Rho-Associated Protein Kinase Activity Is Required for Tissue Homeostasis in the Xenopus laevis Ciliated Epithelium.

Lung epithelial development relies on the proper balance of cell proliferation and differentiation to maintain homeostasis. When this balance is disturbed, it can lead to diseases like cancer, where cells undergo hyperproliferation and then can undergo migration and metastasis. Lung cancer is one of the deadliest cancers, and even though there are a variety of therapeutic approaches, there are cases where treatment remains elusive. The rho-associated protein kinase (ROCK) has been thought to be an ideal molecular target due to its role in activating oncogenic signaling pathways. However, in a variety of cases, inhibition of ROCK has been shown to have the opposite outcome. Here, we show that ROCK inhibition with y-27632 causes abnormal epithelial tissue development in Xenopus laevis embryonic skin, which is an ideal model for studying lung cancer development. We found that treatment with y-27632 caused an increase in proliferation and the formation of ciliated epithelial outgrowths along the tail edge. Our results suggest that, in certain cases, ROCK inhibition can disturb tissue homeostasis. We anticipate that these findings could provide insight into possible mechanisms to overcome instances when ROCK inhibition results in heightened proliferation. Also, these findings are significant because y-27632 is a common pharmacological inhibitor used to study ROCK signaling, so it is important to know that in certain in vivo developmental models and conditions, this treatment can enhance proliferation rather than lead to cell cycle suppression.

Diosmetin attenuates fibromyalgia-like symptoms in a reserpine-induced model in mice.

Fibromyalgia is a potentially disabling idiopathic disease characterized by widespread chronic pain associated with comorbidities such as fatigue, anxiety, and depression. Current therapeutic approaches present adverse effects that limit adherence to therapy. Diosmetin, an aglycone of the flavonoid glycoside diosmin found in citrus fruits and the leaves of Olea europaea L., has antinociceptive, anti-inflammatory, and antioxidant properties. Here, we investigated the effect of diosmetin on nociceptive behaviors and comorbidities in an experimental fibromyalgia model induced by reserpine in mice. To induce the experimental fibromyalgia model, a protocol of subcutaneous injections of reserpine (1 mg/kg) was used once a day for three consecutive days in adult male Swiss mice. Mice received oral diosmetin on the fourth day after the first reserpine injection. Nociceptive (mechanical allodynia, muscle strength, and thermal hyperalgesia) and comorbid (depressive-like and anxiety behavior) parameters were evaluated. Potential adverse effects associated with diosmetin plus reserpine (locomotor alteration, cataleptic behavior, and body weight and temperature changes) were also evaluated. Oral diosmetin (0.015-1.5 mg/kg) reduced the mechanical allodynia, thermal hyperalgesia, and loss of muscle strength induced by reserpine. Diosmetin (0.15 mg/kg) also attenuated depressive-like and anxiety behaviors without causing locomotor alteration, cataleptic behavior, and alteration in weight and body temperature of mice. Overall, diosmetin can be an effective and safe therapeutic alternative to treat fibromyalgia symptoms, such as pain, depression and anxiety.

Effects of calcium supplementation on changes in the IL2, IL4, IL6, IL10 axes and oxidative stress in pregnant women at risk for pre-eclampsia.

BACKGROUND: Pregnant women with hypertensive disorders are at increased risk for inflammatory diseases and oxidative stress. The dilemma raised by the best dosage of calcium supplementation on these factors is evident. The aim of the current study was to examine the effects of calcium on biomarkers of the purinergic system, inflammation and oxidative stress, which are factors contributing to vascular damage in pregnant women at high risk of pre-eclampsia. METHODS: A prospective, double-blind and placebo-controlled study conducted with 101 women at risk of pre-eclampsia were randomized to take 500 mg calcium/day or 1,500 mg calcium/day or placebo for 6 weeks from the 20th gestational week until delivery. Fasting blood samples were collected at the beginning of the study and 6 weeks after the intervention. RESULTS: Taking calcium supplements (500 mg calcium/day) led to a significant increase in ATP hydrolysis (p < 0.05), NTPDase activity with increased hydrolysis of ADP and AMP nucleotides in platelets and lymphocytes. In the intragroup analysis IL-2, IL-6, IL-4 and interferon-É£ presented lower values in the calcium 1,500 mg/day group (p < 0.005). Oxidative stress was assessed by TBARS pro-oxidant marker, with an increase for the calcium groups when compared to the placebo group. The Vitamin C antioxidant marker presented a significant increase (p < 0.005) for the group that received high calcium doses. CONCLUSIONS: Calcium administration for 6 weeks had antioxidant action and positively modulated the purinergic system and inflammatory markers in pregnant women at risk of pre-eclampsia.

Effects of light finger touch on the regularity of center-of-pressure fluctuations during quiet bipedal and single-leg postural tasks.

BACKGROUND: The use of extra sources of sensory information associated with light fingertip touch to enhance postural steadiness has been associated with increased attentional demands, whereas the regularity of center of pressure (COP) fluctuations has been interpreted as a marker of the amount of attention invested in posture control. RESEARCH QUESTION: This study addressed whether increased attentional demands associated with postural tasks involving light finger touch might be reflected by measures of COP regularity. METHODS: The experiments involved quiet bipedal stance (n = 8 participants) and single-legged stance (n = 14 participants). Each participant was instructed to stand as quietly as possible on a force plate, either touching an external rigid surface (applied force < 1 N, light touch condition), or not (no touch condition). Postural steadiness was assessed by traditional COP measurements (COP Area, RMS, and velocity), whereas the regularity of postural sway was based on estimates of the sample entropy (SaEn) of the COP time series. RESULTS: Traditional parameters of postural sway and COP regularity (inversely related to SaEn COP measurements) were reduced during the touch conditions as compared to the no-touch conditions, for both bipedal quiet stance and single-legged stance. Decreased COP regularity with light touch was mainly reflected in the direction of the largest postural sway (i.e. in the sagittal plane for bipedal stance and in the frontal plane for single-legged stance). SIGNIFICANCE: The present results suggest that actively touching an external surface with the fingertip, besides increasing postural steadiness, generated an externally oriented (presumably cognitive-dependent) focus of attention, so that participants invested less attention on the postural task per se (as suggested by increased SaEn), which might be associated with a more "automatic" control of posture.

High levels of extracellular ATP lead to different inflammatory responses in COVID-19 patients according to the severity.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has significantly impacted the world and has driven many researchers into the pathophysiology of COVID-19. In the findings, there is a close association between purinergic signaling and the immune response. Then, this study aimed to evaluate alterations in the purinergic signaling in COVID-19 patients according to range severity. We divided the COVID-19 patients into moderate and severe cases following the guideless of NIH and WHO, together with clinical characteristics. The blood samples were collected to obtain PBMCs and platelets. We analyzed the ectonucleotidase activities through ATP, ADP, AMP, Ado hydrolysis, E-NTPDase1 (CD39), and 5'-NT (CD73) expression by flow cytometry in total leukocytes. The extracellular ATP was measured by bioluminescence, and cytokines were analyzed by flow cytometry. We observed a decrease in ATP hydrolysis and increased AMP hydrolysis in PBMCs for both groups. In severe cases, ATP hydrolysis was raised for the platelets, while ADP and AMP hydrolysis have risen significantly in both groups. Additionally, there was a significant increase in ADP hydrolysis in severe cases compared to moderate cases. In addition, we observed an increase in the ADA activity in platelets of moderate patients. Moderate and severe cases showed increased expression of CD39 and CD73 in total leukocytes. To finalize the purinergic signaling, extracellular ATP was increased in both groups. Furthermore, there was an increase in IL-2, IL-6, IL-10, and IL-17 in moderate and severe groups. Thus, for the first time, our findings confirm the changes in purinergic signaling and immune response in COVID-19, in addition to making it more evident that the severity range directly impacts these changes. Therefore, the therapeutic potential of the purinergic system must be highlighted and studied as a possible target for the treatment of SARS-CoV-2 disease. KEY MESSAGES: COVID-19 patients exhibit alterations in purinergic system and immune response. High levels of extracellular ATP lead to different inflammatory responses. CD39 and CD73 expression were increased in COVID-19 patients. Cytokines IL-2, IL-6, IL-10, and IL-17 also were altered in these patients. The purinergic system may be a possibility target to SARS-CoV-2 treatments.

Effects of Fatigue on Postural Sway and Electromyography Modulation in Young Expert Acrobatic Gymnasts and Healthy Non-trained Controls During Unipedal Stance.

This study investigated whether expert acrobatic gymnasts respond differentially than their non-trained counterparts during a single-legged stance task performed before and after a protocol designed to induce fatigue in the ankle plantarflexor muscles in terms of (a) postural steadiness and (b) electromyography (EMG) activation. We hypothesized that neuromuscular adaptation due to training would lead to different behavior of center of pressure (COP) and EMG quantifiers after fatigue. Twenty eight female volunteers (aged 11 to 24 years) formed two groups: expert acrobatic gymnastics athletes (GYN, n = 14) and age-matched non-gymnasts [control (CTRL), n = 14]. Fatigue of the ankle plantarflexors (dominant leg) was induced by a sustained posture (standing on the toes) until exhaustion. Traditional COP parameters (area, RMS, mean velocity, and power spectrum at low and high frequency ranges) were obtained with a force plate, and time and frequency-domain EMG parameters were obtained by surface electrodes positioned on the tibialis anterior, soleus, lateral gastrocnemius, medial gastrocnemius, vastus lateralis, biceps femoris, spinal erector and rectus abdominis muscles. The main results showed that fatigue induced a significant increase in postural oscillations in the ML axis (including RMS, velocity and frequency components of the power spectrum), with no significant effects in the AP axis. In terms of postural sway parameters (i.e., COP quantifiers), no superior balance stability was found for the GYN group as compared to CTRL, irrespective of the fatigue condition. On the other hand, the modulation of EMG parameters (in both time and frequency domains) indicated that expert acrobatic gymnastics athletes (as compared to healthy untrained matched controls) used different neuromuscular control strategies to keep their postures on single-legged quiet standing after the fatiguing protocol. The present results improve our knowledge of the mechanisms behind the interplay between fatigue and postural performance associated with the neuromuscular adaptations induced by sport practice. The design of gymnastics training might consider strategies aimed at improving the performance of specific muscles (i.e., tibialis anterior, soleus, biceps femoris, spinal erector) for which particular activation patterns were used by the acrobatic gymnastics to control single-legged quiet standing.

In vitro and in silico protein corona formation evaluation of curcumin and capsaicin loaded-solid lipid nanoparticles.

Nanotechnology has been an important tool for the production of nanoparticles with controlled release of drugs for therapeutic applications. Here, we produced solid lipid nanoparticles (SLN) loaded with curcumin and capsaicin (NCC) following the overarching goals of green chemistry. Currently, besides evaluating the composition, and size of these, it is necessary to understand the interactions between nanoparticles and the biomolecules present in the biological medium. For this, assays were conducted in order to evaluate the potential formation of the protein 'corona', and to better understand the results obtained in vitro, we also performed an interaction study, in silico, between the NCC components and the main serum protein, albumin. In the first hour of contact between the NCC and the culture medium showed fluctuation in the diameter of the NCC. However, after 24 and 48 h of the incubation period, all NCC concentrations showed an increase in size, which can be attributed to plasma protein adsorption. Since, hard corona takes a few seconds, while the soft corona can be formed in minutes up to a few hours. On the other hand, best docking binding-poses of interaction for the formed docking complexes evaluated suggest interactions following the docking affinity like free energy FEB (Tween 80-bovine serum albumin) ≈ FEB (Span 80-bovine serum albumin) showing a pharmacodynamic pattern based in non-covalent hydrophobic interactions with the bovine serum albumin binding-site. Our in silico results clarify and reinforce our in vitro findings of corona formation, which represents the real interaction with cell membranes in vivo.

A case report on catamenial epilepsy / Philippine Journal of Obstetrics and Gynecology

A case of a 17-year-old nulligravid with onset of seizure episodes since menarcheis reported. She was diagnosed with Seizure Disorder treated with Phenobarbital and was seizure free for 2 years. Two years prior to consult, seizure recurrences were noted to coincide with menstruation, hence, was diagnosed with Catamenial Epilepsy. Patient was shifted to Lamotrigine but seizure exacerbations were still observed, prompting referral to the Reproductive Medicine service for adjunctive hormonal therapy. Depot medroxyprogesterone acetate was added to the antiepileptic drug which provided seizure control. Adjunctive hormonal therapy proved to be helpful in the management of intractable seizures in this patient. The report aims to give a better understanding of the neuroactive properties of estrogen and progesterone and its role in the development of Catamenial Epilepsy. Gender-related and psychosocial issues in the treatment of Epilepsy in the child-bearing years up to the menopause are also discussed.

A case report on catamenial epilepsy / Philippine Journal of Obstetrics and Gynecology

A case of a 17-year-old nulligravid with onset of seizure episodes since menarcheis reported. She was diagnosed with Seizure Disorder treated with Phenobarbital and was seizure free for 2 years. Two years prior to consult, seizure recurrences were noted to coincide with menstruation, hence, was diagnosed with Catamenial Epilepsy. Patient was shifted to Lamotrigine but seizure exacerbations were still observed, prompting referral to the Reproductive Medicine service for adjunctive hormonal therapy. Depot medroxyprogesterone acetate was added to the antiepileptic drug which provided seizure control. Adjunctive hormonal therapy proved to be helpful in the management of intractable seizures in this patient.The report aims to give a better understanding of the neuroactive properties of estrogen and progesterone and its role in the development of Catamenial Epilepsy. Gender-related and psychosocial issues in the treatment of Epilepsy in the child-bearing years up to the menopause are also discussed.

Profiles of Entamoeba histolytica-specific immunoglobulins in human sera.

OBJECTIVE: To determine the profiles of anti-Entamoeba histolytica (E. histolytica) IgA, IgG, and IgM in sera of diarrheic and non-diarrheic individuals and partially characterize target antigens. METHODS: Serum samples from thirty diarrheic and thirty non-diarrheic individuals were subjected to IgA, IgG, and IgM profiling through enzyme-linked immunosorbent assay (ELISA), flow cytometry, and immunoblot. RESULTS: ELISA titer results showed that both diarrheic and non-diarrheic individuals possess high levels of E. histolytica-specific IgG compared to IgA and IgM. Flow cytometry data showed that diarrheic serum samples had higher mean reaction percentages against E. histolytica cells compared to non-diarrheic samples. Immunoreactive E. histolytica proteins with molecular weights ranging between 7 kDa and 292 kDa were recognized by diarrheic serum IgG, and 170 kDa and 250 kDa by non-diarrheic serum IgG. CONCLUSIONS: Our findings suggest that serum anti-E. histolytica IgG, compared with serum anti-E. histolytica IgA and IgM responses, was generally high in both diarrheic and non-diarrheic sera, indicating a past exposure to the organism both in symptomatic patients as well as in asymptomatic carriers, respectively. In addition, serum IgG from diarrheic and non-diarrheic patients were able to detect immunogenic E. histolytica proteins.

Auditory and oral language abilities in children with cochlear implants: a case study.

The cochlear implant (CI) represents the most important advance in the treatment of individuals with severe to profound bilateral hearing loss who do not benefit from hearing aids. Children who receive the CI during the critical period of neuroplasticity of the auditory system, when combined with speech therapy, have the chance to develop the auditory and linguistic skills similarly to their normal hearing peers. Two cases of implanted children are presented in this study, and one of them was not enrolled in a formal aurioral therapeutic program since the implantation surgery. At the moment of language and auditory assessment, the children were 2 years and 5 months old, and the CI had been used for 11 months. According to the results presented, it was observed that the child enrolled in rehabilitation program had better auditory and language performance when compared to the other child. Despite the remarkable benefits that the CI provides to children with hearing impairment, the device itself only provides the child with the audibility of environmental sounds and speech signal. For the auditory and language development to happen, it is necessary, among other factors, a speech-language intervention, with partnership between professionals and parents.

Profiles of Entamoeba histolytica-specific immunoglobulins in human sera

OBJECTIVE@#To determine the profiles of anti-Entamoeba histolytica (E. histolytica) IgA, IgG, and IgM in sera of diarrheic and non-diarrheic individuals and partially characterize target antigens.@*METHODS@#Serum samples from thirty diarrheic and thirty non-diarrheic individuals were subjected to IgA, IgG, and IgM profiling through enzyme-linked immunosorbent assay (ELISA), flow cytometry, and immunoblot.@*RESULTS@#ELISA titer results showed that both diarrheic and non-diarrheic individuals possess high levels of E. histolytica-specific IgG compared to IgA and IgM. Flow cytometry data showed that diarrheic serum samples had higher mean reaction percentages against E. histolytica cells compared to non-diarrheic samples. Immunoreactive E. histolytica proteins with molecular weights ranging between 7 kDa and 292 kDa were recognized by diarrheic serum IgG, and 170 kDa and 250 kDa by non-diarrheic serum IgG.@*CONCLUSIONS@#Our findings suggest that serum anti-E. histolytica IgG, compared with serum anti-E. histolytica IgA and IgM responses, was generally high in both diarrheic and non-diarrheic sera, indicating a past exposure to the organism both in symptomatic patients as well as in asymptomatic carriers, respectively. In addition, serum IgG from diarrheic and non-diarrheic patients were able to detect immunogenic E. histolytica proteins.

Habilidades auditivas e linguísticas iniciais em crianças usuárias de implante coclear: relato de caso / Auditory and oral language abilities in children with cochlear implants: a case study

O implante coclear (IC) representa o mais importante avanço no tratamento de deficientes auditivos de grau severo e/ou profundo bilateral que não apresentam aproveitamento com o aparelho de amplificação sonora individual. A realização do IC no período crítico da neuroplasticidade do sistema auditivo possibilita que o desenvolvimento auditivo e linguístico de crianças implantadas precocemente sejam semelhantes ao desenvolvimento destas habilidades em crianças ouvintes, quando associadas à terapia fonoaudiológica. Os casos a serem apresentados referem-se a duas crianças usuárias de IC, sendo que uma delas não estava inserida em programa terapêutico com abordagem aurioral desde a realização da cirurgia do dispositivo. No momento da aplicação dos protocolos para avaliação das habilidades auditivas e de linguagem, as crianças estavam com 2 anos e 5 meses de idade e 11 meses de uso do IC. De acordo com os resultados apresentados pelos casos estudados foi possível observar que a criança inserida em programa terapêutico apresentou melhor desempenho auditivo e linguístico, comparada ao desenvolvimento da outra criança. Apesar dos notáveis benefícios que o IC proporciona nas crianças deficientes auditivas, o dispositivo por si só proporciona à criança audibilidade aos sons ambientais e de fala. Para que o desenvolvimento auditivo e linguístico aconteça torna-se necessário, dentre outros fatores, a realização da terapia fonoaudiológica, com parceria entre profissionais e pais.

The cochlear implant (CI) represents the most important advance in the treatment of individuals with severe to profound bilateral hearing loss who do not benefit from hearing aids. Children who receive the CI during the critical period of neuroplasticity of the auditory system, when combined with speech therapy, have the chance to develop the auditory and linguistic skills similarly to their normal hearing peers. Two cases of implanted children are presented in this study, and one of them was not enrolled in a formal aurioral therapeutic program since the implantation surgery. At the moment of language and auditory assessment, the children were 2 years and 5 months old, and the CI had been used for 11 months. According to the results presented, it was observed that the child enrolled in rehabilitation program had better auditory and language performance when compared to the other child. Despite the remarkable benefits that the CI provides to children with hearing impairment, the device itself only provides the child with the audibility of environmental sounds and speech signal. For the auditory and language development to happen, it is necessary, among other factors, a speech-language intervention, with partnership between professionals and parents.