Ovarian granulosa cell tumors frequently express EGFR (Her-1), Her-3, and Her-4: An immunohistochemical study

OBJECTIVE: Up to 50% of patients with ovarian granulosa cell tumors (GCTs) will develop recurrences; some of these recurrences can be seen as late as 30 years following the initial surgical treatment. Combined chemotherapy and radiotherapy are currently used for patients with advanced or recurrent disease. The aim of this study was to investigate the possible eligibility of patients with GCTs for anti-Her therapy. METHODS: The immunohistochemical expression of EGFR (Her-1), Her-2, Her-3, and Her-4 was analyzed in a group of ovarian GCTs encompassing 38 adult type and 2 juvenile type. RESULTS: Thirty-one cases (77.5%) were positive for at least one of the receptors EGFR (Her-1), Her-3, and Her-4. Twenty-six out of 40 (65%) GCTs showed positive reaction for EGFR (Her-1). Eight tumors (20%) were exclusively positive for EGFR (Her-1). None of 40 cases showed a positive reaction for Her-2. Positive reactions for Her-3 and Her-4 were observed in 18 (45%) and 23 (57.5%) tumors. Only one case (2.5%) was exclusively positive for Her-4. Four tumors (10%) showed positivity for Her-3 and Her-4 but were negative for EGFR (HER-1). While one of the two JGCTs was negative for all members of the Her-family, one showed reactivity for EGFR (Her-1), Her-3, and Her-4. CONCLUSION: In this study, most of the ovarian GCTs express at least one of the receptors EGFR (Her-1), Her-3, and Her-4. These findings provide some evidence to further explore the potential use of agents targeting these receptors (particularly EGFR) in the treatment of ovarian GCTs.     

Model examination of chemoprevention with retinoids in squamous cell carcinomas of the head and neck region and suitable biomarkers for chemoprevention

The prognosis for patients with head and neck tumors (HNSCC) is poor, due among other things to the high-risk factor for locoregional recurrence and/or second primary tumors. Extensive studies on chemoprevention of oral pre-cancers to stop carcinogenesis and to prevent recurrence and/or second primary tumors have failed to reach the desired effects. The toxicity of retinoids (RA) for example limits their dosage. Biomarkers are used to evaluate the duration of therapy. In this study, cell culture models are used to demonstrate immunocytochemical expression of RA receptors (RAR, RXR), Ki-67 and p53 before and after all-trans retinoic acid (ATRA) treatment. Telomerase activity in PCR is used to assess the effectiveness of ATRA. Along with an RA-sensitive HNSCC cell line UM-SCC-35 we employed cell lines UM-SCC-14C and HaCaT. Our immunocytochemical examination produced no proof of a statistically significant change in expression of RARα, RARβ or RXRγ receptors after ATRA treatment, either in the cells of the sensitive UM-SCC-35 line or in HaCaT cells. The RARβ and RXRγ receptors showed increased expression after brief cell treatment of UM-SCC-14C. The reduced telomerase activity after prolonged treatment of the UM-SCC-35-cells with ATRA (as well as the reduced p53 expression) proved to be a biomarker for evaluating the success of therapy. Although XTT and MTT tests demonstrated that cell proliferation in UM-SCC-35 cells was inhibited after brief and extended RA influence, the immunocytological Ki-67 scores failed to confirm the inhibition. No reduction of p53 expression, of telomerase activity or of cell proliferation in the XTT and MTT test was detected in the RA-insensitive cell line UM-SCC-14C or in HaCaT cells. We also demonstrated the parameters used in examining the models in sections of carcinoma tissue and in control tissues from the head and neck region, so they can be examined in clinical chemopreventive studies on biopsy tissue.     

Synthesis and DNA transfection properties of new head group modified malonic acid diamides

The goals of the current study were to develop and characterize a nanoemulsion of ezetimibe, evaluate its stability, lipid lowering and pharmacokinetic profile. Solubility of the drug was estimated in various oils and surfactants. Existence of nanoemulsion region was confirmed by plotting phase diagrams. Various thermodynamic stability and dispersibility tests were performed on the formulations chosen from phase diagram. Percentage transmittance, refractive index, viscosity, droplet size and zeta potential of the optimized formulations were determined. Dialysis bag method was employed to study the release rate. The formulation selected for bioavailability estimation contained Capryol 90 (10%, v/v), Crempophor EL (11.25%, v/v), Transcutol^® P (33.75%, v/v), and double distilled water (45%, v/v). The release rate from the nanoemulsion was highly significant (p < 0.001) in contrast to the drug suspension. The level of total cholesterol in the group receiving nanoemulsion CF1 was found to be highly significant (p < 0.001) in comparison to the group receiving drug suspension. Bioavailability studies in rats revealed superior absorption of ezetimibe from nanoemulsion as compared to the marketed formulation and drug suspension. The shelf life of the nanoemulsion was estimated to be 18.53 months. The present study corroborated nanoemulsion to be a promising choice to improve the bioavailability of ezetimibe.     

Fatal Cryptococcus gattii genotype AFLP6/VGII infection in a HIV‐negative patient: case report and a literature review

Cryptococcus gattii, a species belonging to the Cryptococcus complex which occurs endemically in tropical and subtropical regions, has been reported as a causative agent of cryptococcosis in healthy individuals. We report a case of meningitis in HIV‐negative patient from Cuiaba, MT, in the Midwestern region of Brazil. Cryptococcus gattii AFLP6/VGII was isolated from cerebrospinal fluid and molecular typing was performed by URA5‐RFLP. The in vitro susceptibility profile was determined using the standard method according to the document M27A3, CLSI 2008. C. gattii AFLP6/VGII was shown to be susceptible to the antifungals tested. Treatment with 0.8 mg/kg of amphotericin B was initiated; however, the patient died 2 days after the onset of therapy.     

Further studies of the norditerpene (+)-harringtonolide isolated from Cephalotaxus harringtonia var. drupacea: absolute configuration, cytotoxic and antifungal activities

Harringtonolide (= hainanolide) is a complex polycyclic fused norditerpene isolated from CEPHALOTAXUS HARRINGTONIA var. DRUPACEA. In spite of its appealing biological properties - we measured an IC (50) of 43 nM on KB cells and a significant antifungal activity - its absolute configuration has not yet been firmly established. This was done herein using X-ray anomalous scattering after bromination of the tropone ring, unambiguously giving the stereochemistry 5 R,6 R,7 S,13 S,14 S,15 R,16 R. Detailed IN VITRO biological measurements are provided.     

A New Environmentally Friendly Approach to Lignin‐Based Cyclic Carbonates

Herein, a novel synthesis of a fully renewable lignin‐based building block equipped with cyclic carbonate functionalities is presented. In an efficient two‐step procedure, organosolv lignin from ethanol–water pulping of beech wood is oxyalkylated with glycerol carbonate to insert adjacent hydroxyl groups. The oxyalkylated lignin is then reacted via transesterification with dimethyl carbonate or ethylene carbonate in dimethyl sulfoxide under alkaline conditions producing cyclic carbonate functionalized lignin. This transesterification is studied as a function of time, catalyst type, and catalyst amount using NMR spectroscopy as well as size exclusion chromatography. Transesterification reactions with K₂CO₃ as catalyst (0.4 eq.) afford lignin‐based cyclic carbonate with almost complete conversion (96%) within 6 h, as analyzed by ³¹P and ¹³C NMR spectroscopy. This green building block represents a reactive prepolymer for the synthesis of nonisocyanate polyurethanes.     

Dysregulated immunity in PID patients with low GARP expression on Tregs due to mutations in LRRC32

Immune dysregulation diseases are characterized by heterogeneous clinical manifestations and may have severe disease courses. The identification of the genetic causes of these diseases therefore has critical clinical implications. We performed whole-exome sequencing of patients with immune dysregulation disorders and identified two patients with previously undescribed mutations in LRRC32, which encodes glycoprotein A repetitions predominant (GARP). These patients were characterized by markedly reduced numbers and frequencies of regulatory T cells (Tregs). Tregs with mutated LRRC32 exhibited strongly diminished cell-surface GARP expression and reduced suppressor function. In a model of conditional Garp deficiency in mice, we confirmed increased susceptibility to inflammatory diseases once GARP expression on Tregs was decreased. Garp deficiency led to an unstable Treg phenotype due to diminished Foxp3 protein acetylation and stability. Our study reinforces the understanding of the immunological mechanisms of immune dysregulation and expands the knowledge on the immunological function of GARP as an important regulator of Treg stability.     

Apoptotic cells enhance sphingosine‐1‐phosphate receptor 1 dependent macrophage migration

The lipid sphingosine‐1‐phosphate (S1P) is a chemokine for a variety of immune cells including lymphocytes and monocytes. Migration toward S1P is determined by the S1P receptor expression profile, with S1PR1/3 (where S1PR is S1P receptor) stimulating and S1PR2 attenuating migration. However, the impact and physiological significance of S1P‐induced migration of macrophages is largely unclear. We observed that alternative activation of human macrophages, by IL‐4 or apoptotic cells (ACs), enhanced S1PR1 expression. Moreover, ACs provoked macrophage migration toward S1P in an S1PR1‐dependent manner as confirmed by pharmacological receptor inhibition and S1PR1‐deficient murine macrophages. In a mouse model of resolving peritoneal inflammation, F4/80‐driven deletion of S1PR1 reduced postinflammatory macrophage emigration from inflammatory sites. S1PR1 expression on macrophages might, therefore, be relevant for restoring tissue homeostasis during the resolution of inflammation.     

The intravenous perfluorocarbon emulsion Oxycyte does not increase hyperbaric oxygen-related seizures in a non-sedated swine model

Purpose

Standard treatment for decompression sickness (DCS) is recompression therapy with hyperbaric oxygen (HBO). Non-recompressive therapies are needed to address mass casualty scenarios such as a disabled submarine rescue or DCS therapy in remote environments. Intravenously delivered perfluorocarbon (PFC) emulsions improve blood oxygen content and decrease mortality in several animal models of DCS. However, the enhanced oxygen delivery of PFC emulsions may increase CNS oxygen toxicity (seizures) risk when used in conjunction with HBO. We studied seizure latency and duration in swine randomized to receive PFC or normal saline with 6 ATA of oxygen.

Methods

Yorkshire swine (n = 31) were fitted with EEG electrodes and randomized to receive 5 ml/kg of the PFC Oxycyte (Oxygen Biotherapeutics Inc., Morrisville, NC) or saline intravenously 1 h before HBO. Unsedated animals were fitted with a snout mask for inhaled gas delivery, positioned inside the hyperbaric chamber, and compressed to 165 ft of sea water (6 ATA). After 2.5 min at 6 ATA, breathing gas was switched to 100 % O₂ until signs of seizure were observed and EEG activity was evident. At seizure onset gas was switched back to air for 3 min, then the chamber was decompressed. After 24 h, the dive profile/oxygen exposure was repeated to ensure no secondary effects of PFC drug redistribution or emulsion metabolism.

Results/conclusion

Intravenous PFC emulsion did not decrease seizure latency or increase duration on initial HBO exposure or after 24 h. This finding demonstrates the safety of PFC use in conjunction with recompression therapy to treat DCS.