of Beck depression inventory and increasing the scores of body-mind-spirit well-being. However, subjects in the experimental group had a better reduction of the scores of state anxiety inventory than subjects in the control group. The qualitative analysis yielded eight domains: (i) imparting of information, (ii) interpersonal learning, (iii) catharsis, (iv) universality, (v) group cohesiveness, (vi) altruism, (vii) instillation of hope and (viii) existential factors. These domains illustrate how the therapeutic effects of group therapy worked to reduce patients' anxiety. CONCLUSION: The culturally sensitive body-mind-spirit group therapy reduced anxiety among outpatients with breast cancer. RELEVANCE TO CLINICAL PRACTICE: The involvement of mental health nurses in providing group therapy for cancer patients could enhance the quality of care in psycho-ontological nursing.