Low-Level Expression of p-S6 Is Associated with Nodal Metastasis in Patients with Head and Neck Cutaneous Squamous Cell Carcinoma.

Cutaneous squamous cell carcinoma (cSCC) is the second most common form of skin cancer. The incidence of metastasis for cSCC is estimated to be around 1.2-5%. Ribosomal protein S6 (p-S6) and the p21 protein (p21) are two proteins that play central roles in other cancers. These proteins may be equally important in cSCC, and together, these could constitute a good candidate for metastasis risk assessment of these patients. We investigate the relationship of p-S6 and p21 expression with the impact on the prognosis of head and neck cSCC (cSCCHN). p-S6 and p21 expression was analyzed by immunohistochemistry on paraffin-embedded tissue samples from 116 patients with cSCCHN and associations sought with clinical characteristics. Kaplan-Meier estimators and Cox proportional hazard regression models were also used. The expression of p-S6 was significantly inversely associated with tumor thickness, tumor size, desmoplastic growth, pathological stage, perineural invasion and tumor buds. p21 expression was significantly inversely correlated with >6 mm tumor thickness, desmoplastic growth, and perineural invasion. p-S6-negative expression significantly predicted an increased risk of nodal metastasis (HR = 2.63, 95% CI 1.51-4.54; p < 0.001). p21 expression was not found to be a significant risk factor for nodal metastasis. These findings demonstrate that p-S6-negative expression is an independent predictor of nodal metastasis. The immunohistochemical expression of p-S6 might aid in better risk stratification and management of patients with cSCCHN.

Controlling solid-liquid-solid GeSn nanowire growth modes by changing deposition sequences of a-Ge:H layer and SnO2nanoparticles.

Alloying Ge with Sn is one of the promising ways for achieving Si compatible optoelectronics. Here, GeSn nanowires (NWs) are realized via nano-crystallization of a hydrogenated amorphous Ge (a-Ge:H) layer with the help of metal Sn droplets. The full process consists of three steps: (1) SnO2nanoparticle (NP) reduction in a hydrogen plasma to produce Sn catalyst; (2) a-Ge:H deposition at 120 °C and (3) annealing. GeSn alloys with rich morphologies such as discrete nanocrystals (NCs), random, and straight NWs were successfully synthesized by changing process conditions. We show that annealing under Ar plasma favors the elaboration of straight GeSn NWs in contrast to the conventional random GeSn NWs obtained when annealing is performed under a H2atmosphere. Interestingly, GeSn in the form of discrete NCs can be fabricated during the deposition of a-Ge:H at 180 °C. Even more, the synthesis of out-of-plane GeSn NWs has been demonstrated by reversing the deposition sequence of SnO2NPs and a-Ge:H layer.

Ustekinumab Drug Survival in Patients with Psoriasis: A retrospective Study of Real Clinical Practice.

Background and objectives: The efficacy and safety of ustekinumab have been proved in clinical trials. In daily clinical practice, knowing the factors that determine survival differences of biological drugs allows psoriasis treatment to be optimized as a function of patient characteristics. The main objectives of this work are to understand ustekinumab drug survival in patients diagnosed with plaque psoriasis in the Hospital Universitario Central de Asturias (HUCA Dermatology Department, and to identify the predictors of drug discontinuation. Materials and Methods: A retrospective hospital-based study, including data from 148 patients who were receiving ustekinumab (Stelara®) between 1 February 2009 and 30 November 2019, were collected. Survival curves were approximated through the Kaplan-Meier estimator and compared using the log-rank test. Proportional hazard Cox regression models were used for multivariate analyses while both unadjusted and adjusted hazard ratios (HR) were used for summarizing the studied differences. Results: The average duration of the treatment before discontinuation was 47.57 months (SD 32.63 months; median 41 months). The retention rates were 82% (2 years), 66% (5 years), and 58% (8 years). Median survival was 80 months (95% confidence interval. CI 36.9 to 123.01 months). The survival study revealed statistically significant differences between patients with arthritis (log-rank test, p < 0.001) and those who had previously received biological treatment (log-rank test, p = 0.026). The five-year prevalence in patients still under treatment was 80% (those without arthritis) and 54% (arthritis patients). In the multivariate analysis, only the patients with arthritis had a lower rate of drug survival. No statistically significant differences were observed for any of the other comorbidities studied. The first and second most frequent causes of discontinuation were secondary failure and arthritis inefficacy, respectively. Conclusion: Ustekinumab is a biological drug conferring high survival in plaque psoriasis patients. Ustekinumab survival is lower in patients with arthritis.

Impact of neonatal malnutrition on expression TLR-9, NF-kB and cytokines of macrophages infected in vitro with methicillin resistant Staphylococcus aureus.

Early nutritional aggressions promote epigenetic adjustments that culminate in the loss of phenotype plasticity (with permanent long-term modifications). Maternal diet and inadequate neonatal nutrition can result in fetal programming that presents susceptibility to infections in adult life. Thus, it becomes essential to verify the impacts of neonatal malnutrition (even following nutritional replacement) on the immunological response to methicillin resistant Staphylococcus aureus (MRSA) infections. Male rats were divided into two distinct groups: Nourished and Malnourished. After isolation of mononuclear cells, four systems were established: negative control, positive control and two testing systems, (MSSA and MRSA). Tests were performed to analyze expression of TLR-9, NF-kB, IL-1ß, IL-18 and IL-33. For statistical analysis, we used the Student t and ANOVA tests p < 0.05. Even after nutritional replacement, malnutrition in the neonatal period compromised the animals' weight gains p < 0.05. There was a reduction in the expression of the immunological response in the positive control, however deregulation was observed in the gene expression of MRSA-infected macrophages, with a reduction in TLR-9 expression, and overexpression in NF-kB and cytokines p < 0.05. Puppies inflicted with protein-calorie malnutrition were compromised; (long-term) body growth and immune response. In the infectious scenario, immune collapse is reflected in inflammatory response exacerbation with a likely histolytic character. Immune disabling (resulting from gene expression deregulation) causes susceptibility to infections due to ineffective recognition, intense pro-inflammatory mediation, and cell death. It is suggested that neonatal malnutrition can program susceptibility to multiresistant bacterial infections, and generally favors a triggering of more intense confrontations with fatal outcomes.

Morphofunctional Effect of Stem Cells on the Regeneration of the Facial Nerve in a Rat Model.

PURPOSE: The aim of the present study was to determine the clinical, histologic, and histomorphometric influence of stem cells on the regeneration of facial nerves in rats submitted to neurotmesis with 5-mm defects. MATERIALS AND METHODS: Thirty-six male Wistar rats were submitted to neurotmesis. In group 1 (n = 18), the 2 extremities of the nerve trunk were sutured in a polyethylene tube containing a protein matrix (Matrigel; Invitrogen, BD Biosciences, Franklin Lakes, NJ). Group 2 (n = 18) underwent the same procedures with the incorporation of stem cells in the tube. The groups were subdivided by the day of euthanasia (30, 60, and 90 days). Evaluations of the central portion and distal extremity of the tubes were performed in both groups. RESULTS: During the functional evaluation, group 2 exhibited better closing of the eyelids at 30 and 60 days. The histomorphometric analyses revealed a significant reduction in the area of the nerve fibers in group 1 compared with group 2 at all 3 evaluation (P = .031). In group 2, the area of the nerve fibers was significantly greater (P = .031), and the thickness of the myelin sheath was significantly greater in the center (P = .002) and distal (P = .019) extremity of the tube at 30 days compared with the findings in group 1. CONCLUSIONS: The present findings suggest that the use of stem cells in nerve injuries will result in faster, more effective regeneration within the intervals analyzed.

Functionality and quality of life of patients with unilateral lymphedema of a lower limb: a cross-sectional study.

BACKGROUND: Lymphedema of the lower limbs is a chronic disease caused by damage to the lymphatic system that influences people's mobility, functionality, and quality of life. Questionnaires and physical test are very practical, easy to apply, and low cost methods that provide important data for evaluation of these patients. OBJECTIVES: To evaluate the influence of unilateral lower limb lymphedema on functionality and quality of life, correlating 3 assessment tools. METHODS: This was a descriptive study investigating 25 patients of both sexes with unilateral lymphedema in a lower limb. Limb volume was assessed using circumferential tape measurements, the Medical Outcomes Study Short Form-36 Health Survey (SF-36) was used to assess quality of life, the Lymphoedema Functioning, Disability and Health Questionnaire for Lower Limb Lymphoedema (Lymph-ICF-LL) was used to assess physical, mental, and social skills related to lymphedema, and the Timed Up and Go (TUG) test was used for functional assessment. RESULTS: Lymphedema was present throughout the affected lower limb of participants. The domains most affected by lymphedema were physical aspects (25.0 ± 31.4) and emotional aspects (36.0 ± 42.9) from the SF-36 and the mobility domain (6.0 ± 2.6) from the Lymph -ICF-LL. Patients performed the TUG in 9.88 ± 1.98 seconds. The TUG was correlated with the questionnaires and the questionnaires were correlated with each other. CONCLUSIONS: People with unilateral lower limb lymphedema exhibited negative impacts on quality of life and functionality, as evaluated by questionnaires, which were correlated with each other. TUG performance was within normal limits, but results correlated with the questionnaires used.

All-trans Retinoic Acid Augments Autophagy during Intracellular Bacterial Infection.

Vitamin A deficiency strongly predicts the risk of developing tuberculosis (TB) in individuals exposed to Mycobacterium tuberculosis (Mtb). The burden of antibiotic-resistant TB is increasing globally; therefore, there is an urgent need to develop host-directed adjunctive therapies to treat TB. Alveolar macrophages, the niche cell for Mtb, metabolize vitamin A to all-trans retinoic acid (atRA), which influences host immune responses. We sought to determine the mechanistic effects of atRA on the host immune response to intracellular bacterial infection in primary human and murine macrophages. In this study, atRA promoted autophagy resulting in a reduced bacterial burden in human macrophages infected with Mtb and Bordetella pertussis, but not bacillus Calmette-Guérin (BCG). Autophagy is induced by cytosolic sensing of double-stranded DNA via the STING/TBK1/IRF3 axis; however, BCG is known to evade cytosolic DNA sensors. atRA enhanced colocalization of Mtb, but not BCG, with autophagic vesicles and acidified lysosomes. This enhancement was inhibited by blocking TBK1. Our data indicate that atRA augments the autophagy of intracellular bacteria that trigger cytosolic DNA-sensing pathways but does not affect bacteria that evade these sensors. The finding that BCG evades the beneficial effects of atRA has implications for vaccine design and global health nutritional supplementation strategies. The ability of atRA to promote autophagy and aid bacterial clearance of Mtb and B. pertussis highlights a potential role for atRA as a host-directed adjunctive therapy.

Nanoscale control of Si nanoparticles within a 2D hexagonal array embedded in SiO2 thin films.

In this work, we investigate the ability to control Si nanoparticles (NPs) spatially arranged in a hexagonal network of 20 nm wide nanovolumes at controlled depth within SiO2 thin films. To achieve this goal an unconventional lithographic technique was implemented based on a bottom-up approach, that is fully compatible with the existing semiconductor technology. The method combines ultra-low energy ion beam synthesis with nanostructured block-copolymer thin films that are self-assembled on the SiO2 substrates to form a nanoporous template with hexagonally packed pores. A systematic analytical investigation using time of flight-secondary ion mass spectroscopy and low-loss energy filtered transmission electron microscopy demonstrates that by adjusting few fabrication parameters, it is possible to narrow the size distribution of the NPs and to control the number of NPs per nanovolume. Experimental results are critically discussed on the basis of literature data, providing a description of the mechanism involved in the formation of Si NPs.

Relation between neonatal malnutrition and gene expression: inflammasome function in infections caused by Candida Albicans.

PURPOSE: To investigate the effects of neonatal malnutrition followed by nutritional replacement on the signaling mechanisms developed by the inflammasome complex by analyzing the expression of the targeted TLR2, TLR4, NLRP3, caspase-1 and release of IL-1ß and IL-18 by alveolar macrophages infected in vitro with Candida albicans. METHODS: Male Wistar rats (n = 24), 90-120 days, were suckled by mothers whose diet during lactation contained 17 % protein in the nourish group and 8 % protein in the malnourished group. After weaning, both groups were fed a normal protein diet. Macrophages were obtained after tracheostomy, through the collection of bronchoalveolar lavage fluid. The quantification of the expression levels of targets (TLR2, TLR4, NLRP3 and caspase-1) was performed by real-time RT-PCR. Production of cytokines was performed by ELISA. RESULTS: The malnourished animals during lactation showed reduced body weight from the fifth day of life, remaining until adulthood. Further, the model applied malnutrition induced a lower expression of TLR4 and caspase-1. The quantification of the TLR2 and NLRP3, as well as the release of IL-1ß and IL-18, was not different between groups of animals nourished and malnourished. The system challenged with Candida albicans showed high expression levels of all targets in the study. CONCLUSIONS: The tests demonstrate nutritional restriction during critical periods of development, although nutritional supplementation may compromise defense patterns in adulthood in a timely manner, preserving distinct signaling mechanism, so that the individual does not become widely vulnerable to infections by opportunistic pathogens.

Neonatal malnutrition programs the oxidant function of macrophages in response to Candida albicans.

Experimental maternal nutrition restriction models are used to investigate short or long-term consequences of nutritional deficiency on puppies' growth. By assuming that the immune function is directly related to host's nutritional status, the current study aims to investigate the effects of neonatal malnutrition on oxidative stress and on the cell death of the alveolar macrophage after in vitro infection by Candida albicans. Wistar rats were suckled by mothers fed on diets containing 17% protein (Nourished group) or 8% protein (Malnourished group) in the current assay. Both groups received the standard diet used in the vivarium until adulthood, after weaning. The results showed that the offspring from mothers fed on low-protein diet presented lower body weight from 5 days of life on. Their low weight remained until adulthood when it was compared to that of rats in the nourished group. Superoxide and nitric oxide production was lower in malnourished animals and it was accompanied by low inducible nitric oxide synthase gene expression levels in systems in which the alveolar macrophages were challenged by immunogenic stimulus. No significant differences were observed in comparisons performed between the nourished and malnourished groups in any of the analyzed cell viability (apoptosis/necrosis) parameters. The fungal inoculum-stimulated system induced higher oxidative stress and cell death by necrosis. The current study demonstrated that dietary restriction during lactation alters the oxidant function of alveolar macrophages in puppies; It happens from the gene transcription step to the release of mediators, thus compromising the host's defenses against Candida albicans. It raises the possibility that Candida albicans may cease to be a commensal fungus to become a pathogen in offspring that have suffered nutritional deficiency during critical developmental periods, due to impaired immune responses.

Effect of neonatal malnutrition on expression of nitric oxide synthase enzyme, production of free radicals and in vitro viability of alveolar macrophages infected with methicillin-sensitive and methicillin-resistant Staphylococcus aureus.

OBJECTIVE: Evaluate the effects of neonatal malnutrition on the microbicidal response and viability of in vitro macrophages infected with Staphylococcus aureus sensitive/resistant to methicillin. METHODS: Male Wistar rats (n = 24) were divided into two distinct groups: nourished (rats breast-fed by mothers undergoing diet with 17% casein) and malnourished (rats breast-fed by mothers undergoing diet with 8% casein). Macrophages were recovered after surgical tracheostomy procedure by collecting bronchoalveolar lavage. Four systems were established: negative control, composed only by phagocytes; positive control, macrophages plus lipopolysaccharide; and two test systems, macrophages plus Staphylococcus aureus sensitive and resistant to methicillin. Plates were incubated at 37 °C for 24 h. After this period, tests for the analysis of cell viability and microbicidal response were performed. In the statistical analysis, the Student's t and ANOVA tests were used, accepting p < 0.05. RESULTS: The neonatal malnutrition impaired the animals' body weight. There was a lower expression of the inducible nitric oxide enzyme (iNOS), nitric oxide production, and viability of macrophages infected with methicillin-resistant Staphylococcus aureus. However, increased production of superoxide anion in the malnourished group was detected. CONCLUSION: Neonatal malnutrition focusing on critical periods of development promoted lower expression of iNOS, nitric oxide production, cell viability, and exacerbated reactive oxygen species production. The high levels of reactive oxygen species may favor the onset of serious and systemic infections with fatal outcome if associated with methicillin-resistant Staphylococcus aureus.

Biocompatibility and cutaneous reactivity of cellulosic polysaccharide film in induced skin wounds in rats.

The development of a skin substitute suitable for immediately performing the function of the lost dermis and epidermis could result in a positive impact on the treatment of patients with extensive skin lesions. A biopolymer film was applied to skin wounds to investigate the biocompatibility and cutaneous reaction and to test its activity as a mechanical barrier and conductor in the healing process. Forty Wistar rats of both sexes were used in the present study. Two excisions were performed in the dorsal part of the skin flaps. The polysaccharide film was applied over one of the incisions and other incision was washed with saline. The time spent for complete healing of both lesions was virtually the same in both groups, during 21 days of observation. The film remained attached to the bed of the exposed wound for an average period of 6 days. There were no statistically significant differences with regard to lesion measurement area at assessment times of 2nd, 7th and 14th postoperative days. At day 21, the scar area showed a significant difference (0.0229). After 40 days, all wounds were completely healed. No statistically significant differences were found between the histological parameters assessed in the experimental and control groups. The cellulosic polysaccharide film integrated well with the tissue showing high biocompatibility and low skin reactivity.

Expectorant and antioxidant activities of purified fumarprotocetraric acid from Cladonia verticillaris lichen in mice.

UNLABELLED: The lichen Cladonia verticillaris produces bioactive secondary metabolites, such as fumarprotocetraric (FUM) and protocetraric acids. Species of the genus Cladonia demonstrate anti-tumor, anti-inflammatory and antipyretic activities and have been used in folk medicine to treat respiratory diseases (throat irritation, cough, asthma and tuberculosis). The aim of the present study was to evaluate the expectorant and mucolytic activities of fumarprotocetraric acid in albino Swiss mice. FUM was extracted and purified from an acetone extract of C. verticillaris. The phenol red quantification method was used on the bronchoalveolar lavage fluid following the administration of FUM (25, 50 or 100 mg/kg orally or intraduodenally and 12.5, 25 or 50 mg/kg, intraperitoneally) for the evaluation of expectorant activity. Control groups received either saline solution (7.5 mL/kg) or ambroxol (1 mg/kg) through the same administration routes. Antioxidant activity was evaluated using the thiobarbituric acid reactive species assay in mouse lung tissue treated with the FUM at 25, 50 or 100 mg/kg orally, followed by a lipopolysaccharide solution at 1 mg/kg intrapleurally. The same protocol was used for the control groups using either saline solution (7.5 mL/kg, orally) or N-acetylcysteine (20 mg/kg, orally). RESULTS: Orally administered FUM at doses of 25 and 50 mg/kg promoted significantly greater dose-dependent phenol red activity in the bronchoalveolar lavage and expectorant activity in comparison to the controls (p < 0.05). Lipid peroxidation (malondialdehyde equivalent) was reduced by 50% in the lung tissue. CONCLUSION: The results confirm the expectorant and antioxidant properties of fumarprotocetraric acid produced by the lichen C. verticillaris.

Autophagy regulates IL-23 secretion and innate T cell responses through effects on IL-1 secretion.

Autophagy controls IL-1ß secretion by regulating inflammasome activation and by targeting pro-IL-1ß for degradation. In this article, we show that inhibition of autophagy, either with the PI3K inhibitors 3-methyladenine, wortmannin, and LY294002 or with small interfering RNA against autophagy proteins augmented the secretion of IL-23 by human and mouse macrophages and dendritic cells in response to specific TLR agonists. This process occurred at the transcriptional level and was dependent on reactive oxygen species and IL-1R signaling; it was abrogated with an IL-1R antagonist or with IL-1-neutralizing Abs, whereas treatment with either rIL-1α or IL-1ß induced IL-23 secretion. Dendritic cells treated with LPS and 3-methyladenine secreted enhanced levels of both IL-1ß and IL-23, and supernatants from these cells stimulated the innate secretion of IL-17, IFN-γ, and IL-22 by γδ T cells. These data demonstrate that autophagy has a potentially pivotal role to play in the induction and regulation of inflammatory responses by innate immune cells, largely driven by IL-1 and its consequential effects on IL-23 secretion.

Intratracheal co-administration of antioxidants and ceftriaxone reduces pulmonary injury and mortality rate in an experimental model of sepsis.

BACKGROUND AND OBJECTIVE: Recent studies showed that both sepsis and antibiotic therapy are associated with cell death and linked to reactive oxygen species generation. This study investigated the effects of intratracheal administration of combinations of antioxidants (n-acetyl cysteine (NAC), vitamins C and E) in the treatment of sepsis-induced lung injury. METHODS: Ninety-six male Wistar rats subjected to sepsis were treated with ceftriaxone plus NAC with or without vitamins C and E and compared to appropriate controls. As an index of oxidative damage protein carbonyls, sulfhydryl groups, lipid peroxidation and superoxide anion were measured, as well as superoxide dismutase and catalase. Histopathological alterations and mortality rate were also analyzed. RESULTS: Twenty-four hours after sepsis induction, markers of oxidative stress increased in all lungs examined. Ceftriaxone plus intratracheal combination of NAC, vitamins C and E decreased lung injury in infected animals by reducing superoxide anion production (54%), lipid peroxidation (53%) and protein carbonyl (58%) and restored the redox status (7.5 times). This therapy also reduced the imbalance of antioxidant enzymes activities and attenuated the alveolar architectural disorganization, inflammatory cell infiltration and pulmonary oedema. Survival increased from 66.6% with ceftriaxone to 83.2% with ceftriaxone plus antioxidants. CONCLUSIONS: Ceftriaxone plus intratracheal co-administration of antioxidants provides better protection, by decreasing pulmonary oxidative stress, limiting histophatological alterations and improving survival. Antioxidants should be explored as a co-adjuvant in the treatment of severe lung injury.

Effect of surgical repair of the joint capsule in mandibular condyle fractures in adult rats.

PURPOSE: The aim of the present study was to assess histologic changes in the temporomandibular joint (TMJ) of adult rats subjected to unilateral fracture of the mandibular condyle and soft tissue injury. MATERIALS AND METHODS: The animals were divided into 2 groups: one had surgical treatment for soft tissue repair and the other had no soft tissue treatment. All histologic evaluations were performed according to the presence or absence of synovitis, vascularity, presence or absence of joint inflammation, and presence or absence of the articular disc. The contralateral TMJs also were evaluated. RESULTS: The results showed few histologic changes in the synovial membrane and joint disc for the 2 groups and in the synovial membrane and disc of the contralateral side, where indirect trauma occurred in the unoperated joint. CONCLUSION: This study showed that treating or not treating soft tissues does not change the treatment results of condyle fracture or interfere with TMJ pathosis.