Serum Neurofilament Light-Chain Concentrations in Essential Tremor: a Case-Control Study.

Essential tremor (ET) is a common neurological disorder, with clinical and pathophysiological links to the cerebellum. Inquiries into the etiology, pathophysiology, and nosology of ET stand to benefit from the identification of disease biomarkers. Serum neurofilament light chain (NfL) has emerged as a novel signature of conditions in which neuronal injury reflects an outcome of the ongoing disease process. We sought to investigate the concentrations of NfL in ET patients and healthy controls. In this case-control study, our powered study population of 41 ET patients and 40 age-matched healthy controls underwent clinical assessments and measurement of serum NfL concentration using Simoa technology. Serum NfL was elevated in ET patients - mean log-transformed serum NfL concentration = 1.23 ± 0.19 (95% confidence interval [CI] = 1.17-1.29) vs. 1.08 ± 0.15 (95% CI = 1.03-1.13), p = 0.0002. This difference persisted after accounting for age, sex and Montreal Cognitive Assessment score in a multiple linear regression model (p = 0.002) and in an age-matched sample subset of 35 ET cases and 35 controls (p = 0.006). There was no association between tremor severity and serum NfL levels (p = 0.73). In this sample of ET patients and controls, serum NfL concentrations were significantly higher in ET. Studies in additional cohorts of ET cases would be of value in attempting to replicate these results and assessing diagnostic utility.

Primer Consenso Nacional Intersociedades de Inmunización del Adulto. 2023

Las vacunas previenen millones de muertes cada año y su eficacia y seguridad han sido ampliamente establecidas. En términos económicos, la vacunación es una de las intervenciones sanitarias más costo efectivas, generando un importante ahorro y crecimiento económico que supone a largo plazo. Se ha demostrado que la vacunación de adultos disminuye la morbilidad y la mortalidad asociadas a enfermedades infecciosas prevenibles, reduciendo las complicaciones y las hospitalizaciones, incluidos los ingresos a las unidades de cuidados intensivos. Hemos elaborado este documento de consenso con el objeto de diseñar un esquema de vacunación pragmático, accesible y estandarizado del adulto, según categoría de riesgo y edad, sobre la base de la evidencia disponible de vacunas accesibles y nuevas vacunas habiendo utilizado el Tercer Consenso de la Sociedad Paraguaya de Infectología del 2019 como base para las recomendaciones finales.

SUMMARY Vaccines prevent millions of deaths each year, and their efficacy and safety have been widely established. In economic terms, vaccination is one of the most cost-effective health interventions, generating significant savings and long-term economic growth. Adult vaccination has been shown to decrease morbidity and mortality associated with preventable Infectious diseases, reducing complications and hospitalizations, including admissions to intensive care units. We have prepared this consensus document in order to design a pragmatic, accessible and standardized vaccination scheme for adults, according to risk category and age, based on the available evidence of available vaccines and new vaccines, having used the third consensus of the Paraguayan Infectious Diseases Society of 2019 as a basis for the final recommendations.

Distinctive transcriptomic and epigenomic signatures of bone marrow-derived myeloid cells and microglia in CNS autoimmunity.

In the context of autoimmunity, myeloid cells of the central nervous system (CNS) constitute an ontogenically heterogeneous population that includes yolk sac-derived microglia and infiltrating bone marrow-derived cells (BMC). We previously identified a myeloid cell subset in the brain and spinal cord that expresses the surface markers CD88 and CD317 and is associated with the onset and persistence of clinical disease in the murine model of the human CNS autoimmune disorder, experimental autoimmune encephalomyelitis (EAE). We employed an experimental platform utilizing single-cell transcriptomic and epigenomic profiling of bone marrow-chimeric mice to categorically distinguish BMC from microglia during CNS autoimmunity. Analysis of gene expression and chromosomal accessibility identified CD88+CD317+ myeloid cells in the CNS of EAE mice as originating from BMC and microglia. Interestingly, each cell lineage exhibited overlapping and unique gene expression patterns and transcription factor motifs that allowed their segregation. Our observations will facilitate determining pathogenic contributions of BMC and microglia in CNS autoimmune disease. Ultimately, this agnostic characterization of myeloid cells will be required for devising disease stage-specific and tissue-specific interventions for CNS inflammatory and neurodegenerative disorders.

Assessment of Regional Lung Ventilation with Positron Emission Tomography Using the Radiofluorinated Gas [18F]SF6: Application to an Animal Model of Impaired Ventilation.

PURPOSE: Clinical ventilation studies are primarily performed with computerized tomography (CT) and more often with single-photon emission Computerized tomography (SPECT) using radiolabelled aerosols, both presenting certain limitations. Here, we investigate the use of the radiofluorinated gas [18F]SF6 as a positron emission tomography (PET) ventilation marker in an animal model of impaired lung ventilation. PROCEDURES: Sprague-Dawley rats (n = 15) were randomly assigned to spontaneous ventilation (sham group), endotracheal administration of phosphate-buffered saline (PBS group), or endotracheal administration of lipopolysaccharide (LPS group). PET-[18F]SF6 images (10-min acquisition) were acquired at t = 48 h after LPS or PBS administration under mechanical ventilation. CT images were acquired after each PET session. Volumes of interest were manually delineated in the lungs on CT images, and voxel-by-voxel analysis was carried out on PET images to obtain the corresponding histograms. After the imaging sessions, lungs were harvested to conduct histological analysis. RESULTS: Ventilation studies in sham animals showed uniform distribution of [18F]SF6 and fast elimination of the radioactivity after discontinuation of the administration. For PBS- and LPS-treated rats, ventilation defects were observed on PET images in some animals, identified as regions with low presence of the radiolabelled gas. Hypoventilated areas co-localized with regions with higher x-ray attenuation than healthy lungs on the CT images, suggesting the presence of oedema and, in some cases, atelectasis. Histograms obtained from PET images showed quasi-Gaussian distributions for control animals, while PBS- and LPS-treated animals demonstrated the presence of hypoventilated voxels. Deviation of the histograms from Gaussian distribution correlated with histological score was obtained by ex vivo histological analysis. CONCLUSIONS: [18F]SF6 is an appropriate marker of regional lung ventilation and may find application in the early diagnose of acute lung disease.

Clinical trials in multiple sclerosis: past, present, and future.

For the past four decades, multiple sclerosis (MS) has been a focus for clinical trial development and execution. Advances in translational neuroimmunology have led to the development of effective disease-modifying therapies (DMTs) that greatly benefit patients with MS and mitigate their burden of disease. These achievements also stem from continued progress made in the definition and discovery of sensitive disease diagnostic criteria, objective disability assessment scales, precise imaging techniques, and disease-specific biomarkers. As a result, our knowledge of MS pathophysiology is more mature; the established clinical practice for the diagnosis and management of MS could serve as a roadmap to guide the development of more disease-specific interventions. In this article we briefly review the main achievements in the evolution of clinical trials for MS, and discuss opportunities for improvements.

Efficacy of Disease Modifying Therapies in Progressive MS and How Immune Senescence May Explain Their Failure.

The advent of disease modifying therapies (DMT) in the past two decades has been the cornerstone of successful clinical management of multiple sclerosis (MS). Despite the great strides made in reducing the relapse frequency and occurrence of new signal changes on neuroimaging in patients with relapsing remitting MS (RRMS) by approved DMT, it has been challenging to demonstrate their effectiveness in non-active secondary progressive MS (SPMS) and primary progressive MS (PPMS) disease phenotypes. The dichotomy of DMT effectiveness between RRMS and progressive MS informs on distinct pathogeneses of the different MS phenotypes. Conversely, factors that render patients with progressive MS resistant to therapy are not understood. Thus far, age has emerged as the main correlate of the transition from RRMS to SPMS. Whether it is aging and age-related factors or the underlying immune senescence that qualitatively alter immune responses as the disease transitions to SPMS, that diminish DMT effectiveness, or both, is currently not known. Here, we will discuss the role of immune senescence on different arms of the immune system, and how it may explain relative DMT resistance.

Histiocitosis de células de langerhans. Un hallazgo poco común en la glándula tiroides. A propósito de un caso / Langerhans cell histiocytosis. A rare finding in the thyroid gland. About a case

La histiocitosis de células de Langerhans (HCL) es una neoplasia mieloide, poco común, se caracteriza por la proliferación de células histiocíticas CD1a+/Langerina+. La incidencia en adultoses1 a 2 casos por millón,unpoco más frecuente en niños. La presentación es variable, como una única lesión en un órgano hasta una enfermedad multisistémica con toque de órganos vitales. Los tejidos de mayor afinidad son: huesos, piel, pulmones e hipófisis y raramente tiroides.El retraso en el diagnóstico o en el tratamiento puede provocar un deterioro de la función de los órganos afectados. El objetivo de la comunicación es reportar un caso de HCL, de presentación aún menos frecuente, en forma aislada en la glándula tiroides de un adulto, hallazgo hecho tras una tiroidectomía total, realizándose el diagnóstico luego de la histología.

Langerhans cell histiocytosis (LCH) is a rare myeloid neoplasm characterized by the proliferation of CD1a+/Langerin+ histiocytic cells. The incidence in adultsis1 to 2 cases per million, a little more frequent in children. The presentationis variable, as a single lesion in an organ to a multisystemic disease with touch of vital organs. The tissues with the highest affinity are: bones, skin, lungs and pituitary and rarely thyroid. Delay in the diagnosis or in the treatment can cause a deterioration in thefunction of the affected organs. The objective of the communication is to report a case of LCH, with an even less frequent presentation, isolated formin the thyroid gland of an adult, a finding made after a total thyroidectomy, the diagnosis being made after histology.

Systems Approaches to Unravel T Cell Function and Therapeutic Potential in Autoimmune Disease.

Producing Ag-specific immune responses constrained to target tissues or cells that can be engaged or disengaged at will is predicated on understanding the network of genes governing immune cell function, defining the rules underlying Ag specificity, and synthesizing the tools to engineer them. The successes and limitations of chimeric Ag receptor (CAR) T cells emphasize this goal, and advances in high-throughput sequencing, large-scale genomic screens, single-cell profiling, and genetic modification are providing the necessary data to bring it to fruition-including a broader application into the treatment of autoimmune diseases. In this review, we delve into the implementation of these developments, survey the relevant works, and propose a framework for generating the next generation of synthetic T cells informed by the principles learned from these systems approaches.

Evidence of a landlocked reproducing population of the marine pejerrey Odontesthes argentinensis (Actinopterygii; Atherinopsidae).

In South America, the order Atheriniformes includes the monophyletic genus Odontesthes with 20 species that inhabit freshwater, estuarine and coastal environments. Pejerrey Odontesthes argentinensis is widely distributed in coastal and estuarine areas of the Atlantic Ocean and is known to foray into estuaries of river systems, particularly in conditions of elevated salinity. However, to our knowledge, a landlocked self-sustaining population has never been recorded. In this study, we examined the pejerrey population of Salada de Pedro Luro Lake (south-east of Buenos Aires Province, Argentina) to clarify its taxonomic identity. An integrative taxonomic analysis based on traditional meristic, landmark-based morphometrics and genetic techniques suggests that the Salada de Pedro Luro pejerrey population represents a novel case of physiological and morphological adaptation of a marine pejerrey species to a landlocked environment and emphasises the environmental plasticity of this group of fishes.

Coevolution-based inference of amino acid interactions underlying protein function.

Protein function arises from a poorly understood pattern of energetic interactions between amino acid residues. Sequence-based strategies for deducing this pattern have been proposed, but lack of benchmark data has limited experimental verification. Here, we extend deep-mutation technologies to enable measurement of many thousands of pairwise amino acid couplings in several homologs of a protein family - a deep coupling scan (DCS). The data show that cooperative interactions between residues are loaded in a sparse, evolutionarily conserved, spatially contiguous network of amino acids. The pattern of amino acid coupling is quantitatively captured in the coevolution of amino acid positions, especially as indicated by the statistical coupling analysis (SCA), providing experimental confirmation of the key tenets of this method. This work exposes the collective nature of physical constraints on protein function and clarifies its link with sequence analysis, enabling a general practical approach for understanding the structural basis for protein function.

First molecular data for Lernaea cyprinacea (Copepoda: Cyclopoida) infesting Odontesthes bonariensis, a commercially important freshwater fish in Argentina.

Parasitic copepods of the family Lernaeidae are often found infesting freshwater fishes worldwide. They cause lernaeosis, a disease that can lead to serious pathogenic effects on their fish hosts. The most common lernaeid is the Lernaea cyprinacea, which has been widely introduced through importation of tropical fishes, e.g. cyprinids. In South America, it is one of the most common parasites both in wild and in farmed fish in the central region of Argentina. The silverside Odontesthes bonariensis is the most important fish of the sport fisheries of Argentina and one of the fish most affected by lernaeosis. Six specimens of copepods were collected from 30 specimens of O. bonariensis collected in a Pampean shallow lake (33°25'28"S 62°53'56"W) of Córdoba (Argentina). The 28S rRNA gene of L. cyprinacea was amplified by means of PCR to obtain the 28S rDNA sequence. The sequence obtained of this parasite from Argentina showed high genetic similarity with those from various geographical origins. The present study provided molecular characterization of L. cyprinacea in South America for the first time.

First molecular data for Lernaea cyprinacea (Copepoda: Cyclopoida) infesting Odontesthes bonariensis, a commercially important freshwater fish in Argentina / Primeiros dados moleculares para Lernaea cyprinacea (Copepoda: Cyclopoida) infestando Odontesthes bonariensis, um peixe de água doce comercialmente importante na Argentina

Abstract Parasitic copepods of the family Lernaeidae are often found infesting freshwater fishes worldwide. They cause lernaeosis, a disease that can lead to serious pathogenic effects on their fish hosts. The most common lernaeid is the Lernaea cyprinacea, which has been widely introduced through importation of tropical fishes, e.g. cyprinids. In South America, it is one of the most common parasites both in wild and in farmed fish in the central region of Argentina. The silverside Odontesthes bonariensis is the most important fish of the sport fisheries of Argentina and one of the fish most affected by lernaeosis. Six specimens of copepods were collected from 30 specimens of O. bonariensis collected in a Pampean shallow lake (33°25'28"S 62°53'56"W) of Córdoba (Argentina). The 28S rRNA gene of L. cyprinacea was amplified by means of PCR to obtain the 28S rDNA sequence. The sequence obtained of this parasite from Argentina showed high genetic similarity with those from various geographical origins. The present study provided molecular characterization of L. cyprinacea in South America for the first time.

Resumo Os copépodos parasitos da família Lernaeidae são frequentemente encontrados infestando peixes de água doce em todo o mundo, causando a lernaeose, uma doença que pode levar a graves efeitos patogênicos em seus hospedeiros. O lerneídeo mais comum é a Lernaea cyprinacea, que tem sido amplamente introduzida por meio da importação de peixes tropicais, tais como ciprinídeos. Na América do Sul, é um dos parasitos mais comuns em peixes selvagens, bem como em peixes de cultivo na região central argentina. Odontesthes bonariensis é um dos peixes mais importantes para a pesca esportiva na Argentina e um dos mais afetados pela lerneose. Um total de seis espécimes desses copépodes foram coletados de espécimes de 30 O. bonariensis capturados em um lago de pouca profundidade nos Pampas (33°25'28"S 62°53'56"W) em Córdoba (Argentina). O gene 28S rRNA de L. cyprinacea foi amplificado por PCR para obter a sequência do 28S rDNA. A sequência obtida desse parasito da Argentina mostrou alta similaridade genética com aquelas de outras origens geográficas. O presente estudo forneceu, pela primeira vez, uma caracterização molecular de L. cyprinacea na América do Sul.

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing.

Site-directed mutagenesis has long been used as a method to interrogate protein structure, function and evolution. Recent advances in massively-parallel sequencing technology have opened up the possibility of assessing the functional or fitness effects of large numbers of mutations simultaneously. Here, we present a protocol for experimentally determining the effects of all possible single amino acid mutations in a protein of interest utilizing high-throughput sequencing technology, using the 263 amino acid antibiotic resistance enzyme TEM-1 ß-lactamase as an example. In this approach, a whole-protein saturation mutagenesis library is constructed by site-directed mutagenic PCR, randomizing each position individually to all possible amino acids. The library is then transformed into bacteria, and selected for the ability to confer resistance to ß-lactam antibiotics. The fitness effect of each mutation is then determined by deep sequencing of the library before and after selection. Importantly, this protocol introduces methods which maximize sequencing read depth and permit the simultaneous selection of the entire mutation library, by mixing adjacent positions into groups of length accommodated by high-throughput sequencing read length and utilizing orthogonal primers to barcode each group. Representative results using this protocol are provided by assessing the fitness effects of all single amino acid mutations in TEM-1 at a clinically relevant dosage of ampicillin. The method should be easily extendable to other proteins for which a high-throughput selection assay is in place.

Binding and Endocytosis of Bovine Hololactoferrin by the Parasite Entamoeba histolytica.

Entamoeba histolytica is a human parasite that requires iron (Fe) for its metabolic function and virulence. Bovine lactoferrin (B-Lf) and its peptides can be found in the digestive tract after dairy products are ingested. The aim of this study was to compare virulent trophozoites recently isolated from hamster liver abscesses with nonvirulent trophozoites maintained for more than 30 years in cultures in vitro regarding their interaction with iron-charged B-Lf (B-holo-Lf). We performed growth kinetics analyses of trophozoites in B-holo-Lf and throughout several consecutive transfers. The virulent parasites showed higher growth and tolerance to iron than nonvirulent parasites. Both amoeba variants specifically bound B-holo-Lf with a similar K d . However, averages of 9.45 × 10(5) and 6.65 × 10(6) binding sites/cell were found for B-holo-Lf in nonvirulent and virulent amoebae, respectively. Virulent amoebae bound more efficiently to human and bovine holo-Lf, human holo-transferrin, and human and bovine hemoglobin than nonvirulent amoebae. Virulent amoebae showed two types of B-holo-Lf binding proteins. Although both amoebae endocytosed this glycoprotein through clathrin-coated vesicles, the virulent amoebae also endocytosed B-holo-Lf through a cholesterol-dependent mechanism. Both amoeba variants secreted cysteine proteases cleaving B-holo-Lf. These data demonstrate that the B-Lf endocytosis is more efficient in virulent amoebae.

PD-L1 regulates the development, maintenance, and function of induced regulatory T cells.

Both the programmed death (PD) 1-PD-ligand (PD-L) pathway and regulatory T (T reg) cells are instrumental to the maintenance of peripheral tolerance. We demonstrate that PD-L1 has a pivotal role in regulating induced T reg (iT reg) cell development and sustaining iT reg cell function. PD-L1(-/-) antigen-presenting cells minimally convert naive CD4 T cells to iT reg cells, showing the essential role of PD-L1 for iT reg cell induction. PD-L1-coated beads induce iT reg cells in vitro, indicating that PD-L1 itself regulates iT reg cell development. Furthermore, PD-L1 enhances and sustains Foxp3 expression and the suppressive function of iT reg cells. The obligatory role for PD-L1 in controlling iT reg cell development and function in vivo is illustrated by a marked reduction in iT reg cell conversion and rapid onset of a fatal inflammatory phenotype in PD-L1(-/-)PD-L2(-/-) Rag(-/-) recipients of naive CD4 T cells. PD-L1 iT reg cell development is mediated through the down-regulation of phospho-Akt, mTOR, S6, and ERK2 and concomitant with the up-regulation of PTEN, all key signaling molecules which are critical for iT reg cell development. Thus, PD-L1 can inhibit T cell responses by promoting both the induction and maintenance of iT reg cells. These studies define a novel mechanism for iT reg cell development and function, as well as a new strategy for controlling T reg cell plasticity.

Biología del pejerrey Odontesthes bonariensis (Pisces, Atherinopsidae) de la laguna Los Charos (Córdoba, Argentina) / Biology of silverside Odontesthes bonariensis (Pisces, Atherinopsidae) in the shallow lake “Los Charos” (Córdoba, Argentina)

El pejerrey Odontesthes bonariensis es la especie más importante de las pesquerías del centro de Argentina. Se estudio la captura por unidad de esfuerzo (CPUE), la condición corporal, el crecimiento y la alimentación de O. bonariensis de la laguna pampeana Los Charos (34º28´S, 64º23´W, 240 ha), ubicada en la provincia de Córdoba. Se realizaron cuatro muestreos estacionales en el periodo 2002-2003. Para la captura de peces se utilizaron redes de arrastre y enmalle. Se capturaron 2862 ejemplares de un rango de talla comprendido entre 38 y 380 mm de longitud estándar (LSt). La CPUE promedio fue de 74,3 (±71,0)kg/20 hs de tendido de red. La relación LSt-peso presentó diferencias significativas entre épocas del año (P< 0,01). Los índices de condición corporal estuvieron dentro de los límites de referencia de la especie. El crecimiento calculado fue: LSt(t)=459,8*[1–exp(-0,3105*(t-0,175))]. La relación LSt–Longitud total (LT) fue: LT(mm)=8,23+LSt*1,14 (n=283; R2=0,99). El zooplancton constituyó un ítem alimenticio secundario en los peces jóvenes. En los ejemplares de 3+ años de vida se observó un marcado canibalismo, situación que explicaría en parte su mejor condición corporal. La laguna Los Charos presenta una elevada producción de O. bonariensis.

The silverside Odontesthes bonariensis is the most important species in the fisheries of central Argentina. Capture per unit effort (CPUE), body condition, growth and feeding were studied for O. bonariensis in the Pampean shallow lake “Los Charos” (34º28´S, 64º23´W, 240 ha), located in Cordoba province. Four seasonal sampling procedures were carried out during 2002-2003. Floating gillnets and trawlnets were used to capture the fishes. Specimens captured (n= 2862) were ranged between 38 and 380 mm of standar length (StL). CPUE average was 74.3(±71,0) kg/20 hs/gillnets. The StL–weight relationship showed signifficant differences along the seasons (P< 0,01). The body score indices were within the reffered limits for this species. Growth was calculated by StL(t)= 459,8*[1–exp(-0,3105*(t-0,175))]. The StL–Total length (TL) ratio was TL(mm)= 8,23+StL*1,14 (n= 283; R2= 0,99). For the young fishes the zooplankton constituted a secundary feeding source. In individuals older than 3 years of age, a marked canibalism was observed, which might explain partially their body condition. The shallow lake “Los Charos” shows an elevated production of O. bonariensis.

Seasonal variation of parasitism in pejerey Odontesthes bonariensis (Atheriniformes, Atherinopsidae) from La Viña reservoir (Córdoba, Argentina).

The pejerrey Odontesthes bonariensis is the most important freshwater fish of Argentina central region. The present study was conducted to describe the seasonal variation of parasitism in O. bonariensis from La Viña reservoir (31 degrees 17'S, 65 degrees 01'W), located in the province of Córdoba, Argentina. Between years 1999 and 2002, 573 fishes were caught, and their parasite infection and relative weight (Wr) were assesed. Three parasites were found, Lernaea cyprinacea (Crustacea), Contracaecum spp. larvae (Nematoda) and Cangatiella macdonaghi (Cestoda). Overall, Contracaecum spp. prevalence and intensity were very low. Lernaea cyprinacea and Cangatiella macdonaghi prevalences increased during summer and spring (P < 0.01), respectively. Lernaea cyprinacea infection showed also seasonal differences in intensity (P < 0.05), as well as C. macdonaghi regarding parasites density (P < 0.01). Cangatiella macdonaghi average parasites density was 32.0 (+/-22.5) cestodes per cm(3)of digestive content. No association between parasitism and fish relative weight was found. Besides the seasonality variations observed, the parasite burden found did not compromise fish body condition.

Activation of protein kinase A stimulates the progesterone-induced calcium influx in human sperm exposed to the phosphodiesterase inhibitor papaverine.

Progesterone induces a fast transient calcium influx in human sperm though the activation of nongenomic receptors. During sperm capacitation, a complex process required for sperm to be able to fertilize the egg, the calcium influx induced by progesterone is enhanced. Sperm capacitation is mediated by an increase in cAMP content and subsequent protein kinase A (PKA) activation. In this work, we examined the effect of increasing intracellular cAMP on the calcium influx induced by progesterone in noncapacitated human sperm. To do this, sperm were exposed to the phosphodiesterase inhibitor papaverine for 5 minutes, a treatment that increased both the cAMP content and the PKA activity several-fold. The calcium influx induced by progesterone was increased by papaverine to levels close to those found in capacitated sperm. This effect was partially inhibited by H89 (48%) and by genistein (41%), and the sum of both inhibitors reduced the stimulating effect of papaverine by 89%. The inhibitory effect of genistein on the progesterone-induced calcium influx could be related to its capability to inhibit the papaverine-stimulated increase in cAMP content and PKA activity. The results presented here suggest that the calcium influx induced by progesterone is up-regulated by the PKA activity.