Pharmacology of boldine: summary of the field and update on recent advances.

Over the past decade, boldine, a naturally occurring alkaloid found in several plant species including the Chilean Boldo tree, has garnered attention for its efficacy in rodent models of human disease. Some of the properties that have been attributed to boldine include antioxidant activities, neuroprotective and analgesic actions, hepatoprotective effects, anti-inflammatory actions, cardioprotective effects and anticancer potential. Compelling data now indicates that boldine blocks connexin (Cx) hemichannels (HCs) and that many if not all of its effects in rodent models of injury and disease are due to CxHC blockade. Here we provide an overview of boldine's pharmacological properties, including its efficacy in rodent models of common human injuries and diseases, and of its absorption, distribution, pharmacokinetics, and metabolism.

Pathophysiological role of connexin and pannexin hemichannels in neuromuscular disorders.

A growing body of research has provided evidence that de novo expression of connexin hemichannels and upregulation of pannexin hemichannels (Cx HCs and Panx HCs, respectively) in the cytoplasmic membrane of skeletal muscle (sarcolemma) are critical steps in the pathogenesis of muscle dysfunction of many genetic and acquired muscle diseases. This review provides an overview of the current understanding of the molecular mechanisms regulating the expression of Cx and Panx HCs in skeletal muscle, as well as their roles in both muscle physiology and pathologies. Additionally, it addresses existing gaps in knowledge and outlines future challenges in the field.

Unilateral Hypofunction of the Masseter Leads to Molecular and 3D Morphometric Signs of Atrophy in Ipsilateral Agonist Masticatory Muscles in Adult Mice.

Mice are commonly used to study mandibular dynamics due to their similarity in chewing cycle patterns with humans. Adult mice treated unilaterally with botulinum toxin type A (BoNTA) in the masseter exhibit atrophy of this muscle characterized by an increase in the gene expression of atrophy-related molecular markers, and a reduction in both muscle fiber diameter and muscle mass at 14d. However, the impact of this muscle imbalance on the non-treated masticatory muscles remains unexplored. Here, we hypothesize that the unilateral masseter hypofunction leads to molecular and 3D morphometric signs of atrophy of the masseter and its agonist masticatory muscles in adult mice. Twenty-three 8-week-old male BALB/c mice received a single injection of BoNTA in the right masseter, whereas the left masseter received the same volume of saline solution (control side). Animals were euthanized at 2d, 7d, and 14d, and the masticatory muscles were analyzed for mRNA expression. Five heads were harvested at 14d, fixed, stained with a contrast-enhanced agent, and scanned using X-ray microtomography. The three-dimensional morphometric parameters (the volume and thickness) from muscles in situ were obtained. Atrogin-1/MAFbx, MuRF-1, and Myogenin mRNA gene expression were significantly increased at 2 and 7d for both the masseter and temporalis from the BoNTA side. For medial pterygoid, increased mRNA gene expression was found at 7d for Atrogin-1/MAFbx and at 2d-7d for Myogenin. Both the volume and thickness of the masseter, temporalis, and medial pterygoid muscles from the BoNTA side were significantly reduced at 14d. In contrast, the lateral pterygoid from the BoNTA side showed a significant increase in volume at 14d. Therefore, the unilateral hypofunction of the masseter leads to molecular and morphological signs of atrophy in both the BoNTA-injected muscle and its agonistic non-injected masticatory muscles. The generalized effect on the mouse masticatory apparatus when one of its components is intervened suggests the need for more clinical studies to determine the safety of BoNTA usage in clinical dentistry.

Boldine modulates glial transcription and functional recovery in a murine model of contusion spinal cord injury.

Membrane channels such as those formed by connexins (Cx) and P2X7 receptors (P2X7R) are permeable to calcium ions and other small molecules such as adenosine triphosphate (ATP) and glutamate. Release of ATP and glutamate through these channels is a key mechanism driving tissue response to traumas such as spinal cord injury (SCI). Boldine, an alkaloid isolated from the Chilean boldo tree, blocks both Cx and Panx1 hemichannels (HCs). To test if boldine could improve function after SCI, boldine or vehicle was administered to treat mice with a moderate severity contusion-induced SCI. Boldine led to greater spared white matter and increased locomotor function as determined by the Basso Mouse Scale and horizontal ladder rung walk tests. Boldine treatment reduced immunostaining for markers of activated microglia (Iba1) and astrocytic (GFAP) markers while increasing that for axon growth and neuroplasticity (GAP-43). Cell culture studies demonstrated that boldine blocked glial HC, specifically Cx26 and Cx30, in cultured astrocytes and blocked calcium entry through activated P2X7R. RT-qPCR studies showed that boldine treatment reduced expression of the chemokine Ccl2, cytokine IL-6 and microglial gene CD68, while increasing expression of the neurotransmission genes Snap25 and Grin2b, and Gap-43. Bulk RNA sequencing revealed that boldine modulated a large number of genes involved in neurotransmission in spinal cord tissue just caudal from the lesion epicenter at 14 days after SCI. Numbers of genes regulated by boldine was much lower at 28 days after injury. These results indicate that boldine treatment ameliorates injury and spares tissue to increase locomotor function.

Skeletal Muscle Atrophy Induced by Diabetes Is Mediated by Non-Selective Channels and Prevented by Boldine.

Individuals with diabetes mellitus present a skeletal muscle myopathy characterized by atrophy. However, the mechanism underlying this muscular alteration remains elusive, which makes it difficult to design a rational treatment that could avoid the negative consequences in muscles due to diabetes. In the present work, the atrophy of skeletal myofibers from streptozotocin-induced diabetic rats was prevented with boldine, suggesting that non-selective channels inhibited by this alkaloid are involved in this process, as has previously shown for other muscular pathologies. Accordingly, we found a relevant increase in sarcolemma permeability of skeletal myofibers of diabetic animals in vivo and in vitro due to de novo expression of functional connexin hemichannels (Cx HCs) containing connexins (Cxs) 39, 43, and 45. These cells also expressed P2X7 receptors, and their inhibition in vitro drastically reduced sarcolemma permeability, suggesting their participation in the activation of Cx HCs. Notably, sarcolemma permeability of skeletal myofibers was prevented by boldine treatment that blocks Cx43 and Cx45 HCs, and now we demonstrated that it also blocks P2X7 receptors. In addition, the skeletal muscle alterations described above were not observed in diabetic mice with myofibers deficient in Cx43/Cx45 expression. Moreover, murine myofibers cultured for 24 h in high glucose presented a drastic increase in sarcolemma permeability and levels of NLRP3, a molecular member of the inflammasome, a response that was also prevented by boldine, suggesting that, in addition to the systemic inflammatory response found in diabetes, high glucose can promote the expression of functional Cx HCs and activation of the inflammasome in skeletal myofibers. Therefore, Cx43 and Cx45 HCs play a critical role in myofiber degeneration, and boldine could be considered a potential therapeutic agent to treat muscular complications due to diabetes.

Boldine modulates glial transcription and functional recovery in a murine model of contusion spinal cord injury.

Membrane channels such as connexins (Cx), pannexins (Panx) and P2X 7 receptors (P2X 7 R) are permeable to calcium ions and other small molecules such as ATP and glutamate. Release of ATP and glutamate through these channels is a key mechanism driving tissue response to traumas such as spinal cord injury (SCI). Boldine, an alkaloid isolated from the Chilean boldo tree, blocks both Cx hemichannels (HC) and Panx. To test if boldine could improve function after SCI, boldine or vehicle was administered to treat mice with a moderate severity contusion-induced SCI. Boldine led to greater spared white matter and increased locomotor function as determined by the Basso Mouse Scale and horizontal ladder rung walk tests. Boldine treatment reduced immunostaining for markers of activated microglia (Iba1) and astrocytic (GFAP) markers while increasing that for axon growth and neuroplasticity (GAP-43). Cell culture studies demonstrated that boldine blocked glial HC, specifically Cx26 and Cx30, in cultured astrocytes and blocked calcium entry through activated P2X 7 R. RT-qPCR studies showed that boldine treatment reduced expression of the chemokine Ccl2, cytokine IL-6 and microglial gene CD68, while increasing expression of the neurotransmission genes Snap25 and Grin2b, and Gap-43. Bulk RNA sequencing (of the spinal cord revealed that boldine modulated a large number of genes involved in neurotransmission in in spinal cord tissue just below the lesion epicenter at 14 days after SCI. Numbers of genes regulated by boldine was much lower at 28 days after injury. These results indicate that boldine treatment ameliorates injury and spares tissue to increase locomotor function.

Oxidative Stress, Inflammation and Connexin Hemichannels in Muscular Dystrophies.

Muscular dystrophies (MDs) are a heterogeneous group of congenital neuromuscular disorders whose clinical signs include myalgia, skeletal muscle weakness, hypotonia, and atrophy that leads to progressive muscle disability and loss of ambulation. MDs can also affect cardiac and respiratory muscles, impairing life-expectancy. MDs in clude Duchenne muscular dystrophy, Emery-Dreifuss muscular dystrophy, facioscapulohumeral muscular dystrophy and limb-girdle muscular dystrophy. These and other MDs are caused by mutations in genes that encode proteins responsible for the structure and function of skeletal muscles, such as components of the dystrophin-glycoprotein-complex that connect the sarcomeric-actin with the extracellular matrix, allowing contractile force transmission and providing stability during muscle contraction. Consequently, in dystrophic conditions in which such proteins are affected, muscle integrity is disrupted, leading to local inflammatory responses, oxidative stress, Ca2+-dyshomeostasis and muscle degeneration. In this scenario, dysregulation of connexin hemichannels seem to be an early disruptor of the homeostasis that further plays a relevant role in these processes. The interaction between all these elements constitutes a positive feedback loop that contributes to the worsening of the diseases. Thus, we discuss here the interplay between inflammation, oxidative stress and connexin hemichannels in the progression of MDs and their potential as therapeutic targets.

Muscle-Bone Crosstalk in the Masticatory System: From Biomechanical to Molecular Interactions.

The masticatory system is a complex and highly organized group of structures, including craniofacial bones (maxillae and mandible), muscles, teeth, joints, and neurovascular elements. While the musculoskeletal structures of the head and neck are known to have a different embryonic origin, morphology, biomechanical demands, and biochemical characteristics than the trunk and limbs, their particular molecular basis and cell biology have been much less explored. In the last decade, the concept of muscle-bone crosstalk has emerged, comprising both the loads generated during muscle contraction and a biochemical component through soluble molecules. Bone cells embedded in the mineralized tissue respond to the biomechanical input by releasing molecular factors that impact the homeostasis of the attaching skeletal muscle. In the same way, muscle-derived factors act as soluble signals that modulate the remodeling process of the underlying bones. This concept of muscle-bone crosstalk at a molecular level is particularly interesting in the mandible, due to its tight anatomical relationship with one of the biggest and strongest masticatory muscles, the masseter. However, despite the close physical and physiological interaction of both tissues for proper functioning, this topic has been poorly addressed. Here we present one of the most detailed reviews of the literature to date regarding the biomechanical and biochemical interaction between muscles and bones of the masticatory system, both during development and in physiological or pathological remodeling processes. Evidence related to how masticatory function shapes the craniofacial bones is discussed, and a proposal presented that the masticatory muscles and craniofacial bones serve as secretory tissues. We furthermore discuss our current findings of myokines-release from masseter muscle in physiological conditions, during functional adaptation or pathology, and their putative role as bone-modulators in the craniofacial system. Finally, we address the physiological implications of the crosstalk between muscles and bones in the masticatory system, analyzing pathologies or clinical procedures in which the alteration of one of them affects the homeostasis of the other. Unveiling the mechanisms of muscle-bone crosstalk in the masticatory system opens broad possibilities for understanding and treating temporomandibular disorders, which severely impair the quality of life, with a high cost for diagnosis and management.

Niña con erucismo hemorrágico por Lonomia spp.: / Hemorrhagic erucism due to Lonomia spp. in a girl:

RESUMEN Los accidentes causados por las setas urticantes o venenosas de las orugas de lepidópteros, se conocen como erucismo. Estos accidentes se producen por el contacto accidental, especialmente por los niños, con las cerdas sobre el cuerpo del insecto, conectadas con glándulas venenosas. Los síntomas pueden ser locales o sistémicos, con presentaciones clínicas fatales. El accidente ocasionado por las orugas del género Lonomia spp. puede desencadenar síndromes hemorrágicos, constituyendo estos la forma más grave de erucismo. Se reporta el caso de una niña de 5 años, procedente del poblado de Villarondos, en la Amazonía del Perú, departamento de Huánuco, la cual incidentalmente se hinca con las cerdas de una oruga, cursando luego con anemia hemolítica, plaquetopenia y trastorno de la coagulación. El diagnóstico fue establecido por la anamnesis, cuadro clínico, exámenes de laboratorio y respuesta al suero antilonómico. Se discuten los aspectos clínicos, laboratoriales y terapéuticos de erucismo por Lonomia spp.

ABSTRACT Accidents caused by urticating or poisonous setae from lepidoptera caterpillars are known as erucism. These accidents are produced by accidental contact, especially in children, with bristles on the insect's body surface, connected to venom glands. Symptoms may be local or systemic, with deadly clinical presentations. The accident caused by Lonomia spp. caterpillars can trigger bleeding disorders, which is considered the most severe type of erucism. The case of a 5-year-old girl is reported. She was from the town of Villarondos, in the Peruvian Amazon, department of Huánuco, who accidentally knelt down on caterpillar bristles, and subsequently experienced hemolytic anemia, plateletopenia, and coagulation disorder. The diagnosis was made based on the medical history, clinical manifestation, laboratory examination results, and response to antilonomic serum. The clinical, laboratory, and therapeutic aspects of erucism due to Lonomia spp. are discussed.

[Hemorrhagic erucism due to Lonomia spp. in a girl]. / Niña con erucismo hemorrágico por Lonomia spp.

Accidents caused by urticating or poisonous setae from lepidoptera caterpillars are known as erucism. These accidents are produced by accidental contact, especially in children, with bristles on the insect's body surface, connected to venom glands. Symptoms may be local or systemic, with deadly clinical presentations. The accident caused by Lonomia spp. caterpillars can trigger bleeding disorders, which is considered the most severe type of erucism. The case of a 5-year-old girl is reported. She was from the town of Villarondos, in the Peruvian Amazon, department of Huánuco, who accidentally knelt down on caterpillar bristles, and subsequently experienced hemolytic anemia, plateletopenia, and coagulation disorder. The diagnosis was made based on the medical history, clinical manifestation, laboratory examination results, and response to antilonomic serum. The clinical, laboratory, and therapeutic aspects of erucism due to Lonomia spp. are discussed.

Evaluación de la protección conferida por un programa de vacunación contra la enfermedad de Gumboro en pollos de carne aplicando la fórmula Deventer / Evaluation of the level of protection conferred by a vaccination scheme against the infectious bursal disease in broilers using the Deventer

El presente estudio evaluó la protección conferida por un programa de vacunación a la edad determinada por la fórmula Deventer frente a un desafío experimental con una cepa estándar del virus de la Enfermedad Infecciosa de la Bursa (EIB) en pollos de carne. Se formaron tres grupos experimentales. El grupo A vacunado a los 10 días de edad con la cepa Lukert (intermedia-suave) y a los 18 días con la cepa CE (intermedia-intermedia); el grupo B vacunado a los 21 días de edad (según la fórmula Deventer) con la cepa 228TC (intermedia-intermedia); y el grupo C no vacunado (control). A los 35 días de edad, 45 aves de cada grupo fueron desafiadas con la cepa estándar F 52/70 del virus de la EIB. La protección fue medida a través de signos clínicos, índice bursal (I.B.), lesiones macroscópicas y microscópicas, y serología. Todos los grupos presentaron signos clínicos y edema bursal hasta el día 10 post desafío. Los valores de I.B. en los tres grupos fueron compatibles con atrofia bursal y las lesiones histopatológicas fueron severas en los grupos vacunados. No se observó seroconversión a EIB hasta el final del estudio. Los resultados obtenidos indicaron que ninguno de los grupos fue protegido de la enfermedad clínica hasta los 10 días post desafío. Sin embargo, la mejor protección se observó en el grupo A en comparación al grupo B, aunque sin encontrar diferencias estadísticas.

The present study evaluated the conferred protection by a program of vaccination at an age that was determined by the Deventer formula against a challenge with a standard strain of the Infectious Bursal Disease virus (IBDV) in broiler chickens. There were three experimental groups. The group A was vaccinated at 10 days of age with a Lukert strain (intermediate-mild) and at 18 days with CE strain (intermediate-intermediate); group B vaccinated at 21 days old (according to the Deventer formula) with the 228TC strain (intermediate-intermediate); and group C were not vaccinated (control). At 35 days of age, 45 birds of each group were challenged with a standard strain F 52/70 of the IBDV. The protection was measured through clinical signs, bursal index (B.I.), gross and microscopic lesions, and serology. All groups presented clinical signs and bursal oedema until 10 days post challenge. The values of B.I. in the three groups were compatible with bursal atrophy and the histopathology lesions were severe in the vaccinated groups. Seroconversion was not observed until the end of the study. The obtained results indicated that birds in none of the experimental groups were protected against the clinical disease until 10 days post challenge. The best protection was observed in the group A in comparison group B but without statistical difference.

Indicaciones y colocación de catéteres venosos centrales en situaciones críticas / Indications and complications of central venous cannulation in critical patients

Se señalan las vías de abordaje más adecuadas, sus indicaciones y complementaciones en la colocación de Catéteres Venosos Centrales en pacientes que se encuentran en situaciones críticas en la Sala de Emergencia de un Hospital General. Se colocaron 145 CVC en 144 pacientes, los que, con excepción de 02 pacientes, tuvieron control Radiológico. La principal indicación para la colocación del CVC fue la determinación de la PVC (86 por ciento) y la dificultad para conseguir una vía adecuada (14 por ciento). Las principales vías de abordaje fueron: VSCD/SC (77 por ciento), VYID (18 por ciento), VYII(4 por ciento), VSCI/SC (2 por ciento). Hubieron fallas en el 5 por ciento de las colocaciones del CVC, siendo significativo en la VYI, 12 pacientes presentaron complicaciones (8 por ciento), siendo lo más frecuente hematomas en el abordaje de la VYI,1 paciente presentó neumotórax masivo en el abordaje de la VSCD. La colocación de un CVC es de extrema utilidad en pacientes críticos, su uso requiere de experiencia y de una técnica adecuada. En el presente estudio el abordaje de la VSC/SC resultó siendo el más sencillo y demandó menor tiempo en la Sala de Emergencia, sin embargo el abordaje de la VYI resultó siendo la más segura al no tener complicaciones mayores.