Genetic Characteristics of Incidental Pheochromocytoma and Paraganglioma.

CONTEXT: Pheochromocytomas and paragangliomas (PPGLs) are being increasingly discovered by imaging performed for unrelated conditions. The genetic landscape of incidental PPGLs remains to be elucidated. OBJECTIVE: We aimed to describe the genetic characteristics of PPGLs discovered incidentally in a large PPGL cohort. METHODS: This retrospective cross-sectional study included 697 patients with pathology confirmed PPGLs, including 283 incidentalomas and 414 nonincidentalomas, at 2 tertiary care centers in China in 2009-2019. Tumor DNA samples were sequenced by next-generation sequencing. Identified genetic mutations were confirmed by Sanger sequencing and tested in 277 available matched blood DNA samples. RESULTS: There was a lower proportion of patients with mutations identified (53% vs 63.3%; P = 0.0067) in incidental than nonincidental PPGLs. In incidental PPGLs, HRAS (11.7%), FGFR1 (11%), and RET (9.2%) were the top 3 mutated genes, whereas HRAS (17.9%), VHL (9.2%), and NF-1 (8.7%) exhibited the highest rate of mutations in nonincidental PPGLs. In incidental pheochromocytomas, the most frequently mutated genes were RET (10.9%), HRAS (10.4%), and VHL (8.6%), while in incidental paragangliomas, FGFR1 (32.8%), HRAS (16.4%), and EPAS1 (9.8%) topped the list. The frequency of NF-1 mutations was significantly lower in incidental than nonincidental pheochromocytomas (4.1% vs 11%; P = 0.0042), while FGFR1 mutations were far more common in incidental than nonincidental paragangliomas (32.8% vs 15.3%; P = 0.0076). CONCLUSION: More than half of patients with incidental PPGLs had mutations in common susceptibility genes. The search for susceptibility genes should take both the mode of discovery (incidental vs nonincidental) and tumor location (pheochromocytoma vs paraganglioma) into consideration.

A Joint Lateral Motion-Stereo Constraint.

Purpose: We developed a stereo task that is based on a motion direction discrimination to examine the role that depth can play in disambiguating motion direction. Methods: In this study, we quantified normal adults' static and dynamic (i.e., laterally moving) stereoscopic performance using a psychophysical task, where we dichoptically presented randomly arranged, limited lifetime Gabor elements at two depth planes (one plane was at the fixation plane and the other at an uncrossed disparity relative to the fixation plane). Each plane contained half of the elements. For the dynamic condition, all elements were vertically oriented and moved to the left in one plane and to the right in another plane; for the static condition, the elements were horizontally oriented in one plane and vertically oriented in another plane. Results: For the range of motion speed that we measured (from 0.17°/s to 5.33°/s), we observed clear speed tuning of the stereo sensitivity (P = 3.0 × 10-5). The shape of this tuning did not significantly change with different spatial frequencies. We also found a significant difference in stereo sensitivity between stereopsis with static and laterally moving stimuli (speed = 0.67°/s; P = 0.004). Such difference was not evident when we matched the task between the static and moving stimuli. Conclusions: We report that lateral motion modulates human global depth perception. This motion/stereo constraint is related to motion velocity not stimulus temporal frequency. We speculate that the processing of motion-based stereopsis of the kind reported here occurs in dorsal extrastriate cortex.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) based assay for the simultaneous quantification of 18-hydroxycorticosterone, 18-hydroxycortisol and 18-oxocortisol in human plasma.

18-hydroxycorticosterone (18-OHB), 18-hydroxycortisol (18-OHF) and 18-oxocortisol (18-OXOF) are important biomarkers for the diagnosis of subtypes of primary aldosteronism. The detection of these three analytes by liquid chromatography-tandem mass spectrometry (LC-MS/MS) is free from structurally similar compounds. The aim of this study was to develop and validate a new LC-MS/MS assay for the simultaneous quantification of 18-OHB, 18-OHF and 18-OXOF in plasma and to establish a reference intervals for apparently healthy population. Plasma samples were prepared by solid phase extraction and separated in an ultra-high performance reversed phase column. MS detection was achieved using a triple quadrupole mass spectrometer in both positive and negative ionization modes. The developed assay was then validated against standard guidelines. We collected 691 plasma samples from apparently healthy individuals (M:398, F:293) to establish the reference intervals. The analytes were separated and quantified within 5 min. The newly developed method demonstrated linearity for the detected steroid concentration in range of 5 to 3000 pg/ml for 18-OXOF (r2 = 0.999) and 20 to 3000 pg/ml for 18-OHB (r2 = 0.997) and 18-OHF (r2 = 0.997). The lower limit of quantification (LLOQ) was 2.5 pg/ml, 20 pg/ml and 20 pg/m for 18-OXOF, 18-OHB and 18-OHF respectively. Specificity, precision, accuracy and stability were tested, and met the requirements of the guidelines. 18-OHB was higher in females than in males, but 18-OHF were higher in males than females. The reference intervals of 18-OHB, 18-OHF and 18-OXOF for both genders together were 90.5-1040.6 pg/ml, 224.4-1685.2 pg/ml, 4.0-70.5 pg/ml, respectively. Age was also an important factor influencing the levels of these three hormones. We have developed a sensitive and reliable method for the simultaneous quantification of 18-OHB, 18-OHF, and 18-OXOF. Our work provides a reference interval for the clinical application of these three steroid hormones.

A liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based assay for simultaneous quantification of aldosterone, renin activity, and angiotensin II in human plasma.

Accurate quantification of plasma aldosterone (ALD) and renin activity (PRA)is critical for the diagnosis of primary aldosteronism (PA). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) is considered the "gold standard" method for the determination of ALDand PRA. The aim of this study is to develop a new LC-MRM/MS assay for quantifying plasma ALD, PRA, and angiotensin II (Ang II) simultaneously and validate its effectiveness. To be more specific, plasmasamples were prepared by solid-phase extraction and separated in an ultra-performance reversed-phase column. MS detection was performed via a triple quadrupole mass spectrometer containing both positive and negative ion monitoring modes. The developed assay was then validated according to the standard guidelines and the influence of sample incubation on ALD and Ang II concentration was evaluated. In addition, the variation of endogenous Ang I was explored. The proposed LC-MRM/MS method was compared another LC-MS/MS method, which detects ALD, Ang I, and Ang II separately. Analyteswere separated and quantified within 5 min. The assay wasvalidated to be linear up to 5000 pg/ml for ALD and Ang II and 33.3 ng/ml/h for PRA.The lower limit of quantification (LLOQ) was 15 pg/ml, 15 pg/ml, and 0.1 ng/ml/hfor ALD, Ang II, and PRArespectively. Specificity, precision, accuracy, and stability were tested to meet the requirements of the guidelines. Significant changes were not found in ALD and Ang II concentrations over the 3 h-incubation. In addition, it was demonstratedthat the resultof PRA was not stronglyinfluenced by the endogenous Ang I. Comparison with another LC-MS/MS method was performed using the same apparatusand the proposed method was proved to be in good coincidence with the correlation coefficients rangingfrom 0.955to0.996. A sensitive and reliable method for simultaneousquantification of ALD, PRA, and Ang II has been developed and this study will significantly promote laboratory workflow efficiency and throughput.

Binocular visual deficits at mid to high spatial frequency in treated amblyopes.

Amblyopia (lazy eye) is a neurodevelopmental disorder of vision with no ocular pathology. The loss of vision in the amblyopic eye is assumed to be the main deficit in amblyopia, which has resulted in visual acuity (VA) being the primary outcome measure for treatment. Here we used a binocular orientation combination task to quantitatively assess the binocular status by measuring the binocular balance. We set out to determine whether amblyopes who reach the acuity-based end point have a residual binocular imbalance. Our results suggest that even amblyopes who have regained normal acuity have residual binocular deficits over a wide range of spatial frequencies. A further control study suggests that these binocular deficits could not be explained by any residual contrast sensitivity deficits of the amblyopic eye. Consequently, amblyopia is not the primary problem and VA is not the appropriate end point measure.

The Effects of 0.01% Atropine on Adult Myopes' Contrast Sensitivity.

PURPOSE: Atropine at a low concentration is considered a safe and effective treatment to mitigate myopia progression. However, the potential unwanted side effects of administering atropine at a low dose on visual functions other than best corrected visual acuity has not been investigated. In this study, we investigate the short-term (12,16, and 20 h) and long-term (1, 2, and 4 weeks) effects of 0.01% atropine (i.e., 0.1 mg/ml) on contrast sensitivity (CS) in patients with myopia. METHODS: Thirty adults (23.33 ± 2.93 years old) with myopia between -1.00 and -6.00 diopters (D), astigmatism of -1.50 D or less, and anisometropia of 1.00 D or less, participated in this prospective, masked, placebo-controlled, randomized study. The participants were randomly assigned to receive 0.01% atropine or polyvinyl alcohol eye drops once nightly to both eyes for four weeks. CS was measured binocularly at baseline and 12, 16, 20 h, 1, 2, and 4 weeks after the first use of the eye drops. RESULTS: There was no statistically significant differences of CS found between atropine and placebo-controlled groups in both short-term and long-term. There was no statistically significant interaction effect found between the time and group. CONCLUSION: We demonstrated no significant deleterious effect of 0.01% atropine on adult myopes' CS.

Smart Lipid-Based Nanosystems for Therapeutic Immune Induction against Cancers: Perspectives and Outlooks.

Cancer immunotherapy, a promising and widely applied mode of oncotherapy, makes use of immune stimulants and modulators to overcome the immune dysregulation present in cancer, and leverage the host's immune capacity to eliminate tumors. Although some success has been seen in this field, toxicity and weak immune induction remain challenges. Liposomal nanosystems, previously used as targeting agents, are increasingly functioning as immunotherapeutic vehicles, with potential for delivery of contents, immune induction, and synergistic drug packaging. These systems are tailorable, multifunctional, and smart. Liposomes may deliver various immune reagents including cytokines, specific T-cell receptors, antibody fragments, and immune checkpoint inhibitors, and also present a promising platform upon which personalized medicine approaches can be built, especially with preclinical and clinical potentials of liposomes often being frustrated by inter- and intrapatient variation. In this review, we show the potential of liposomes in cancer immunotherapy, as well as the methods for synthesis and in vivo progression thereof. Both preclinical and clinical studies are included to comprehensively illuminate prospects and challenges for future research and application.

Short-Term Deprivation Does Not Influence Monocular or Dichoptic Temporal Synchrony at Low Temporal Frequency.

Studies on binocular combination and rivalry show that short-term deprivation strengthens the contribution of the deprived eye in binocular vision. However, whether short-term monocular deprivation affects temporal processing per se is not clear. To address this issue, we conducted a study to investigate the effect of monocular deprivation on dichoptic temporal synchrony. We tested ten adults with normal vision and patched their dominant eye with an opaque patch for 2.5 h. A temporal synchrony paradigm was used to measure if temporal synchrony thresholds change as a result of monocular pattern deprivation. In this paradigm, we displayed two pairs of Gaussian blobs flickering at 1 Hz with either the same or different phased- temporal modulation. In Experiment 1, we obtained the thresholds for detecting temporal asynchrony under dichoptic viewing configurations. We compared the thresholds for temporal synchrony between before and after monocular deprivation and found no significant changes of the interocular synchrony. In Experiment 2, we measured the monocular thresholds for detecting temporal asynchrony. We also found no significant changes of the monocular synchrony of either the patched eye or the unpatched eye. Our findings suggest that short-term monocular deprivation induced-plasticity does not influence monocular or dichoptic temporal synchrony at low temporal frequency.

Aurora-A Induces Chemoresistance Through Activation of the AKT/mTOR Pathway in Endometrial Cancer.

Endometrial cancer (EC) is the most common gynecological tumor all over the world, and advanced/metastatic EC remains a malignancy with poor survival outcome due to highly resistant to conventional chemotherapeutic treatment. Here, we report that Aurora-A, a serine-threonine kinase, plays a vital role in chemoresistance of EC. Aurora-A is overexpressed in EC tissues, compared with normal endometrium and Aurora-A expression is associated with decreased overall survival. Overexpression of Aurora-A in EC cell lines (Ishikawa and HEC-1B cells) promotes cell proliferation and induced paclitaxel- and cisplatin-resistance. Furthermore, Aurora-A activating AKT-mTOR pathway further induces chemoresistance in vitro, consistent with a positive correlation between Aurora-A and phosphorylated AKT/4E-BP1 expression in EC tissues. In summary, our study provides the strong evidence that Aurora-A controls the sensitivity of EC cell lines to chemotherapy via AKT/mTOR pathway, indicating that pharmacologic intervention of Aurora-A and AKT/mTOR in combination with chemotherapy may be considered for the targeted therapy against EC with overexpression of Aurora-A.

Upregulation of PCP4 in human aldosterone-producing adenomas fosters human adrenocortical tumor cell growth via AKT and AMPK pathway.

Primary aldosteronism (PA) is characterized by aldosterone hypersecretion and adrenal hyperplasia and ranks as one of the most common causes of secondary hypertension. However, the molecular mechanism involved in adrenal hyperplasia and tumorigenesis is largely unknown. Dysregulation of Purkinji cell protein 4 (PCP4) is involved in the development and progression of neoplasia and aldosterone secretion, but little is known about the effect of PCP4 on human adrenocortical tumorigenesis. We investigated the expression pattern of PCP4 in different adrenal tissues and studied whether PCP4 is involved in cell growth in human adrenal cell lines. The mRNA levels of PCP4 were measured by real-time PCR in tissues from aldosterone-producing adenomas (APAs), idiopathic hyperaldosteronism (IHA) tissues, and normal adrenal (NA) tissues. In vitro siRNA knockdown of PCP4 in NCI-H295R and SW13 cell lines was used to determine the effect of PCP4 on cellular growth. Our results show that the mRNA level of PCP4 is upregulated in APAs and IHA compared with that in NA. The PCP4 mRNA expression level was positively correlated with tumor size in APAs. Knockdown of PCP4 decreased cell proliferation. Flow cytometry analysis showed that PCP4 knockdown fosters apoptosis. Finally, PCP4 knockdown inhibited phosphorylation of AKT308 and AMPKThr172. Our data suggest that PCP4 may represent a key player in the development and pathophysiology of PA via targeting the AKT and AMPK signaling pathways and thus may be a promising therapeutic target for PA.

Endocrinal description of two Chinese Kennedy's disease pedigrees.

Kennedy's disease (KD), also known as X-linked spinal and bulbar muscular atrophy (SBMA), is caused by the expansion of cytosine-adenine-guanine (CAG) repeats in the first exon of the androgen receptor (AR) gene. KD is a late-onset neural-endocrinal disease that is characterized by the degeneration of motor neurons in the brainstem and spinal cord. In addition, partial androgen insensitivity is an important manifestation of KD. Here, we report two Chinese KD pedigrees that reveal the clinical and genetic manifestations and fully elaborate the endocrinal characteristics of KD patients. The proband in pedigree 1 was referred to an endocrinologist for gynaecomastia and sexual dysfunction. A gene analysis of this patient revealed that there were 53 CAG repeats in the AR gene. A family survey identified an additional two KD patients in pedigree 1. The proband in pedigree 2 was diagnosed by a neurologist and did not have gynaecomastia or sexual dysfunction. A family survey identified an additional subclinical patient, and both patients exhibited partial androgen insensitivity at a hormonal level. We therefore suggest that a family survey and hormone tests should be routinely performed in KD patients and that physicians should increase their understanding of the different symptoms of KD to achieve correct diagnoses in affected patients.