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El cambio de color arlequín es un trastorno benigno, idiopático y autolimitado que se caracteriza por una apariencia de la piel dividida en dos zonas de color distinto. Su etiología es desconocida, pero se cree que está causada por una inmadurez hipotalámica del tono vascular periférico. La infección por COVID-19 en neonatos es infrecuente y raramente sintomática, con sólo unos pocos casos descritos en la literatura. De forma aislada, ambas afecciones tienen una baja incidencia. Este es el primer caso descrito en la literatura mundial de cambio de coloración arlequín en un recién nacido que dio positivo a COVID-19. Aun no existe ninguna publicación que relacione el cambio de color arlequín con COVID-19.

One novel brominated nocardiopsistin D (

Background: Harlequin color change is a benign, idiopathic, self-limiting disorder characterized by an appearance of skin divided into two distinctly colored areas. Its etiology is unknown but thought to be caused by immaturity of hypothalamic regulation of peripheral vascular tone. COVID-19 infection in neonates is infrequent and rarely symptomatic, with only a few cases described in the literature. In isolation, both conditions have low incidence. This is the first case reported in the world literature of harlequin color change in a newborn who tested positive for COVID-19. There isn’t a single publication that links harlequin color change to COVID-19, yet. The Case: Newborn of a 34-year old mother. Who at 3 hours of life presented progressive respiratory distress with whimpering and cyanosis. At 20 hours presented a sudden color change in the right hemibody with unilateral hyperemia and contralateral pallor, simultaneous with a demarcated line separating the left and right sides of the body, involving the trunk, extremities and face, this remained for 10 minutes and then began to fade on its own until it disappeared completely. There were no changes in vital parameters associated with this phenomenon. Only one episode was reported. The transient character and the strict unilateral distribution of the manifestations did not confirm any etiology other than Harlequin phenomenon. Subsequently the patient evolved to respiratory deterioration, multiorgan failure (SOFA: 21), died at 22 days of life due to respiratory arrest. Conclusion: This is the first ever documented case in worldwide literature of harlequin color change in a neonate positive to COVID-19. Given the low incidence of both entities in isolation and the fact that they were discovered in the same patient, we suggest a strong possibility of SARS-CoV-2 to be the source of the harlequin phenomena in this case and should be considered as an indicator of severe COVID in newborns, but further studies that expand the selection of patients with this condition are needed. There is no literature linking the harlequin phenomenon with COVID-19.

Introducción: en 1952, Neligan y Strange describieron por primera vez un fenómeno raro, que se produce en el periodo neonatal y se caracteriza por un cambio repentino del color de la piel, con una línea recta que divide la mitad del cuerpo como si hubiera sido "dibujada con un lápiz y una regla" que discurre por la línea media, presentando la mitad del cuerpo eritematoso y la otra pálida. Objetivo: presentar literatura relacionada con el fenómeno del cambio de color arlequín y sus diferentes presentaciones clínicas. Métodos: se llevó a cabo una revisión no sistemática de la literatura disponible utilizando palabras claves como “Harlequin Color Change”, “Harlequin Newborn”, “Harlequin Color Change NOT Syndrome NOT ichthyosis” además de sinónimos, que se combinaron con los conectores “AND” y “OR”, tanto en inglés como en español, en las bases de datos PubMed, ScienceDirect y Google Academics. Resultados: el cambio de color arlequín se caracteriza por un aspecto de la piel dividida en dos zonas de color distinto. Se desconoce su etiología, pero se cree que está causado por inmadurez de la regulación hipotalámica del tono vascular periférico. Afecta aproximadamente 10% de los neonatos sanos. Conclusiones: el cambio de color arlequín es una condición benigna, idiopática y autolimitada que no necesita tratamiento. El médico debe reconocer este fenómeno, que presenta una variedad de patrones de cambio de color. Es esencial informar a los padres de esta afección cutánea benigna y transitoria para evitar pruebas y tratamientos innecesarios. No debe confundirse con la ictiosis arlequín.

Cancer cells grown in spheroid conditions interact with each other and the extracellular matrix, providing a better representation of the in vivo environment than two-dimensional cultures and are a more clinically relevant model. A discrete screening of genetically diverse marine samples in the spheroid assay led to the identification of a novel activity for the known compound furospinulosin 1. This compound shows activity against MDA-MB-231 triple negative breast cancer cells grown as spheroids and treated for 24 or 48 h. No cytotoxicity was seen in traditional two-dimensional adherent cultures treated for a longer time (72 h). A reverse phase protein array (RPPA) confirmed the limited activity of the compound in cells grown traditionally and revealed changes in protein expression when cells are grown as spheroids that are associated with better clinical prognosis. Analysis of the RPPA data through the Broad institute’s connectivity map suggested the hypothesis that furospinulosin 1 functions as an MEK inhibitor. Analysis of the RPPA data through STRING supports the apoptosis observed. The selectivity exhibited by furospinulosin 1 for triple negative breast cancer cells only when grown as spheroids makes it an interesting compound with strong therapeutic potential that merits further study.

Membrane trafficking is essential for all cells, and visualizing it is particularly useful for studying neuronal functions. Here we report the synthesis, characterization, and application of several membrane- and pH-sensitive probes suitable for live-cell fluorescence imaging. These probes are based on a 1,8-naphthalimide fluorophore scaffold. They exhibit a solvatochromic effect, and one of them, ND6, shows a substantial fluorescence difference between pH 6 and 7. The solvatochromic effect and pH-sensitivity of those probes are explained using quantum chemical calculations, and molecular dynamics simulation confirms their integration and interaction with membrane lipids. For live-cell fluorescence imaging, we tested those probes in a cancer cell line (MCF7), cancer spheroids (MDA-MB-468), and cultured hippocampal neurons. Confocal imaging showed an excellent signal-to-noise ratio from 400:1 to about 1300:1 for cell membrane labeling. We applied ND6 during stimulation to label nerve terminals via dye uptake during evoked synaptic vesicle turnover. By ND6 imaging, we revealed cholesterol's multifaced role in replenishing synaptic vesicle pools. Our results demonstrate these fluorescent probes' great potential in studying membrane dynamic and synaptic functions in neurons and other secretory cells and tissues.

Survivin is a 16.5 KDa protein whose functions include promoting cellular mitosis, angiogenesis, and senescence as well as inhibiting apoptosis. Higher survivin expression is found in cancer tissues than normal tissues, and this expression correlates with disease progression and aggressiveness. Survivin has been validated as a clinical target for cancer. Small molecules are important antagonists of survivin levels in cancer cells. A structurally diverse library of genetically encoded small molecules (natural products) derived from marine plants, invertebrates, and microbes was screened for their ability to reduce expression levels of survivin in the DLD-1 colon adenocarcinoma and the A549 nonsmall cell lung carcinoma cell lines. This led to the identification of this novel activity for the known compounds eryloside E, ilicicolin H, tanzawaic acid A, and p-hydroxyphenopyrrozin. Both eryloside E and ilicicolin H showed the ability to reduce survivin expression in the low micromolar range against both cell lines.

Cryptosporidium sp. are apicomplexan parasites that cause significant morbidity and possible mortality in humans and valuable livestock. There are no drugs on the market that are effective in the population most severely affected by this parasite. This study is the first high-throughput screen for potent anti-Cryptosporidium natural products sourced from a unique marine compound library. The Harbor Branch Oceanographic Institute at Florida Atlantic University has a collection of diverse marine organisms some of which have been subjected to medium pressure liquid chromatography to create an enriched fraction library. Numerous active compounds have been discovered from this library, but it has not been tested against Cryptosporidium parvum. A high-throughput in vitro growth inhibition assay was used to test 3764 fractions in the library, leading to the identification of 23 fractions that potently inhibited the growth of Cryptosporidium parvum. Bioassay guided fractionation of active fractions from a deep-sea sponge, Leiodermatium sp., resulted in the purification of leiodolide A, the major active compound in the organism. Leiodolide A displayed specific anti-Cryptosporidium activity at a half maximal effective concentration of 103.5 nM with selectivity indexes (SI) of 45.1, 11.9, 19.6 and 14.3 for human ileocecal colorectal adenocarcinoma cells (HCT-8), human hepatocellular carcinoma cells (Hep G2), human neuroblastoma cells (SH-SY5Y) and green monkey kidney cells (Vero), respectively. The unique structure of leiodolide A provides a valuable drug scaffold on which to develop new anti-Cryptosporidium compounds and supports the importance of screening natural product libraries for new chemical scaffolds.

Pancreatic cancer, the fourth leading cause of cancer death in the United States, has a negative prognosis because metastasis occurs before symptoms manifest. Although combination therapies are showing improvements in treatment, the survival rate for pancreatic cancer five years post diagnosis is only 8%, stressing the need for new treatments. The receptor for advanced glycation end products (RAGE) has recently emerged as a chemotherapeutic target in KRAS driven pancreatic cancers both for treatment and in chemoprevention. RAGE appears to be an important regulator of inflammatory, stress and survival pathways that lead to carcinogenesis, resistance to chemotherapy, enhanced proliferation and the high metastatic potential of pancreatic cancer. RAGE expression has been demonstrated in pancreatic cancer tumors but not in adjacent epithelial tissues. Its presence is associated with increased proliferation and metastasis. In an effort to identify novel inhibitors of RAGE among our collection of marine-derived secondary metabolites, a cell-based screening assay utilizing flow cytometry was developed. This effort led to the identification of scalarin as the active compound in a marine sponge identified as Euryspongia cf. rosea. Scalarin is a sesterterpene natural product isolated previously from a different marine sponge. Scalarin reduces the levels of RAGE and inhibits autophagy in the PANC-1 and MIA PaCa-2 pancreatic cancer cell lines. Its IC50 for cytotoxicity ranges between 20 and 30 μM in the AsPC-1, PANC-1, MIA PaCa-2 and BxPC-3 pancreatic cancer cell lines. Inhibition of autophagy limits tumor growth and tumorigenesis in pancreatic cancer, making scalarin an interesting compound that may merit further study.

Pancreatic cancer presents one of the most negative prognosis of all cancers as it has usually metastasized by the time a patient is diagnosed. The American Cancer Society estimates that 93% of patients will die within 5 years of diagnosis, highlighting the need for new drugs to treat this disease. Interleukin 8 (IL-8) mediates the angiogenesis of tumors arising from Ras mutations, which are present in about 90% of pancreatic adenocarcinomas. Overexpression of IL-8 in pancreatic tumors is believed to promote tumor angiogenesis and to activate survival signaling pathways. A 96-well cell-based enzyme-linked immunosorbent assay was set up to screen the Harbor Branch Oceanographic Institute library of marine natural products to identify those with the ability to inhibit IL-8 production by BxPC-3 pancreatic cancer cells. Over 1000 fractions were screened, resulting in the identification of 10 known marine natural products with this ability. These compounds fall into four classes of compounds including the pyrroloiminoquinone alkaloids secobatzelline A and isobatzelline C; mycalamide A and B, onnamide A, discalamide A, and theopederin K from the mycalamide class of polyketides; the lipopeptide microcolin A; and the cyclic depsipeptides didemnin B and nordidemnin B. In addition, didemnin B, nordidemnin B, and theopederin K induce potent cytotoxicity against four pancreatic cancer cell lines tested. Many of these compounds have been previously reported to inhibit protein synthesis and the decrease in IL-8 production may be nonspecific. Nevertheless, this is a new activity for these compounds and inhibition of IL-8 secretion by pancreatic cancer cells can now be added to the previously reported antiangiogenic activities of the didemnins.

Our understanding of cell death used to consist in necrosis, an unregulated form, and apoptosis, regulated cell death. That understanding expanded to acknowledge that apoptosis happens through the intrinsic or extrinsic pathways. Actually, many other regulated cell death processes exist, including necroptosis, a regulated form of necrosis, and autophagy-dependent cell death. We also understand that apoptosis occurs beyond the intrinsic and extrinsic pathways with caspase independent forms of apoptosis existing. Our knowledge of the signaling continues to grow, and with that, so does our ability to target different parts of the pathways with small molecules. Marine natural products co-evolve with their targets, and these unique molecules have complex structures with exquisite biological activities and specificities. This article offers a review of our current understanding of the signaling pathways regulating cell death, and highlights marine natural products that can affect these signaling pathways.

Harmful algal blooms that can produce toxins are common in the Indian River Lagoon (IRL), which covers ~250 km of Florida's east coast. The current study assessed the dynamics of microcystins and saxitoxin in six segments of the IRL: Banana River Lagoon (BRL), Mosquito Lagoon (ML), Northern IRL (NIRL), Central IRL (CIRL), Southern IRL (SIRL), and the St. Lucie Estuary (SLE). Surface water samples (n = 40) collected during the 2018 wet and 2019 dry season were analyzed to determine associations between toxins and temperature, salinity, pH, oxygen saturation, concentrations of dissolved nutrients and chlorophyll-a, presence of biosynthetic genes for toxins, relative abundance of planktonic species, and composition of the microbial community. The potential toxicity of samples was assessed using multiple mammalian cell lines. Enzyme-Linked Immunosorbent Assays were used to determine concentrations of microcystins and saxitoxin. Overall, the microcystins concentration ranged between 0.01-85.70 µg/L, and saxitoxin concentrations ranged between 0.01-2.43 µg/L across the IRL. Microcystins concentrations were 65% below the limit of quantification (0.05 µg/L), and saxitoxin concentrations were 85% below the limit of detection (0.02 µg/L). Microcystins concentrations were higher in the SLE, while saxitoxin was elevated in the NIRL and BRL. Cytotoxicity related to the presence of microcystins was seen in the SLE during the wet season. No significant patterns between cytotoxicity and saxitoxin were identified. Dissolved nutrients were identified as the most highly related parameters, explaining 53% of microcystin and 47% of saxitoxin variability. Multivariate models suggested cyanobacteria, flagellates, ciliates, and diatoms as the subset of microorganisms whose abundances were maximally correlated with saxitoxin and microcystins concentrations. Lastly, biosynthetic genes for microcystins were detected in the SLE and for saxitoxin in the BRL and NIRL. These results highlight the synergistic roles environmental and biological parameters play in influencing the dynamics of toxin production by harmful algae in the IRL.

Pancreatic cancer, the fourth leading cause of cancer death in the United States, has a negative prognosis because metastasis occurs before symptoms manifest. Leiodermatolide, a polyketide macrolide with antimitotic activity isolated from a deep water sponge of the genus Leiodermatium, exhibits potent and selective cytotoxicity toward the pancreatic cancer cell lines AsPC-1, PANC-1, BxPC-3, and MIA PaCa-2, and potent cytotoxicity against skin, breast and colon cancer cell lines. Induction of apoptosis by leiodermatolide was confirmed in the AsPC-1, BxPC-3 and MIA PaCa-2 cells. Leiodermatolide induces cell cycle arrest but has no effects on in vitro polymerization or depolymerization of tubulin alone, while it enhances polymerization of tubulin containing microtubule associated proteins (MAPs). Observations through confocal microscopy show that leiodermatolide, at low concentrations, causes minimal effects on polymerization or depolymerization of the microtubule network in interphase cells, but disruption of spindle formation in mitotic cells. At higher concentrations, depolymerization of the microtubule network is observed. Visualization of the growing microtubule in HeLa cells expressing GFP-tagged plus end binding protein EB-1 showed that leiodermatolide stopped the polymerization of tubulin. These results suggest that leiodermatolide may affect tubulin dynamics without directly interacting with tubulin and hint at a unique mechanism of action. In a mouse model of metastatic pancreatic cancer, leiodermatolide exhibited significant tumor reduction when compared to gemcitabine and controls. The antitumor activities of leiodermatolide, as well as the proven utility of antimitotic compounds against cancer, make leiodermatolide an interesting compound with potential chemotherapeutic effects that may merit further research.

Lobomycosis (lacaziosis) is a chronic, granulomatous, fungal infection of the skin and subcutaneous tissues of humans and dolphins. To date, the causative agent, the yeast-like organism Lacazia loboi, has not been grown in the laboratory, and there have been no recent reports describing attempts to culture the organism. As a result, studies on the efficacy of therapeutics and potential environmental reservoirs have not been conducted. Therefore, the objective of the current study was to utilize both classical and novel microbiological methods in order to stimulate growth of Lacazia cells collected from dolphin lesions. This included the experimental inoculation of novel media, cell culture, and the use of artificial skin matrices. Although unsuccessful, the methods and results of this study provide important insight into new approaches that could be utilized in future investigations of this elusive organism.

Two new analogues of the potent antitumor compound leiodermatolide, which we call leiodermatolides B and C, have been isolated from specimens of a deep-water sponge of the genus Leiodermatium collected off Florida. The compounds were purified using standard chromatographic methods, and the structures defined through interpretation of the HRMS and 1D and 2D NMR data. Leiodermatolide B (2) lacks the C-21 hydroxy group found in leiodermatolide and has equal potency as the parent compound, providing a simpler analogue for possible clinical development. It inhibits the proliferation of the AsPC-1 human pancreatic adenocarcinoma cell line with an IC50 of 43 nM. Leiodermatolide C (3) has a modified macrolide ring and is over 85-fold less potent with an IC50 of 3.7 μM against the same cell line. These compounds add to the knowledge of the pharmacophore of this class of potent antitumor agents.

A deep-water sponge of the genus Spongosorites has yielded a bis-indole alkaloid which we have named dragmacidin G. Dragmacidin G was first reported by us in the patent literature and has recently been reported by Hitora et al. from a sponge of the genus Lipastrotheya. Dragmacidin G is the first in this series of compounds to have a pyrazine ring linking the two indole rings. It also has a rare N-(2-mercaptoethyl)-guanidine side chain. Dragmacidin G shows a broad spectrum of biological activity including inhibition of methicillin-resistant Staphylococcus aureus, Mycobacterium tuberculosis, Plasmodium falciparum, and a panel of pancreatic cancer cell lines.

Este trabajo es resultado de la inves­tigación que implicó la búsqueda, procesamiento, contrastación, análi­sis y evaluación de documentación archi­vada, junto con la que se recabó en fuentes secundarias, que dan cuenta del desarrollo del currículo en la formación de maestras para la atención de párvulos en la primera Escuela Normal de Educadoras en Chihua­hua en 1929. Se retoma la Historia Social como metodología para desarrollar esta investigación, cuyo propósito general se centró en valorar los planes de estudio a través de las siguientes preguntas: ¿Cómo se visualizaba la formación docente? ¿Cuá­ les eran las ideas en torno a la formación de educadoras en el estado de Chihuahua en 1929? ¿Cuáles fueron los planes de estudio implementados?

La inserción de la mujer en el mercado laboral y el reclamo de la igualdad de derechos y obligaciones de la mujer, subyugada al poder varonil por centurias, trajo en el mundo la necesidad de conformar una cultura igualitaria. Proceso nada sencillo, que dentro de su complejidad históricamente se ha venido construyendo de manera asimétrica entre las diferentes regiones del país. Aquí se trata de ofrecer algunos elementos de ese andamiaje político-académico y cultural desde la perspectiva estadística.

Pancreatic cancer, the fourth leading cause of cancer death in the United States, has a negative prognosis because metastasis occurs before symptoms manifest. Leiodermatolide, a polyketide macrolide with antimitotic activity isolated from a deep water sponge of the genus Leiodermatium, exhibits potent and selective cytotoxicity towards the pancreatic cancer cell lines AsPC-1, PANC-1, BxPC-3, and MIA PaCa-2, and potent cytotoxicity against skin, breast and colon cancer cell lines. Induction of apoptosis by leiodermatolide was confirmed in the AsPC-1, BxPC-3 and MIA PaCa-2 cells. Leiodermatolide induces cell cycle arrest but has no effects on in vitro polymerization or depolymerization of tubulin alone, while it enhances polymerization of tubulin containing microtubule associated proteins (MAPs). Observations through confocal microscopy show that leiodermatolide, at low concentrations, causes minimal effects on polymerization or depolymerization of the microtubule network in interphase cells, but disruption of spindle formation in mitotic cells. At higher concentrations, depolymerization of the microtubule network is observed. Visualization of the growing microtubule in HeLa cells expressing GFP-tagged plus end binding protein EB-1 showed that leiodermatolide stopped the polymerization of tubulin. These results suggest that leiodermatolide may affect tubulin dynamics without directly interacting with tubulin and hint at a unique mechanism of action. In a mouse model of metastatic pancreatic cancer, leiodermatolide exhibited significant tumor reduction when compared to gemcitabine and controls. The anti-tumor activities of leiodermatolide, as well as the proven utility of anti-mitotic compounds against cancer, make leiodermatolide an interesting compound with potential chemotherapeutic effects that may merit further research.

A través del tiempo la mujer como profesional de la educación ha venido adquiriendo mayor presencia en la historia de la educación preescolar en Chihuahua. Por ello en el presente artículo se analiza la manera en que algunas de las mujeres, fueron asumiendo el papel de maestras para la atención de la primera infancia, selladas con mayor relevancia en la historiografía decimonónica al generalizarse su presencia como maestras reconocidas socialmente, hasta las primeras décadas del siglo XX, periodo en el que se crea la primera Escuela Normal de Educadoras en Chihuahua de la que egresan un gran número de señoritas que se desempeñan con calidad en su labor profesional y establecen las bases sólidas de la educación preescolar en el estado. El escrito se aborda a través de las siguientes categorías: panorama general del papel desempeñado por las mujeres chihuahuenses, desempeño de las educadoras chihuahuenses en el mercado laboral y el futuro de la carrera de educadoras en Chihuahua.

The age-related macular degeneration (ARMD) is a bilateral maculopathy that affect central vision and has a social, scientific and economic repercussion due to the visual discapacity that causes.With the objective of characterizing the behaviour of the AMD, it was done a study of 49 patients that went to the retina consultation of the oftalmological service from the General Hospital Camilo Cienfuegos of Sancti Spíritus the period of january 1 st , 2006 to december 31 st 2008. Different variables were used like: age, sex, skin color, risk factors, general deseases, clinical forms and treatment. It prevailed the male sex (53,0%), the age group of 80 years plus (57,1 %), the white race (93,8%), the arterial hypertension as a general desease (38,7%), and the dry clynical form (89.7%). It was found the advance age and the white race as main risk factors and prevailed the dry clinic form of the desease.

Leiodermatolide is a structurally unique macrolide, isolated from the deep-water marine sponge Leiodermatium sp., which exhibits potent antiproliferative activity against a range of human cancer cell lines (IC50

We found two errors in our previous published paper [1]. Figure 4A has a mistake in the units in the labels, where it shows mM instead of micromolar (µM). A correctly labeled Figure 4A ensues. In Figures 2 and 4, the size bar scale is micrometers (µm). We apologize for the inconvenience caused to our readers. [...]

Purpose: Pancreatic cancer is the fourth leading cause of cancer death in the United States, and new drugs to treat the disease are needed. Pancreatic cancer cells are highly metastatic and exhibit resistance to apoptosis. Small molecules that can restore sensitivity to apoptosis or reduce metastasis would have therapeutic potential against this disease. Manzamine A is an alkaloid isolated from marine sponges that was suspected to have inhibitory activity against the mitogen activated kinase kinase (MEK). Because of this, the effects of Manzamine A were studied in pancreatic cancer cells. Methods: AsPC-1 cells were treated for 48 h in the presence of various concentrations of Manzamine A and their phenotype, cytotoxicity, cell invasion and susceptibility to apoptosis were observed. Results: Manzamine A decreased single cell formation, abrogated cell migration and restored the susceptibility of the cells to TRAIL-induced apoptosis in AsPC-1 cells. Its mechanism of action remains unknown, as manzamine A does not inhibit MEK. Conclusions: Manzamine A appears to have a formerly unrecognized activity in blocking tumor cell invasion as well as in restoring cancer cell susceptibility to apoptosis in vitro and therefore has the potential to be used as an adjuvant to existing cancer therapies.

Microphthalmia-associated transcription factor (Mitf) is essential for melanocyte development and function and regulates anti-apoptotic Bcl2 expression. We hypothesized that cellular deficiency of Mitf can influence melanocyte survival in response to ultraviolet (UV) radiation. Primary melanocyte cultures were prepared from neonatal wild-type mice and congenic animals heterozygous for Mitf mutations Mitf (mi-vga9/+) and Mitf(Mi-wh/+) and exposed to UV irradiation. Wild-type melanocytes were more resistant to UV-induced apoptosis than melanocytes partially deficient in Mitf activity, as determined by relative levels of intracellular melanin and relative activation of Mitf target genes Tyr, Tyrp1, Dct, and Cdk2. Comparative experiments with wild-type cells and congenic albino melanocytes demonstrated that these differences are not due to differences in melanin content, implicating Mitf as a primary determinant of UV-dependent melanocyte survival. Mitf activity correlated directly with resistance to UV-induced apoptosis in melanocytes. Mitf was important not only for regulating the expression of anti-apoptotic Bcl-2 following UV irradiation, but also the expression of the pro-apoptotic BH3-only Bad protein and activation of the extrinsic apoptotic pathway. Hence, Mitf is a multifaceted regulator of UV-induced apoptosis in melanocytes.

A new marine-derived macrolide designated as neopeltolide (1) has been isolated from a deep-water sponge of the family Neopeltidae. Its structure was elucidated on the basis of spectroscopic data interpretation. Neopeltolide (1) is a potent inhibitor of the in vitro proliferation of the A-549 human lung adenocarcinoma, the NCI-ADR-RES human ovarian sarcoma, and the P388 murine leukemia cell lines, with IC50's of 1.2, 5.1, and 0.56 nM, respectively. Neopeltolide (1) also inhibited the growth of the fungal pathogen Candida albicans with a minimum inhibitory concentration of 0.62 microg/mL.

Background/purpose: Ultraviolet (UV) radiation is animportant therapy for immune-mediated cutaneousdiseases. Activation of early apoptotic pathways mayplay a role in the clinical effectiveness. Different UVwavelengths have different efficacy for various dis-eases, but it remains unclear whether the ability toinduce apoptosis differs with respect to the wavelength,and whether they induce apoptosis through the samemechanism. The aim of this study is to analyze theeffects of different UV wavelengths that are usedclinically on normal human peripheral blood mono-nuclear cells (PBMCs).Methods: PBMCs were treated with UV-light sourcesbroadband UVB, narrowband UVB, broadband UVAand UVA1. Initiation of apoptosis was assessed byflow cytometry by staining–treated cells for activatedcaspases. Immunoblots were performed to measure forcleaved caspase-3, -8, -9, cytochrome c, Bcl 2-inter-acting domain and poly-(ADP ribose) polymerasecleavage.Results: We demonstrate that all the UV radiationsources induced caspase activation in a dose-andtime-dependent manner. Components of both the ex-trinsic and intrinsic pathways of apoptosis were acti-vated by all of the UV wavelengths tested, but differedin the level of energy needed for activation.Conclusion: The greater effectiveness of UVB oninitiation of apoptotic pathway suggests that apoptosismay play a role in the clinical efficacy of UVB-responsive inflammatory cutaneous diseases.Keywords: apoptosis; caspase; lymphocytes; peripheralblood mononuclear cells; ultraviolet light.

Se realizó un estudio prospectivo longitudinal en el Hospital Universitario “Camilo Cienfuegos “ de Sancti Spíritus en el periodo comprendido del 1ro de septiembre del 2003 al 31 de agosto del 2004 con el objetivo de conocer las complicaciones trans-operatorias y post-operatorias de la cirugía de catarata así como el impacto de esta nueva técnica a través del grado de satisfacción de la población con los servicios quirúrgicos recibidos. El universo de la investigación estuvo integrado por los 663 pacientes que fueron llevados al quirófano para realizarle la técnica quirúrgica de extracción extracapsular del cristalino con implante de Lente Intraocular. Fue la catarata senil la de mayor incidencia (90,79). La patología ocular previa mas asociada a la catarata en estos pacientes fue la Miopía (5,63%). Las complicaciones trans operatorias fueron las más frecuentes, predominando la ruptura de la cápsula posterior (9,95%) acompañada de Vítreo Grado I en el mayor número de casos (86,36%). Las complicaciones post-operatorias inmediatas y mediatas tuvieron una baja incidencia, no así las tardías que ocurrieron en el 7,99% de los casos, el 76,62% de los operados alcanzaron visión por encima de 20/60. El grado de satisfacción de los pacientes operados, con el servicio quirúrgico recibido fue bueno en el 99,8%.

Se realizó un estudio prospectivo longitudinal acerca del comportamiento clínico – epidemiológico de la baja visión en Sancti Spiritus, que abarcó un estudio de 5 años (2001- 2005).El universo estuvo integrado por 227 pacientes, de los cuales 186 eran adultos y 103 correspondieron al sexo masculino. Las patologías más frecuentes que causaron baja visión fueron la degeneración macular senil, miopía, retinopatía diabética, glaucoma, atrofia óptica y retinosis pigmentaria. Se logró rehabilitar al 82.8 % de los pacientes.

Se realizó un estudio descriptivo que incluyó a tres miembros de una familia, comenzando por una paciente, su hijo y un nieto, todos con una atrofia óptica dominante, con daños visuales variables en dependencia del progreso de la enfermedad, y por lo tanto con necesidad de ayudas ópticas o no. Se tomaron fotos del fondo de ojo de cada paciente, y se presentan como un caso de gran interés oftalmológico, seguido incluso por la consulta de genética en nuestra provincia.

Apoptosis plays a critical role in the development and progression of ultraviolet-induced skin cancers. In particular, Fas and Fas ligand (FasL) interactions are known to control the development of “sunburn cells” or apoptotic keratinocytes in the UV-exposed epidermis. In the absence of functional Fas/FasL signaling, UV-induced apoptosis is diminished and mutations rapidly accumulate. UV-induced suppression of host immunity, a process regulating skin cancer outgrowth, is also controlled through Fas/FasL interactions. Other death receptors, such as the receptor for tumor necrosis factor, may also contribute to UV-induced carcinogenesis and progression. Understanding the involvement of cell death in cancers caused by exposure to sunlight may provide novel approaches for prevention and therapy of these ever-increasing malignancies.

Pancreatic cancer, the 4th leading cause of cancer death in the US, is highly resistant to all current chemotherapies, and its growth is facilitated by chronic inflammation. An important mediator of inflammation is the nuclear factor kappa B (NFκB), a transcription factor that regulates over 500 genes including the regulation of anti-apoptotic proteins, cell cycle progression and cytokine production. NFκB is constitutively activated in pancreatic cancer cells contributing to their resistance to apoptosis and high metastatic potential. Although many small molecules that inhibit NFκB have been identified, none are currently used in the clinic, perhaps due to their lack of specificity. To identify novel inhibitors of NFκB, the HBOI library of enriched fractions from marine organisms was screened using a reporter cell line that produces luciferin under the transcriptional control of NFκB. Fractions from the sponge Amphibleptula were active in this screen and contained the antifungal cyclic peptide microsclerodermin A. Microsclerodermin A is shown here to inhibit NFκB transcriptional activity in a reporter cell line, to reduce levels of phosphorylated (active) NFκB in the AsPC-1 cell line, to have an IC50 for cytotoxicity in the low micromolar range against the AsPC-1, BxPC-3, MIA PaCa-2 and PANC-1 pancreatic cancer cell lines, and to induce significant apoptosis in the AsPC-1, BxPC-3 and the PANC-1 cell lines. Treatment of AsPC-1 cells with microsclerodermin A also resulted in an increase in IL-8 production without apparent induction of angiogenic factors and there is the possibility that inhibition of NFκB by microsclerodermin A is mediated by the glycogen synthase kinase 3β pathway.

A hybrid library of the marine natural products dictyostatin and discodermolide, incorporating the taxol or taxotere side chains, were synthesised; preliminary biological evaluation in the PANC-1 cancer cell line revealed significant antiproliferative activity, demonstrating that a macrolide scaffold is an effective surrogate for the baccatin core of taxol.

Manzamine A, a member of the manzamine alkaloids, was originally isolated from marine sponges of the genus Haliclona. It was recently shown to have activity against pancreatic cancer cells, but the precise mechanism of action remained unclear. To further our understanding of the mechanism of action of manzamine A, chemogenomic profiling in the yeast S. cerevisiae was performed, suggesting that manzamine A is an uncoupler of vacuolar ATPases. Fluorescence microscopy confirmed this effect on yeast vacuoles, where manzamine A produced a phenotype very similar to that of the established v-ATPase inhibitor bafilomycin A1. In pancreatic cancer cells, 10 µM manzamine A affected vacuolar ATPase activity and significantly increased the level of autophagosome marker LC3-II and p62/SQSTM1 as observed by western blot analysis. Treatment with manzamine A in combination with bafilomycin A1 (inhibitor of autophagosome-lysosome fusion) did not change the levels of LC3-II when compared to cells treated with bafilomycin A1 alone, suggesting that manzamine A is a potential inhibitor of autophagy by preventing autophagosome turnover. As autophagy is essential for pancreatic tumor growth, blocking this pathway with manzamine A suggests a promising strategy for the treatment of pancreatic cancer.

Pancreatic cancer, the fourth leading cause of cancer death in the US, is highly resistant to all current chemotherapies, and its growth is facilitated by chronic inflammation. The majority of pro-inflammatory cytokines initiate signaling cascades that converge at the activation of the Nuclear Factor Kappa B (NFκB), a signal transduction molecule that promotes cell survival, proliferation and angiogenesis. In an effort to identify novel inhibitors of NFκB, the HBOI library of pure compounds was screened using a reporter cell line that produces luciferin under the transcriptional control of NFκB. Seven compounds were identified through this screen, but in the case of five of them, their reported mechanism of action made them unlikely to be specific NFκB inhibitors. Spongiatriol, a marine furanoditerpenoid that was first isolated in the 1970s, is shown here to inhibit NFκB transcriptional activity in a reporter cell line, to reduce levels of phosphorylated (active) NFκB in the AsPC-1 cell line, to have an IC50 for cytotoxicity in the low micromolar range against the AsPC-1, BxPC-3, MiaPaCa-2 and Panc-1 pancreatic cancer cell lines, and to induce moderate but significant apoptosis in both the AsPC-1 and the Panc-1 cell lines.

Background and Aims: Vacuolar ATPases are proton transporters located in the membranes of intracellular compartments such as lysosomes, which play an important role in the autophagic pathway. Vacuolar ATPases play a crucial role in drug resistance and tumor metastasis, by changing the pH gradient between intracellular organelles and the cytoplasma or between extracellular and intracellular compartments. Thus, vacuolar ATPases may represent a novel target for anti-cancer strategies. Manzamine A, a marine sponge derived alkaloid, was recently shown to have activity against pancreatic cancer cells. To elucidate the mechanism of action of manzamine A, its effect on vacuolar ATPases and the autophagic pathway was studied. Methods: The HIP/HOP (haploinsufficency profiling/homozygous profiling) assay to chemogenomically profile manzamine A was conducted at Novartis. Results suggested that manzamine A is an uncoupler of vacuolar ATPases. To confirm this function, expression of vacuolar ATPases in pancreatic cancer cell lines was analyzed by immunocytochemistry in the presence or absence of manzamine A. To evaluate the effect of manzamine A on vacuolar ATPase proton pump activity/ lysosomal acidification, cells were stained with Lysosensor pH indicator followed by detection of acidic lysosomes by immunoflorescent staining and flow cytometry. To analyze the effect of manzamine A on autophagy, pancreatic cancer cells were treated with manzamine A in presence or absence of the autophagy inhibitor bafilomycin, followed by the detection of the autophagy markers, Light Chain 3 (LC 3-II) and Sequestosome 1 (p62/ SQSTM1) by Western blot analysis. Results: Treatment of four pancreatic cancer cell lines (AsPC-1, MIA-PaCa2, BxPc-3, Panc-1) with manzamine A had no effect on expression of vacuolar ATPases. However, it resulted in an increase of intra-lysosomal acidity, which suggests interference with proton pump activity of vacuolar ATPases. Manzamine A significantly increased the LC 3-II and p62/SQSTM1 levels in AsPC-1 cells. Treatment with manzamine A in combination with bafilomycin A1 did not change the levels of LC 3-II when compared to cells treated with bafilomycin A1 alone, indicating that manzamine A is a potential inhibitor of autophagy (inhibitor of autophagosome turnover). Conclusion: Results indicate that manzamine A interferes with proton pump activity of vacuolar ATPases in pancreatic cancer cells, induces an increase of intralysosomal acidity, and affects autophagosome turnover, making it a potential inhibitor of autophagy likely to act at the level of autophagosome-lysosome fusion. Citation Format: Georgios Kallifatidis, Dominic Hoepfner, Sven Schuierer, Nicole Hartmann, Esther Guzman, Amy Wright. The marine natural product manzamine A targets vacuolar ATPases and autophagy in pancreatic cancer cells. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Progress and Challenges; Jun 18-21, 2012; Lake Tahoe, NV. Philadelphia (PA): AACR; Cancer Res 2012;72(12 Suppl):Abstract nr A39.

Two new marine-derived sesquiterpene benzoquinones which we designate as neopetrosiquinones A (1) and B (2), have been isolated from a deep-water sponge of the family Petrosiidae. The structures were elucidated on the basis of their spectroscopic data. Compounds 1 and 2 inhibit the in vitro proliferation of the DLD-1 human colorectal adenocarcinoma cell line with IC50 values of 3.7 and 9.8 μM, respectively, and the PANC-1 human pancreatic carcinoma cell line with IC50 values of 6.1 and 13.8 μM, respectively. Neopetrosiquinone A (1) also inhibited the in vitro proliferation of the AsPC-1 human pancreatic carcinoma cell line with an IC50 value of 6.1 μM. The compounds are structurally related to alisiaquinone A, cyclozonarone, and xestoquinone.

The design, synthesis and biological evaluation of a series of hybrids and analogues of the microtubule-stabilising anticancer agents dictyostatin, discodermolide and taxol is described. A 22-membered macrolide scaffold was prepared by adapting earlier synthetic routes directed towards dictyostatin and discodermolide, taking advantage of the distinctive structural and stereochemical similarities between these two polyketide-derived marine natural products. Initial endeavours towards accessing novel discodermolide/dictyostatin hybrids led to the adoption of a late-stage diversification strategy and the construction of a small library of methyl ether derivatives, along with the first triple hybrids bearing the side chain of taxol or taxotere attached through an ester linkage. Following biological assays of the antiproliferative activity of these compounds in a series of human cancer cell lines, including the taxol-resistant NCI/ADR-Res cell line, the results allowed the proposal of various structure-activity relationships. This led to the identification of a potent macrocyclic discodermolide/dictyostatin hybrid 12 and its C9 methoxy derivative 38, accessible by an efficient total synthesis and having a similar biological profile to dictyostatin.

Lasonolide A, a novel polyketide-derived macrolide, was previously identified from an extract of the marine sponge Forcepia sp. in an assay for protein kinase C (PKC) inhibitors. Cytotoxicity testing and profiling of lasonolide A in the National Cancer Institute (NCI) 60 cell panel screen revealed that it was potent toward a broad range of cell lines and also suggested a unique mechanism of action. Contrary to expected results, we found lasonolide A to be a strong activator of PKC in Panc-1 pancreatic carcinoma cells. Downstream mitogen-activated protein kinases, ERK 1/2 and p38 were also rapidly phosphorylated in response to lasonolide A, as was Akt. Microscopy studies revealed that lasonolide A induced blebbing and contraction of the cells within minutes of exposure, and the eventual loss of adherence. However, membrane integrity was maintained and the effects were reversible if lasonolide A was washed from the cells after their loss of adherence. Pretreatment of cells with a myosin II inhibitor, blebbistatin, slowed the early onset, but did not prevent the morphological effects of lasonolide A. Cells stained for actin filaments showed some reduction in stress fiber structure after lasonolide A exposure; however, it did not affect the polymerization of purified actin in vitro. Bisindolemaleimide, a PKC inhibitor, and wortmannin, a phosphoinositide 3-kinase; inhibitor, did not reduce lasonolide A-induced contraction or blebbing or the activation of mitogen-activated protein kinases, although Akt phosphorylation was prevented by wortmannin pretreatment. Our results indicate that lasonolide A activates multiple signal transduction pathways and suggest that the origin is upstream of PKC.

A new adenine substituted bromotyrosine derived metabolite designated as aphrocallistin (1) has been isolated from the deep-water Hexactinellida sponge Aphrocallistes beatrix beatrix Gray, 1858 (Order Hexactinosida, Family Aphrocallistidae). Its structure was elucidated on the basis of spectral data and confirmed through a convergent, modular total synthetic route that is amenable towards future analog preparation. Aphrocallistin inhibits the growth of a panel of human tumor cell lines with IC50 values ranging from 7.5 to >100 μM and has been shown to induce G1 cell cycle arrest in the PANC-1 pancreatic carcinoma cell line. Aphrocallistin has been fully characterized in the NCI cancer cell line panel and has undergone in vitro ADME pharmacological profiling.

BackgroundMetalloproteinase inhibitors can protect mice against experimental autoimmune encephalomyelitis (EAE), an animal model for multiple sclerosis (MS). Matrix metalloproteinase-9 (MMP-9) has been implicated, but it is not clear if other MMPs are also involved, including matrilysin/MMP-7 – an enzyme capable of cleaving proteins that are essential for blood brain barrier integrity and immune suppression.ResultsHere we report that MMP-7-deficient (mmp7-/-) mice on the C57Bl/6 background are resistant to EAE induced by myelin oligodendrocyte glycoprotein (MOG). Brain sections from MOG-primedmmp7-/-mice did not show signs of immune cell infiltration of the CNS, but MOG-primed wild-type mice showed extensive vascular cuffing and mononuclear cell infiltration 15 days after vaccination. At the peak of EAE wild-type mice had MMP-7 immuno-reactive cells in vascular cuffs that also expressed the macrophage markers Iba-1 and Gr-1, as well as tomato lectin. MOG-specific proliferation of splenocytes, lymphocytes, CD4+and CD8+cells were reduced in cells isolated from MOG-primedmmp7-/-mice, compared with MOG-primed wild-type mice. However, the adoptive transfer of splenocytes and lymphocytes from MOG-primedmmp7-/-mice induced EAE in naïve wild-type recipients, but not naïvemmp7-/-recipients. Finally, we found that recombinant MMP-7 increased permeability between endothelial cells in anin vitroblood-brain barrier model.ConclusionOur findings suggest that MMP-7 may facilitate immune cell access or re-stimulation in perivascular areas, which are critical events in EAE and multiple sclerosis, and provide a new therapeutic target to treat this disorder.

Pancreatic cancer is the fourth leading cause of cancer death in the United States. The prognosis of the disease is very negative, because the cancer will be usually metastasized by the time a patient manifests symptoms. Although combination therapy shows some promise, new drugs to treat the disease are needed. Given our interest in finding new therapies for pancreatic cancer, we sought to determine whether the known cytotoxic activity of the batzellines extended to pancreatic cancer cell lines. The batzellines are pyrroloiminoquinones alkaloids obtained from the deep-water Caribbean sponge Batzella sp (family Esperiopsidae, order Poecilosclerida). We show here that batzellines exhibit selective cytotoxicity towards the pancreatic cancer cell lines AsPC-1, Panc-1, BxPC-3, and MIA PaCa2 compared with the normal African green monkey kidney epithelial cell line Vero. The batzellines cause cytotoxicity by inducing cell cycle arrest that is mediated by their ability to intercalate into DNA and/or inhibit topoisomerase II activity. The cytotoxic abilities of isobatzellines A and C against pancreatic cancer cell lines, their low toxicity against normal cells, and their reported ability to be synthesized makes them interesting compounds with potential chemotherapeutic effects that may merit further research.

By exploiting a Still–Gennari olefination of a common C11–C26 aldehyde with a C4–C10 or C1–C10 β-ketophosphonate, three modified C2–C6 region analogues of the 22-membered macrolide dictyostatin were synthesised and evaluated in vitro for growth inhibition against a range of human cancer cell lines, including the Taxol-resistant NCI/ADR-Res cell line. 6-Desmethyldictyostatin and 2,3-dihydrodictyostatin displayed potent (low nanomolar) antiproliferative activity, intermediate between dictyostatin and discodermolide, while 2,3,4,5-tetrahydrodictyostatin showed activity comparable to discodermolide. As with dictyostatin, these simplified analogues act through a mechanism of microtubule stabilisation, G2/M arrest and apoptosis.

By exploiting a Still-Gennari HWE coupling with a common C11-C26 aldehyde, a series of C2-C6 modified analogues of the microtubule-stabilising marine natural product dictyostatin were synthesised and evaluated in vitro for growth inhibition against a range of human cancer cell lines, including the (P-glycoprotein efflux-mediated) Taxol-resistant NCI/ADR cell line. Removal of the C6 methyl substituent in dictyostatin was found to be well tolerated and led to the retention of antiproliferative activity in the low nanomolar range (IC(50)=43 nM in the NCI/ADR cell line), while partial and full saturation of the (2Z,4E)-dienoate region led to a progressive reduction in biological potency. The lactone ring size was found to be critical, as C21 to C19 translactonisation to afford 20-membered isodictyostatin analogues led to a significant loss of cytotoxicity. In a series of incubatory experiments performed on the PANC-1 cell line, all three of the 22-membered macrolide analogues acted in an analogous fashion to dictyostatin, through a mechanism of microtubule stabilization, causing both an accumulation of cells at the G2/M phase and formation of characteristic dense intracellular microtubule bundles.

It is well established that females mount stronger immune responses than males, but only very little is understood about the underlying mechanisms. We have analyzed local cytokine differences among intact females, those that had been ovariectomized (OVX), those receiving estrogen replacement after OVX, and males, both before and after production of delayed-type hypersensitivity (DTH) responses. We report confirmation of a much larger DTH response in females versus males. However, OVX resulted in an even larger response, while estrogen replacement resulted in a smaller response when compared with intact females. In animals exposed for the first time to an antigen (without a DTH response), OVX increased interleukin-6 (IL-6) and estrogen replacement after OVX suppressed IL-6. Of the cytokines that differed between males and females exposed for the first time to an antigen, only IL-6 was higher in females versus males when exposure to antigen occurred for the second time (when the DTH response occurs). Analysis of cytokines with OVX and estrogen replacement after a second exposure to antigen showed that IL-6 did not significantly change. Levels of IL-4; Regulated upon Activation, Normal T-cell Expressed; and Secreted; and thrombopoietin, however, correlated with the DTH response, suggesting direct or indirect positive regulation by estrogen. These results suggest an important role for both IL-6 and IL-4 in determining the degree of DTH response, with IL-6 (which appears negatively regulated by estrogen) increasing and IL-4 (which appears positively regulated by estrogen) decreasing the response. The results further suggest that IL-6 may play a role in predisposing to a larger DTH response, while IL-4 levels seem more important during an active response.

Pancreatic cancer presents one of the most negative prognoses of all cancers, as it has usually metastasized by the time a patient is diagnosed. Therefore, almost as many deaths as new cases are expected each year. Although combination therapy shows some promise, the need for new drugs to treat the disease is pressing. Harbor Branch Oceanographic Institute (HBOI) has developed a unique library of marine samples and its research program seeks to tap into the biological diversity of the oceans to find potential anti-cancer drugs. Inflammation has been connected to pancreatic cancer development and progression. A recent study has shown that IL-8 mediates the angiogenesis of tumors arising from Ras-mutations. Ras is a proto-oncogene that is mutationally activated in 90% of pancreatic adenocarcinomas. Overexpression of IL-8 in pancreatic tumors is thought to promote tumor angiogenesis and to activate survival signaling pathways. Given our interest in finding new therapies for pancreatic cancer, we set up a 96-well cell-based assay to screen our library of marine natural products for those with the ability to inhibit IL-8 production by pancreatic cancer cells. A thousand fractions were screened resulting in the identification of marine natural products with the ability to inhibit IL-8 secretion by the BxPC-3 pancreatic cancer cell line. Studies are currently underway to determine if this inhibition leads to an inhibition in angiogenesis or affects cell proliferation. Citation Format: Esther A. Guzman, Tara Pitts, Amy E. Wright. Inhibition of IL-8 production by pancreatic cancer cells with marine natural products. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr A50.