Testicular subcutaneous allografting followed by immunosuppressive treatment promotes maintenance of spermatogonial cells in rainbow trout (Oncorhynchus mykiss).

Germ cell transplantation and testis graft represent promising biotechnologies that can be applied for the reproduction of commercial or endangered species. However, mechanisms of rejection from the host immune system might remove the transplanted donor cells/tissues and limit the surrogate production of gametes. In this work, we administered emulsion containing-immunosuppressants to verify whether they are capable to prevent immune rejection and promote survival of testis allografts in rainbow trout. In the first part of this study, we demonstrated in vitro that tacrolimus and cyclosporine were able to affect viability, inhibit leucocyte proliferation, and suppress il2 expression in vitro. In in vivo experiments, both doses of tacrolimus (0.5 and 1.5 mg/kg) and the lower dose of cyclosporine (20 mg/kg) significantly inhibited the expression of il2 in head kidney, three days post-injection. A higher dose of cyclosporine (40 mg/kg) was able to inhibit il2 expression for up to seven days post-injection. In the second part, testis allografts were conducted in fish treated weekly with emulsion containing-tacrolimus. Immunohistochemical, conventional histology, and qRT-PCR (vasa) analysis demonstrated the presence of spermatogonial cells by the fifth week, in animals treated with 0.5 mg/kg of tacrolimus similar as found in autografted group. In the group treated with the highest tacrolimus dose (1.5 mg/kg) and in the non-treated group (without immunosuppressant), no germ cells or their respective markers were detected. il2 expression in head kidney was also suppressed in grafted animals treated with tacrolimus compared to non-treated group. These results suggest that tacrolimus may be a promising immunosuppressant for testis allografts or germ cell transplantation in rainbow trout. Co-administration combining tacrolimus (at lower dose) with other immunosuppressive drugs for inhibiting other activation pathways of the immune system, as performed in human organ transplantation, could be an alternative approach to optimize the immunosuppressive effects in host organisms.

Cinemeducation in clinical pharmacology: using cinema to help students learn about pharmacovigilance and adverse drug reactions.

PURPOSE: Feature films are increasingly being used in teaching health sciences. However, few publications address the effectiveness of this approach. We hypothesized that using feature films could help students learn. We aimed to assess the effectiveness of using a feature film to teach students about adverse drug reactions and pharmacovigilance. METHODS: The study population comprised third-, fifth-, and sixth-year undergraduate students of medicine, third-year undergraduate students of human biology, and graduate students in a master's degree program about the pharmaceutical and biotechnology industry. Students watched clips from the film 150 Miligrams (La fille de Brest) and discussed them afterward. To measure learning, we administered a 10-question multiple-choice test about pharmacovigilance concepts. We assessed students' satisfaction with the activity through a questionnaire. An exploratory comparative analysis was performed. RESULTS: A total of 237 students participated. Postintervention assessment scores were significantly higher than preintervention scores for the entire population and for all subgroups. The mean number of correct answers was 4.41 on the preintervention assessment and 5.78 on the postintervention assessment (mean gain: 1.37; 95% CI: 1.10-1.65). Similar results were found when analyzing groups of students from each group. Student satisfaction with this teaching activity was high in all groups. CONCLUSIONS: Cinemeducation is a useful tool for teaching about adverse drug reactions and pharmacovigilance processes. Most students were highly satisfied.

AC/DC Current Sensor for Rotating Applications.

There are several techniques for current measurement. Most of them are capable of measuring both alternating and direct current (AC/DC) components. However, they have severe drawbacks for rotating applications (large size, sensitivity to external fields, and low signal amplitude). In addition to these weaknesses, measured signals should be transmitted to a stationary part. In order to contribute solving these difficulties, this paper presents a sensor that can measure AC/DC simultaneously based on the electromagnetic coupling of two coils. To this aim, the measured waveform is analysed. In this paper, the design of such a sensor is presented. This design is validated through computer simulations and a prototype is built. The performance of this sensor prototype is analysed through experimental tests.

Performance Problems of Non-Toroidal Shaped Current Transformers.

Nowadays, non-toroidal shape primary pass-through current transformers are commonly used for large current machines with several cables per phase. As these transformers exhibit no radial symmetry, it is not clear if they can be tested using the indirect test described in the IEC 61869 standard. In order to answer this question, two non-toroidal shaped current transformers of different secondary winding designs have been tested and simulated. One transformer has a uniformly distributed secondary winding and the other has a partially distributed secondary winding. Both transformers have the same nameplate characteristics. Both perform correctly in the indirect test. However, only the transformer with the uniformly distributed secondary winding performs correctly in a direct test. A finite element simulation shows that the iron core of the partially distributed secondary winding transformer was saturated, while the iron core of the uniformly distributed one was not. This result explains their different performance. The main conclusion is that the indirect test is not sensitive enough to cover all cases and therefore under doubtful situations, the transformers should be tested using the direct test.

Connexins, Pannexins, and Their Channels in Fibroproliferative Diseases.

Cellular and molecular mechanisms of wound healing, tissue repair, and fibrogenesis are established in different organs and are essential for the maintenance of function and tissue integrity after cell injury. These mechanisms are also involved in a plethora of fibroproliferative diseases or organ-specific fibrotic disorders, all of which are associated with the excessive deposition of extracellular matrix components. Fibroblasts, which are key cells in tissue repair and fibrogenesis, rely on communicative cellular networks to ensure efficient control of these processes and to prevent abnormal accumulation of extracellular matrix into the tissue. Despite the significant impact on human health, and thus the epidemiologic relevance, there is still no effective treatment for most fibrosis-related diseases. This paper provides an overview of current concepts and mechanisms involved in the participation of cellular communication via connexin-based pores as well as pannexin-based channels in the processes of tissue repair and fibrogenesis in chronic diseases. Understanding these mechanisms may contribute to the development of new therapeutic strategies to clinically manage fibroproliferative diseases and organ-specific fibrotic disorders.

Thyroid surgery at a volunteer program in Sub-Saharan Africa. / Cirugía tiroidea en programas de cooperación en África subsahariana.

INTRODUCTION: The aim of this study is to demonstrate our experience at a volunteer surgical program in Cameroon, which is of special interest given to the inability to adopt international treatment guidelines for thyroid surgery in areas of limited resources due to the lack of preoperative testing and to the difficulty to obtain sustitutive hormonal treatment. METHODS: This is a prospective observational study that includes 16 cases of thyroid surgery in Dschang (Cameroon) during June 2015. The patients were previously selected by a local medical team. All patients were black, 15 women and one man, with a mean age of 41 years. The surgical technique used for the removal of unilateral disease was hemithyroidectomy with isthmectomy and bilateral subtotal thyroidectomy for bilateral disease. RESULTS: Five subtotal thyroidectomies, 9hemithyroidectomies and 2isthmectomies were performed. Prethyroid muscles were divided only in one case. We visualized 86% of the parathyroid glands and 84% of the recurrent laryngeal nerves. The main complications observed were one symptomatic cervical haematoma that required reoperation and 2surgical wound infections. There were no clinical episodes of hypocalemia or recurrent nerve lesion. The mean length of stay was 2.3 days. At follow-up, all bilateral thyroidectomies developed high TSH levels. CONCLUSIONS: Thyroid surgery is safe in developing countries adopting protocols and techniques we use in our environment (avoiding total thyroidectomy). Bilateral thyroidectomies should not be performed unless functional studies are available in the follow-up and a thyroid hormone supplement stock guaranteed whenever necessary.

Association between SNPs of Metalloproteinases and Prostaglandin F2α Receptor Genes and Latanoprost Response in Open-Angle Glaucoma.

PURPOSE: To determine whether single nucleotide polymorphisms (SNPs) of genes coding for matrix metalloproteinases (MMPs) and the prostaglandin F2α receptor gene (PTGFR) are related to a response to latanoprost in a white Spanish population of glaucomatous patients. DESIGN: Case-control study. PARTICIPANTS: One hundred twenty-four patients with open-angle glaucoma. METHODS: Genotyping was performed in 117 patients with primary open-angle glaucoma with a minimum treatment duration of 4 weeks. Candidate genes and individual polymorphisms were selected according to the effect on the mechanism of action of latanoprost. Multi-SNP haplotype analyses for associations also were tested. MAIN OUTCOME MEASURES: Diurnal intraocular pressure reduction and genotyping of the SNPs in the MMPs and PTGFR. RESULTS: The PTGFR SNPs were associated with positive (rs6686438, rs10786455) and negative (rs3753380, rs6672484, rs11578155) responses to latanoprost. Multiple testing found 2 genes, PTGFR and MMP-1, were related to refractoriness to latanoprost. CONCLUSIONS: The SNPs of the PTGFR and MMP-1 genes may determine the latanoprost response in a white European Spanish population. This study identified 5 SNPs related to the latanoprost response; 1 SNP, rs3753380, already has been associated with a poor response to latanoprost in a healthy Japanese population. Latanoprost is a commonly used antiglaucomatous drug, and increased knowledge of its mechanism of action will lead to advances in pharmacogenetics.

Parenteral solution of nutritional hepatotrophic factors improves regeneration in thioacetamide-induced cirrhotic livers after partial hepatectomy.

Liver resection is a suitable option for the treatment of certain hepatic conditions, particularly hepatocarcinomas, in patients with cirrhosis. However, this disease impairs liver regeneration, which increases the risk of liver failure and postoperative death. Supportive treatments for regeneration of the remaining liver may be useful for the recovery of these patients. We demonstrated that nutritional hepatotrophic factors (NHF) is an effective regenerative stimulus for cirrhotic livers in rats subjected to partial hepatectomy (PH). The rats with thioacetamide-induced cirrhosis were subjected to PH, and they were divided into 2 groups. One group received intraperitoneal administration of NHF, and the other group received saline solution. After 12 days, biometric data, collagen content, hepatocyte regeneration (proliferation cell nuclear antigen immunochemistry), and profibrotic gene expression (Collagen-α1, matrix metalloproteinase 2, tissue inhibitor of metalloproteinase 1, and transforming growth factor beta 1) were assessed. The results indicated that the rats treated with NHF after PH had an increased liver size, a reduced amount of collagen, and a higher hepatocyte proliferation index compared with the rats that underwent PH alone. In addition, collagen-α1 gene expression was decreased in the NHF-treated rats. Thus, postoperative improvement in the liver morphology following NHF treatment may cause a significant decrease in the risk of liver failure and mortality after hepatic resection.

Bioethical knowledge in students and health professionals: a systematic review.

Introduction: Bioethics training is essential for healthcare professionals as it enables them to address ethical dilemmas in their clinical practice. However, there is still a lack of rigorous teaching programs, and assessing bioethical knowledge poses challenges. Methodology: Systematic review using the PRISMA method. Results: Analysis of 27 studies reveals a lack of ethical knowledge and skills among healthcare professionals and students. Specific training in bioethics is effective in developing bioethical competencies. Different approaches have been employed, including integrated training in academic curricula and intensive or ongoing programs. The results demonstrate improvements in knowledge, attitudes, and ethical values, although regularly updating these courses is recommended. Conclusion: Specific training, institutional support, and considering regional and disciplinary differences are necessary to enhance ethics in the practice of healthcare professionals. Systematic review registration: https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42023437146, identifier CRD42023437146.

Relationships between experiences of humiliation on social networks, problematic phone use, and aggressive and altruistic behaviors in young adults.

The purpose of this study was to analyze the relationships between cybervictimization in social networks, problematic smartphone use, aggressive behaviors, and prosocial altruistic tendencies in young adults. The sample consisted of 601 young adults (mean age = 19.96 years; SD = 2.27; 69.1% female) who were administered online assessments of experiences of humiliation on networks, problematic smartphone use, prosocial altruistic tendencies, and aggressiveness. Results indicated significant indirect effects of cyber victimization on aggressiveness and prosocial altruistic tendencies through problematic smartphone use. Problematic cell phone use explained the relationships between online humiliation and aggressive and prosocial altruistic behaviors. The results confirmed the positive relationship between cybervictimization and problematic cell phone use, consistent with previous research. However, the negative relationship between cybervictimization and altruistic prosocial tendencies was not corroborated. The findings emphasize the need to promote actions that foster social connectedness and interdependence among young individuals to develop their identity within the community.

Evaluation of Thermography as a Diagnostic Technique in Asymptomatic or Incipient Onychomycosis.

Onychomycosis is usually diagnosed symptomatically due to the very clear signs caused by the fungus on the nail surface and structure, although the growth of the infecting agent must also be verified by culture in an enriched medium. This procedure is normally lengthy (four weeks), and samples can be contaminated, delaying the prescription of appropriate and effective treatment. Only one previous study has addressed the possibility of using thermography as a diagnostic method for onychomycosis in older people (31-70 years). The present study confirms this use but in individuals aged 18-31 years with incipient mycosis and no pathological signs. Using an FLIR E60 BX camera in a study with 214 samples, we found that men had more onychomycosis than women. We observed a relation between the presence of infection and nail temperature, with a higher temperature in yeast infections (+1 °C) and a lower temperature in dermatophyte infections (-2 °C). A higher temperature by almost 1 °C was also observed in older participants. Thermography can be viewed as a new diagnostic method in asymptomatic or incipient onychomycosis, providing the thermographic camera is sufficiently sensitive and the appropriate procedure is followed, although fungal culture is always necessary to confirm recovery after treatment.

Intravitreal allogeneic mesenchymal stem cells: a non-randomized phase II clinical trial for acute non-arteritic optic neuropathy.

BACKGROUND: An effective treatment for acute non-arteritic ischemic optic neuropathy (NA-AION) has not been known or proven yet. Previous studies have suggested a neuroprotective effect of allogeneic bone marrow-derived mesenchymal stem cells. This study aims to report the results of a clinical trial on patients with acute non-arteritic optic neuropathy (NA-AION) treated with an intravitreal injection of allogeneic bone marrow-derived mesenchymal stem cells (BM-MSCs) (MSV®). METHODS: We conducted a prospective, non-randomized, clinical phase-II study (Eudra CT number 2016-003029-40; ClinicalTrials.gov Registry NCT03173638) that included 5 patients with acute unilateral NA-AION diagnosed within 2 weeks after symptom onset and who received an intravitreal injection of allogeneic BM-MSCs (0.05 ml; cell concentration: 1.5 × 106cells/mL). The patients underwent regular ophthalmological examinations and were followed for one year. RESULTS: In this trial, allogeneic BM-MSCs appeared to be safe as no patients developed signs of acute nor chronic intraocular inflammation or a significant change in intraocular pressure, although an epiretinal membrane was developed in one patient. A retrolental aggregate formed shortly after the injection spontaneously disappeared within a few weeks in another phakic patient, leaving a subcapsular cataract. Visual improvement was noted in 4 patients, and amplitudes of P100 on the visually evoked potentials recordings increased in three patients. The retinal nerve fiber layer and macular ganglion cell layer thicknesses significantly decreased during the follow-up. CONCLUSIONS: Besides the development of an epiretinal membrane in one patient, the intravitreal application of allogeneic BM-MSCs appeared to be intraocularly well tolerated. Consequently, not only NA-AION but also BM-MSCs deserve more clinical investigational resources and a larger randomized multicenter trial that would provide stronger evidence both about safety and the potential therapeutic efficacy of intravitreally injected allogeneic BM-MSCs in acute NA-AION. TRIAL REGISTRATION: Safety Assessment of Intravitreal Mesenchymal Stem Cells for Acute Non-Arteritic Anterior Ischemic Optic Neuropathy (NEUROSTEM). NCT03173638. Registered June 02, 2017 https://clinicaltrials.gov/ct2/show/NCT03173638 .

Review on Treatment Planning Systems for Cervix Brachytherapy (Interventional Radiotherapy): Some Desirable and Convenient Practical Aspects to Be Implemented from Radiation Oncologist and Medical Physics Perspectives.

Intracavitary brachytherapy (BT, Interventional Radiotherapy, IRT), plays an essential role in the curative intent of locally advanced cervical cancer, for which the conventional approach involves external beam radiotherapy with concurrent chemotherapy followed by BT. This work aims to review the different methodologies used by commercially available treatment planning systems (TPSs) in exclusive magnetic resonance imaging-based (MRI) cervix BT with interstitial component treatments. Practical aspects and improvements to be implemented into the TPSs are discussed. This review is based on the clinical expertise of a group of radiation oncologists and medical physicists and on interactive demos provided by the software manufacturers. The TPS versions considered include all the new tools currently in development for future commercial releases. The specialists from the supplier companies were asked to propose solutions to some of the challenges often encountered in a clinical environment through a questionnaire. The results include not only such answers but also comments by the authors that, in their opinion, could help solve the challenges covered in these questions. This study summarizes the possibilities offered nowadays by commercial TPSs, highlighting the absence of some useful tools that would notably improve the planning of MR-based interstitial component cervix brachytherapy.

Development of real-time PCR methods for the quantification of Methanoculleus, Methanosarcina and Methanobacterium in anaerobic digestion.

Anaerobic digestion is a growing technology to manage organic waste and produce bioenergy. To promote this technology, it is essential to know, at the molecular level, the dynamics of microbial communities, specifically the methanogenic community. In the present study, three primer pairs were selected from seven primer pairs which were designed and tested with different concentrations and conditions to detect Methanosarcina, Methanoculleus and Methanobacterium by real-time PCR based on the SYBR Green System. The functionality of the developed methods was demonstrated by the high linear relationship of the standard curves, and the specificity of each primer was empirically verified by testing DNA isolated from methane-producing and non-producing strains. These assays also exhibited good repeatability and reproducibility, which indicates the robustness of the methods. The described primers were successfully used to investigate the methanogenic communities of 10 samples from an anaerobic co-digestion. The genus Methanosarcina was the dominant methanogenic group.

Advanced Therapy Medicinal Products for the Eye: Definitions and Regulatory Framework.

Advanced therapy medicinal products (ATMPs) are a group of innovative and complex biological products for human use that comprises somatic cell therapy medicinal products, tissue engineered products, gene therapy medicinal products, and the so-called combined ATMPs that consist of one of the previous three categories combined with one or more medical devices. During the last few years, the development of ATMPs for the treatment of eye diseases has become a fast-growing field as it offers the potential to find novel therapeutic approaches for treating pathologies that today have no cure or are just subjected to symptomatic treatments. Therefore, it is important for all professionals working in this field to be familiar with the regulatory principles associated with these types of innovative products. In this review, we outline the legal framework that regulates the development of ATMPs in the European Union and other international jurisdictions, and the criteria that each type of ATMP must meet to be classified as such. To illustrate each legal definition, ATMPs that have already completed the research and development stages and that are currently used for the treatment of eye diseases are presented as examples.

Effects of Alpha-Connexin Carboxyl-Terminal Peptide (aCT1) and Bowman-Birk Protease Inhibitor (BBI) on Canine Oral Mucosal Melanoma (OMM) Cells.

Oral mucosal melanomas (OMM) are aggressive cancers in dogs, and are good models for human OMM. Gap junctions are composed of connexin units, which may have altered expression patterns and/or subcellular localization in cancer cells. Cell-to-cell communication by gap junctions is often impaired in cancer cells, including in melanomas. Meanwhile, the upregulated expression of the gap junction protein connexin 43 (Cx43) inhibits melanoma progression. The α-connexin carboxyl-terminal (aCT1) peptide reportedly maintains Cx43 expression and cell-cell communication in human mammary cells and increases the communication activity through gap junctions in functional assays, therefore causing decreased cell proliferation. The Bowman-Birk protease inhibitor (BBI), a component of soybeans, induces Cx43 expression in several tumor cells as a trypsin-chymotrypsin inhibition function, with antineoplastic effects. This study investigated the effect of aCT1 peptide and BBI treatment, alone or in combination, on TLM1 canine melanoma cell viability. Cell viability after treatment with aCT1, the reverse sequence peptide (R-pep), and/or BBI for 5 days was analyzed by PrestoBlue assay. Immunofluorescence was used to observe Cx43 localization and expression. aCT1 (200 µM) alone did not significantly decrease cell viability in TLM1 cells, whereas BBI (400 µg/ml) alone significantly decreased the TLM1 viability. Combined treatment with both aCT1 (200 µM) and BBI (400 µg/ml) significantly decreased cell viability in TLM1 cells. Cx43 expression, as identified by immunostainings in TLM1 cells, was increased in the cell membrane after the combination treatment with BBI and aCT1. This dual treatment can be combined to achieve the anticancer activity, possibly by increasing Cx 43 expression and affecting Cx43 migration to the cell membrane. In conclusion, a treatment strategy targeting Cx43 with BBI and aCT1 may possibly lead to new effective therapies for canine OMM.

Quantification of Global DNA Methylation in Canine Melanotic and Amelanotic Oral Mucosal Melanomas and Peripheral Blood Leukocytes From the Same Patients With OMM: First Study.

Oral mucosal melanomas (OMMs) are aggressive and resistant cancers of high importance in veterinary oncology. Amelanotic OMM produces comparatively less melanin and is considered to be more aggressive than melanotic OMM. Global DNA methylation profiles with hypomethylated or hypermethylated patterns have both been associated with aggressive neoplasms; however, global DNA hypomethylation seems to correlate to higher aggressiveness. Accordingly, global DNA methylation in peripheral blood leukocytes has been investigated to understand the role of systemic or environmental factors in cancer development. This study aimed to quantify global DNA methylation in canine melanotic and amelanotic OMM samples and in the peripheral blood leukocytes of the same dogs. Tumor tissue samples were collected from 38 dogs, of which 19 were melanotic and 19 were amelanotic OMM. These were submitted to immunohistochemistry (IHC) with anti-5-methylcytosine (5mC) and anti-Ki67 primary antibodies. Ki67- and 5mC-positive nuclei were manually scored with the help of an image analysis system. Peripheral blood samples were collected from 18 among the 38 OMM-bearing dogs and from 7 additional healthy control dogs. Peripheral blood leukocytes were isolated from the 25 dogs, and DNA was extracted and analyzed by high-performance liquid chromatography (HPLC) for global DNA methylation. The pattern of global DNA methylation in both canine melanotic and amelanotic OMM indicated higher percentages of weakly or negatively stained nuclei in most of the OMM cells, presuming predominant global DNA hypomethylation. In addition, Ki67 counts in amelanotic OMM were significantly higher than those in melanotic OMM (p < 0.001). Global DNA methylation different immunostaining patterns (strong, weak or negative) correlated with Ki67 scores. Global DNA methylation in circulating leukocytes did not differ between the 9 melanotic and 9 amelanotic OMM or between the 18 OMM-bearing dogs and the 7 healthy dogs. This study provides new information on canine melanotic and amelanotic OMM based on global DNA methylation and cell proliferation.

The skin structures and their role in the thermoregulation of the South American fur seal (Arctocephalus australis).

The skin of the South American fur seal (Arctocephalus australis) is important for animal thermoregulation in both terrestrial and aquatic environments. Skin tissue samples were collected from A. australis for microscopic analysis and were related to anatomical references. The aim of this study was to describe the skin morphology, as well as to suggest the major anatomical regions and skin components involved in the thermoregulation of this species. Using light microscopy, the skin of six animals was examined based on histological, morphometric, and immunohistochemical criteria. Hair follicle morphology was examined using scanning electron microscopy. The skin was classified as either thick or thin based on its epidermal thickness. The thin epidermis regions had more abundant hair follicles, as well as high pigmentation, whereas the thick epidermis regions had very pigmented epidermal layers. Pigmentation of hair and skin is fundamental for protection against ultraviolet rays; moreover, hair is important in preventing abrasion, and provides an insulating layer against the external environment, which can be much colder than body temperature. Furthermore, the dermis is well vascularized, especially the superficial dermis. All regions of the skin have adaptations for maintaining the animal's condition in both terrestrial and aquatic environments. Among the studied regions, the interdigital region from hindflipper showed important morphological characteristics related to thermoregulation, such as having an epidermis of intermediate thickness, a dermis with a small number of hairs, a large amount of blood vessels, and sweat glands with large lumens, indicating that heat exchange in this region may be faster.

Atrophy and neuron loss: effects of a protein-deficient diet on sympathetic neurons.

Protein deficiency is one of the biggest public health problems in the world, accounting for about 30-40% of hospital admissions in developing countries. Nutritional deficiencies lead to alterations in the peripheral nervous system and in the digestive system. Most studies have focused on the effects of protein-deficient diets on the enteric neurons, but not on sympathetic ganglia, which supply extrinsic sympathetic input to the digestive system. Hence, in this study, we investigated whether a protein-restricted diet would affect the quantitative structure of rat coeliac ganglion neurons. Five male Wistar rats (undernourished group) were given a pre- and postnatal hypoproteinic diet receiving 5% casein, whereas the nourished group (n = 5) was fed with 20% casein (normoproteinic diet). Blood tests were carried out on the animals, e.g., glucose, leptin, and triglyceride plasma concentrations. The main structural findings in this study were that a protein-deficient diet (5% casein) caused coeliac ganglion (78%) and coeliac ganglion neurons (24%) to atrophy and led to neuron loss (63%). Therefore, the fall in the total number of coeliac ganglion neurons in protein-restricted rats contrasts strongly with no neuron losses previously described for the enteric neurons of animals subjected to similar protein-restriction diets. Discrepancies between our figures and the data for enteric neurons (using very similar protein-restriction protocols) may be attributable to the counting method used. In light of this, further systematic investigations comparing 2-D and 3-D quantitative methods are warranted to provide even more advanced data on the effects that a protein-deficient diet may exert on sympathetic neurons. (c) 2009 Wiley-Liss, Inc.

Inhibitory effects of Euphorbia tirucalli latex on murine B16/F10 melanoma cells and lung metastasis.

Euphorbia tirucalli (E. tirucalli) is a tropical and subtropical plant that produces a latex which is used for several purposes. The components of E. tirucalli latex include triterpenes, diterpenes and steroids. The aim of the present study was to evaluate the effects of diluted E. tirucalli latex on murine B16/F10 melanoma cells and lung metastasis. For this purpose, an in vitro study was first performed, in which B16/F10 cells were treated with diluted (1/2 to 1/11,192) E. tirucalli latex. In a second study, B16/F10 melanoma cells were inoculated into the tail vein of mice to generate lung metastases; the mice then received 0.467 µg of latex diluted in 200 ml saline by gavage for 14 days. A significant decrease in B16/F10 cell viability was observed using the MTT assay at 24 and 48 h after treatment with E. tirucalli latex. In addition, a significant decrease in the volume fraction occupied by B16/F10 metastatic colonies in the lungs was observed in mice treated with E. tirucalli latex. These results confirm the antineoplastic effects of diluted E. tirucalli latex.