Cytopathology smears from autopsies: A viable storage method for molecular analysis.

INTRODUCTION: Cytology appears to be a viable option to histological samples for proper storage and maintenance of autopsy material for DNA extraction and analysis. In the present study, we tested the feasibility of using archived air-dried smears produced at the time of the autopsy for simple molecular analysis, comparing quantity and quality of the DNA extracted from the smears to that of correspondent histological specimens. METHODS: Air-dried cytological smears were obtained from scrapings of exactly the same areas collected for histological study. DNA was extracted using a commercially available protocol from all samples, with calculation of purity ratio and overall concentration. The integrity of the extracted DNA was also verified through conventional polymerase chain reaction (PCR). RESULTS: Five cases of lung tumours (2 small cell carcinomas and 3 adenocarcinomas) were collected. Percentage of tumour cells and necrosis ranged from 30% to 90% and from 10% to 40%, respectively, in the cytological preparations, and from 50% to 90% and from 10% to 80%, respectively, in the histological preparations. Purity ratio (260/280) had a median of 1.87 in cytology vs 1.94 in histology. Mean DNA concentration among the cytological preparations was 2653 ng/mL (range 1684-3980 ng/mL) vs 757.2 ng/mL among the histological preparations (range 456-1829 ng/mL. DNA from all five cases of cytology was successfully amplified by conventional PCR, in contrast to none from the histology specimens. CONCLUSIONS: Archived air-dried smears scraped from tumoural lesions in autopsies have proven to yield a good concentration of quality DNA for conventional PCR, with better results than formalin-fixed paraffin embedded material.

Entropy of Entanglement between Quantum Phases of a Three-Level Matter-Radiation Interaction Model.

We show that the entropy of entanglement is sensitive to the coherent quantum phase transition between normal and super-radiant regions of a system of a finite number of three-level atoms interacting in a dipolar approximation with a one-mode electromagnetic field. The atoms are treated as semi-distinguishable using different cooperation numbers and representations of SU(3), variables which are relevant to the sensitivity of the entropy with the transition. The results are computed for all three possible configurations ( Ξ , Λ and V) of the three-level atoms.

Geometry and Entanglement of Two-Qubit States in the Quantum Probabilistic Representation.

A new geometric representation of qubit and qutrit states based on probability simplexes is used to describe the separability and entanglement properties of density matrices of two qubits. The Peres-Horodecki positive partial transpose (ppt) -criterion and the concurrence inequalities are formulated as the conditions that the introduced probability distributions must satisfy to present entanglement. A four-level system, where one or two states are inaccessible, is considered as an example of applying the elaborated probability approach in an explicit form. The areas of three Triadas of Malevich's squares for entangled states of two qubits are defined through the qutrit state, and the critical values of the sum of their areas are calculated. We always find an interval for the sum of the square areas, which provides the possibility for an experimental checkup of the entanglement of the system in terms of the probabilities.

How to do it: Teaching surgical skills to medical undergraduates.

Medical students must be capable of performing clinical and surgical procedures in outpatient care and initial emergency care in all stages of the biological cycle. Here, we describe the surgical skills schedule with different animal models fulfilled at the Municipal University of São Caetano do Sul (USCS) Medical School, São Caetano do Sul, SP, Brazil, during the surgical abilities module. We retrospectively reviewed the surgical abilities module schedule provided at the USCS Medical School from 2015 until 2020; in this paper, we describe the use of different animal models. The activities were developed for two semesters during medical school and included an ox tongue, cylindrical Styrofoam, chicken leg and neck, live rabbits, and pigs. Practical surgical teaching starts with sutures using the ox tongue, after which students are taught to perform tenorrhaphy using cylindrical Styrofoam and chicken legs, followed by vascular anastomosis using the chicken trachea and esophagus. Rabbits are appropriate for training a variety of procedures such as cystostomy, gastrostomy, and appendectomy. Pigs allow for the simulation of several types of procedures such as chest drainage. Surgical training for medical undergraduates was demonstrated with an evolutionary intent, starting with simple sutures and ending up with basic emergency room surgical procedures.

Standardization of experimental model regarding star fruit intoxication in Wistar rats suffering with nephropathy.

PURPOSE: To present a model to reproduce the clinical condition, in order to better understand the pathophysiology of neurological impairment related to intoxication. METHODS: Twenty-five Wistar rats were used and divided into five groups: Shaw group (WHI), water gavage group (WGV), star fruit gavage group (SGV), nephropathic group with water gavage (NPW), nephropathic group with star fruit gavage (NPS).Nephropathic groups were submitted to surgery, developing nephropathy. After surgery, they received preestablished gavage with star fruit juice or water. The electroencephalographic records were evaluated in the experimental nephropathic group that received gavage of star fruit juice. RESULTS: To assess the induction of neurotoxicity using electroencephalographic data, the NPS group demonstrated the presence of epileptic seizures associated with star fruit intoxication. CONCLUSIONS: The experimental model herein presented was adequate to reproduce the clinical condition experienced by nephropathic patients who ingest star fruit juice, establishing, thus, an experimental model utterly important for the study of the neurological toxicity process.

C-Fos expression in epileptogenic areas of nephropathic rats undergoing star fruit poisoning.

Purpose Studies have demonstrated that star fruit consumption by nephropathic patients triggers severe neurotoxic effects that can lead to convulsions or even death. Brain areas likely susceptible to star fruit poisoning have not been investigated. The objective of the present study was to map possible epileptogenic areas susceptible to star fruit intoxication in nephropathic rats. Methods The study analyzed 25 rats (5 groups). Rats in the experimental group underwent bilateral ureteral obstruction surgery and orogastric gavages with star fruit juice. An electroencephalogram was used to confirm convulsive seizures. Urea and creatinine levels were used to confirm the uremia model. Immunohistochemical analysis was used to map cells with c-Fos protein (c-Fos+ cells) to identify brain areas with increased neuronal activity. Control groups included non-nephropathic and nephropathic rats that did not receive star fruit. Results A statistically significant increase (p<0.01) in c-Fos+ cells was noted in nephropathic animals receiving star fruit juice compared to control groups, in brain areas commonly related to epileptogenic neural circuits including the hippocampus, amygdala, rhinal cortex, anterior cingulate area, piriform area, and medial dorsal thalamus. Conclusion These data corroborate the neurotoxic capacity of star fruit in nephropathic patients.

Amitriptyline attenuates interstitial inflammation and ameliorates the progression of renal fibrosis.

Amitriptyline is a pleiotropic tricyclic antidepressant, which has anti-oxidant and anti-inflammatory properties. We tested whether amitriptyline might be useful in the treatment of chronic renal disease using the mouse model of unilateral ureteral obstruction. Amitriptyline caused a significant reduction of interstitial fibrosis, determined by Masson's staining, with minimal myofibroblast formation and macrophage infiltration following ureteral obstruction. Using quantitative PCR we found that this treatment significantly reduced the expression of key molecular markers of progressive tubulointerstitial injury such as osteopontin, MCP-1, ICAM-1, and TGF-beta1 compared to their level in a saline-treated control group. Sublethal X-irradiation or mycophenolate mofetil, treatments that reduce inflammation, were comparable to amitriptyline in the reduction of interstitial fibrosis and macrophage infiltration. These studies in animals suggest that amitriptyline is worth testing as a therapeutic agent that might preserve renal function by blocking inflammation and renal fibrosis.

Standardization of experimental model regarding star fruit intoxication in Wistar rats suffering with nephropathy

ABSTRACT Purpose To present a model to reproduce the clinical condition, in order to better understand the pathophysiology of neurological impairment related to intoxication. Methods Twenty-five Wistar rats were used and divided into five groups: Shaw group (WHI), water gavage group (WGV), star fruit gavage group (SGV), nephropathic group with water gavage (NPW), nephropathic group with star fruit gavage (NPS).Nephropathic groups were submitted to surgery, developing nephropathy. After surgery, they received preestablished gavage with star fruit juice or water. The electroencephalographic records were evaluated in the experimental nephropathic group that received gavage of star fruit juice. Results To assess the induction of neurotoxicity using electroencephalographic data, the NPS group demonstrated the presence of epileptic seizures associated with star fruit intoxication. Conclusions The experimental model herein presented was adequate to reproduce the clinical condition experienced by nephropathic patients who ingest star fruit juice, establishing, thus, an experimental model utterly important for the study of the neurological toxicity process.

C-Fos expression in epileptogenic areas of nephropathic rats undergoing star fruit poisoning

Abstract Purpose Studies have demonstrated that star fruit consumption by nephropathic patients triggers severe neurotoxic effects that can lead to convulsions or even death. Brain areas likely susceptible to star fruit poisoning have not been investigated. The objective of the present study was to map possible epileptogenic areas susceptible to star fruit intoxication in nephropathic rats. Methods The study analyzed 25 rats (5 groups). Rats in the experimental group underwent bilateral ureteral obstruction surgery and orogastric gavages with star fruit juice. An electroencephalogram was used to confirm convulsive seizures. Urea and creatinine levels were used to confirm the uremia model. Immunohistochemical analysis was used to map cells with c-Fos protein (c-Fos+ cells) to identify brain areas with increased neuronal activity. Control groups included non-nephropathic and nephropathic rats that did not receive star fruit. Results A statistically significant increase (p<0.01) in c-Fos+ cells was noted in nephropathic animals receiving star fruit juice compared to control groups, in brain areas commonly related to epileptogenic neural circuits including the hippocampus, amygdala, rhinal cortex, anterior cingulate area, piriform area, and medial dorsal thalamus. Conclusion These data corroborate the neurotoxic capacity of star fruit in nephropathic patients.

Use of insulin-like growth factor in the healing of open wounds in diabetic and non-diabetic rats.

PURPOSE: To analyze the effects of application of 1% and 3% insulin-like growth factor I (IGF-1) cream on the process of wound healing in induced skin lesions in diabetic and non-diabetic rats and evaluate its effect on expression of myofibroblasts. METHODS: Ninety-six Wistar adult male rats were divided into six groups, with 16 rats in each group, as follows: group 1: non-diabetic, untreated; group 2: non-diabetic, treated with 1% IGF-1 cream; group 3: non-diabetic, treated with 3% IGF-1 cream; group 4: diabetic, untreated; group 5: diabetic, treated with 1% IGF-1 cream; and group 6: diabetic, treated with 3% IGF-1 cream. In groups 4, 5, and 6, diabetes was induced by intravenous injection of alloxan. After diabetes had been induced, animals were mantained for 3 months. The experimental procedure consisted of the creation of a circular incision of 0.9 mm in diameter using a metal punch. Following this, wounds were treated daily according to the assigned treatment regimen. Groups 2 and 5 were treated with 1% IGF-1 cream, groups 3 and 6 with 3% IGF-1 cream, and groups 1 and 4 and the untreated groups with 0.9% saline solution. From each group, samples from 4 rats were taken at three, seven, 14, and 21 days after the injury. Samples were fixed in 10% formalin to prepare slides for histological analysis. Slides stained with hematoxylin-eosin (H&E) and Masson were observed vascular proliferation, mononuclear cells, polymorphonuclear cells, fibroblast proliferation, re-epithelialization, and collagen fibers. This study analyzed the expression of α-smooth muscle actin using specific antibodies to correlate the temporal expression of α-smooth muscle-specific actin (α-SM actin), a molecular marker for myofibroblast transformation. RESULTS: Macroscopic observation of wounds showed a more rapid re-epithelialization of wounds treated with IGF. Regarding acute inflammatory reactions, the results of the analysis of vascular proliferation and polymorphonuclear and mononuclear cells showed no statistically significant differences in any of the periods studied (according to the results of a Mann-Whitney test). The initial immunohistochemical analysis of tissue samples conducted to compare the expression of α-smooth muscle actin between groups showed a relevant response in the expression of myofibroblasts. Data were analyzed using ANOVA and were found to be statistically significant. CONCLUSION: The topical application of 1% and 3% IGF-1 creams increases the expression of myofibroblasts in the process of wound healing in rats.

Application of high frequency bipolar electrocoagulation LigaSure™ in appendix vermiformis of rabbits with or without acute inflammatory process.

PURPOSE: To evaluate the efficacy of the use of LigaSure™ in appendectomy, with or without acute inflammatory process, and to compare with simple ligature and conventional therapy. METHODS: A total of 30 rabbits (Oryctolagus cuniculus) randomly allocated in two groups, group A and B, of 15 animals each were used. The group A without acute appendicitis and the group B with acute appendicitis were submitted to appendectomy. After, the groups were subdivided into three groups, each group containing five rabbits submitted to simple ligature, conventional therapy and application of LigaSure™. We assessed macroscopic and microscopy parameters of appendiceal stump and operative wound. RESULTS: The group with acute appendicitis that LigaSure™ was applied had fibrosis in 100% of animals, as well as in the other operative techniques used. It suggested that application of LigaSure™ is efficient as other techniques used in healing of appendiceal stump. CONCLUSIONS: The application of LigaSure™ induces the formation of fibrosis in the appendiceal stump. The technique proved efficacy to induce enough fibrous tissue to obstruct leakage of enteric content.

Experimental model for learning in vascular surgery and microsurgery: esophagus and trachea of chicken.

PURPOSE: This paper proposes a model of training surgical skills using vascular anastomosis in an animal model that simulates the size, consistency and resistance arteries and veins, to use it to chicken trachea and esophagus, respectively. METHODS: We used chicken necks where the esophagus and trachea were dissected and after preparation were followed every step of the procedure of vascular anastomosis. The flow of the anastomosis was confirmed by direct observation and testing of filling (empty-and-refill test) immediately after the anastomosis. RESULTS: All samples proved to be viable by the criteria described above. CONCLUSION: For the first time presents an interesting experimental model used to train vascular sutures, because it is endowed with all the necessary requirements for the learning of experimental vascular surgery.

Use of insulin-like growth factor in the healing of open wounds in diabetic and non-diabetic rats

To analyze the effects of application of 1% and 3% insulin-like growth factor I (IGF-1) cream on the process of wound healing in induced skin lesions in diabetic and non-diabetic rats and evaluate its effect on expression of myofibroblasts. METHODS: Ninety-six Wistar adult male rats were divided into six groups, with 16 rats in each group, as follows: group 1: non-diabetic, untreated; group 2: non-diabetic, treated with 1% IGF-1 cream; group 3: non-diabetic, treated with 3% IGF-1 cream; group 4: diabetic, untreated; group 5: diabetic, treated with 1% IGF-1 cream; and group 6: diabetic, treated with 3% IGF-1 cream. In groups 4, 5, and 6, diabetes was induced by intravenous injection of alloxan. After diabetes had been induced, animals were mantained for 3 months. The experimental procedure consisted of the creation of a circular incision of 0.9 mm in diameter using a metal punch. Following this, wounds were treated daily according to the assigned treatment regimen. Groups 2 and 5 were treated with 1% IGF-1 cream, groups 3 and 6 with 3% IGF-1 cream, and groups 1 and 4 and the untreated groups with 0.9% saline solution. From each group, samples from 4 rats were taken at three, seven, 14, and 21 days after the injury. Samples were fixed in 10% formalin to prepare slides for histological analysis. Slides stained with hematoxylin-eosin (H&E) and Masson were observed vascular proliferation, mononuclear cells, polymorphonuclear cells, fibroblast proliferation, re-epithelialization, and collagen fibers. This study analyzed the expression of α-smooth muscle actin using specific antibodies to correlate the temporal expression of α-smooth muscle-specific actin (α-SM actin), a molecular marker for myofibroblast transformation. RESULTS: Macroscopic observation of wounds showed a more rapid re-epithelialization of wounds treated with IGF. Regarding acute inflammatory reactions, the results of the analysis of vascular.

Application of high frequency bipolar electrocoagulation LigaSureTM in appendix vermiformis of rabbits with or without acute inflammatory process / Aplicação do eletrocoagulador bipolar de alta frequência LigaSureTM no apêndice vermiforme na vigência ou não de processo inflamatório agudo

PURPOSE: To evaluate the efficacy of the use of LigaSureTM in appendectomy, with or without acute inflammatory process, and to compare with simple ligature and conventional therapy. METHODS: A total of 30 rabbits (Oryctolagus cuniculus) randomly allocated in two groups, group A and B, of 15 animals each were used. The group A without acute appendicitis and the group B with acute appendicitis were submitted to appendectomy. After, the groups were subdivided into three groups, each group containing five rabbits submitted to simple ligature, conventional therapy and application of LigaSureTM. We assessed macroscopic and microscopy parameters of appendiceal stump and operative wound. RESULTS: The group with acute appendicitis that LigaSureTM was applied had fibrosis in 100% of animals, as well as in the other operative techniques used. It suggested that application of LigaSureTM is efficient as other techniques used in healing of appendiceal stump. CONCLUSIONS: The application of LigaSureTM induces the formation of fibrosis in the appendiceal stump. The technique proved efficacy to induce enough fibrous tissue to obstruct leakage of enteric content.

OBJETIVO: Avaliar a eficácia da utilização do LigaSureTM na apendicectomia, com ou sem a presença de processo inflamatório agudo, comparando com ligadura simples ou técnica tradicional. MÉTODOS: Um total de 30 coelhos (Oryctolagus cuniculus) foi alocado em dois grupos, grupos A e B, cada um composto por 15 animais. O grupo A não apresentava apendicite aguda e o grupo B com apendicite aguda, sendo os animais submetidos à apendicitectomia. Cada grupo foi divido em três subgrupos, cada um com cinco animais onde foram então submetidos à ligadura simples, técnica tradicional ou utilização do LigaSureTM. Foram avaliados parâmetros macroscópicos assim como microscópicos do coto apendicular assim como da ferida operatória. RESULTADOS: No grupo que apresentava apendicite aguda onde foi utilizado o LigaSureTM, foi observado fibrose em 100% dos animais, assim como nos outros dois grupos. Com isto mostrou-se que a aplicação do LigaSureTM é tão eficiente quanto as outras técnicas utilizadas na cicatrizaçãoo do coto apendicular. CONCLUSÕES: A utilização do LigaSureTM induz a formação de fibrose na região do coto apencicular. A técnica foi eficaz quanto à formação de tecido fibrótico evitando o extravassamento de conteúdo entérico.

Estudo comparativo das complicações vasculares arteriais e biliares no transplante hepático pediátrico com doador vivo e cadavérico / Comparative study of arterial and biliary complications in pediatric liver transplantation with living and cadaveric donors

Introdução: O transplante de fígado melhorar significativamente a taxa de sobrevivência de crianças e adolescentes com doença hepática terminal. Os pacientes, uma vez que tinha um prognóstico fatal, hoje podem ser submetidos a transplante de fígado (TF), com taxas de sobrevida de aproximadamente 90% em um ano. A atresia biliar é a principal indicação para a população pediátrica e para aqueles sofreram uma portoenterostomia e não obtiveram bons resultados com a mesma, representando mais de 50% das crianças menores de 2 anos na Europa e nos EUA. O transplante representa a única opção restante de sobrevivência com um bom benefício terapêutico. Os avanços nos cuidados cirúrgicos e clínicos, levaram a melhoria das técnicas para aumentar a sobrevida dos pacientes submetidos a transplante de fígado. Porém, a hemorragia e as complicações biliares representam o tendão de Aquiles do desenvolvimento deste processo. Além disso, a escassez de órgãos para a população pediátrica motiva o desenvolvimento de novas modalidades de enxertos de fígado, como a redução do enxerto de fígado, o transplante de fígado dividido, e mais recentemente o de doadores vivos. As complicações arteriais e biliares continuam a ser uma importante causa de morbidade, mortalidade e perda do enxerto após o transplante...

Introduction: Liver transplantation greatly improved the survival rate of children and adolescents with terminal liver diseases. Patients, who once had a fatal prognosis, today can be submitted to liver transplantation (LxT), with survival rates of approximately 90% in one year. The biliary atresia is the main indication in the pediatric population and for those for whom underwent a portoenterostomy and did not obtain good results with it, representing more than 50% of children under the age of 2 years in Europe and the U.S, transplantation represents the only remaining therapeutic option with good survival benefit. Advances in clinical care and surgical techniques led to improved survival of patients undergoing liver transplantation, but still the arterial and biliary complications represent the Achilles tendon in the development of this procedure. Moreover, the shortage of organs for the pediatric population motivate the development of new modalities of liver grafts, such as reduced liver graft, the split-liver transplantation and more recently the living donors. Despite the improved results, vascular arterial and biliary remain an important cause of morbidity, mortality and graft loss after transplantation. Methods: Survey in Pubmed and Lilacs to gather data related complications of pediatric liver transplantation between 1999 to 2009...

Modelo para o aprendizado experimental em cirurgia e microcirurgia vascular: esôfago e traquéia de frango / Experimental model for learning in vascular surgery and microsurgery: esophagus and trachea of chicken

PURPOSE: This paper proposes a model of training surgical skills using vascular anastomosis in an animal model that simulates the size, consistency and resistance arteries and veins, to use it to chicken trachea and esophagus, respectively. METHODS: We used chicken necks where the esophagus and trachea were dissected and after preparation were followed every step of the procedure of vascular anastomosis. The flow of the anastomosis was confirmed by direct observation and testing of filling (empty-and-refill test) immediately after the anastomosis. RESULTS: All samples proved to be viable by the criteria described above. CONCLUSION: For the first time presents an interesting experimental model used to train vascular sutures, because it is endowed with all the necessary requirements for the learning of experimental vascular surgery.

OBJETIVO: O presente trabalho propõe um modelo de treinamento de habilidades cirúrgicas em anastomoses vasculares utilizando um modelo que simula o calibre, a consistência e a resistência arterial e venosa, utilizando-se para isto a traquéia e esôfago de frango, respectivamente. MÉTODOS: Foram utilizados pescoços de frango, onde foram dissecados esôfago e traquéia e após preparo foram seguidos todos os passos do procedimento da anastomose vascular. O fluxo das anastomoses foi comprovado por observação direta e teste de enchimento imediatamente após as anastomoses. RESULTADOS: Todas as amostras mostraram-se viáveis pelos critérios acima descritos. CONCLUSÃO: Pela primeira vez apresenta-se um modelo experimental interessante que serve para treinar suturas vasculares, pois é dotado de todos os requisitos necessários para o aprendizado da cirurgia vascular experimental.

Amitriptyline eliminates calculi through urinary tract smooth muscle relaxation.

BACKGROUND: We investigated the effects of amitriptyline in the urinary tract smooth muscle and urolithiasis. METHODS: Cats presenting with obstructive acute renal failure (ARF) received amitriptyline, and renal function and survival rates were analyzed. Isometric contractions and membrane potentials of rat, pig, or human isolated urinary tract smooth muscle were recorded in the presence or absence of amitriptyline. RESULTS: Twenty cats with obstructive ARF caused by urethral plugs received amitriptyline. In all cases, plugs were completely eliminated, and renal function returned to normal, with a 100% survival rate in the follow-up. Amitriptyline produced potent relaxations in rat urethral strips, accompanied by significant reductions in urethral ring membrane potential. This effect was prevented by pretreatment of urethral rings with 4-aminopyridine (4-AP), a voltage-dependent potassium channel blocker. Amitriptyline abolished in a reversible manner acetylcholine-, bradykinin-, and KCl-induced contractions in rat isolated bladder, and this effect was also prevented by 4-AP. Of interest, spontaneous and KCl-induced contractions of pig and human isolated ureter were also blocked by amitriptyline. CONCLUSION: Our results indicate that amitriptyline is an effective and potent relaxant of urinary tract smooth muscle and this effect is mediated by opening of voltage dependent-potassium channels. We suggest that amitriptyline administration may help to promote elimination of urinary calculi.