Implementação de um sistema de eletroforese capilar com detecção de fluorescência induzida por laser

A capillary electrophoresis system using laser induced fluorescence detection is described. A Raman system equipped with a microscope has been used to focus the laser beam on the capillary giving a lateral resolution of 1.5 mm. The fluorescence signal of the analyte (ZnPcTS - tetrasulfonated zinc-phthalocyanine) was collected by the microscope objectives and analysed by a monochromator with confocal characteristics equipped with a CCD detector. Electropherograms obtained with this system were compared to those obtained on a commercial instrument, showing that the described system presents a lower detection limit and better resolution.

Diamond synthesis by lasers: recent progress

This paper provides an up-to-date critical review of methods for diamond synthesis by lasers. A provisional classification of synthesizing methods is carried out, in order to present a varied and heterogeneous experimental work that is as clear as possible. Laser diamond CVD methods involve chemical processes in a carbon-containing system induced by specific absorption of laser radiation, while diamond PVD methods involve physical processes induced by an intense, highly-directed laser beam. Different methods involving coupled, simultaneous action of lasers and classic CVD agents are suggested and discussed as prospective approaches for diamond synthesis.

Avaliação da voz em pacientes submetidos à cordectomia com laser de CO2

OBJETIVO: Avaliar a voz de pacientes portadores de carcinomas iniciais glóticos submetidos à cordectomia com laser de CO2. MÉTODO: Foram avaliados 15 pacientes com diagnóstico de carcinoma espinocelular Tis e T1 glóticos. A avaliação foi feita por meio da análise perceptivo auditiva da voz, pela análise acústica computadorizada e videolaringoestroboscópica. Os pacientes responderam a um questionário de avaliação geral da voz e um protocolo de qualidade de vida relacionado à voz (QVV). Os resultados foram comparados aos de um grupo controle de indivíduos com laringes normais. RESULTADOS: A análise perceptivo-auditiva da voz revelou que a maioria dos pacientes submetidos à cordectomia apresentou algum grau de disfonia, à custa de rouquidão e soprosidade. Considerando os parâmetros acústicos analisados e os valores do grupo controle, observou-se a tendência de um pequeno incremento da freqüência fundamental, mas sem diferença estatisticamente significante; os valores de jitter, shimmer e da proporção harmônico-ruído apresentaram-se significantemente alterados. Os aspectos analisados na videolaringoestroboscopia mostraram-se melhores nas cordectomias menos extensas. Os escores do QVV sugerem que os pacientes tiveram um discreto impacto na qualidade de vida relacionada à voz. CONCLUSÕES: Apesar da presença de alterações na qualidade vocal dos pacientes submetidos à cordectomia com laser de CO2, os resultados funcionais tendem a ser bem aceitos pelos pacientes, com discreta repercussão na qualidade de vida.

Cicatrização da membrana timpânica na timpanocentese com laser de argônio comparado à técnica com microlanceta: estudo experimental em ratos

Otite média secretora (OMS) e otite média aguda recorrente (OMAR) podem necessitar tratamento cirúrgico para adequada ventilação da orelha média. A abertura clássica do tímpano (timpanocentese) requer incisão por microlanceta sob controle de microscópio cirúrgico e mantém-se patente por alguns dias. Estudos recentes sugerem que a timpanocentese feita por diferentes lasers pode permanecer permeável por maior tempo, o que possibilitaria a normalização da otite média. MATERIAL E MÉTODOS: Neste estudo experimental 34 ratos linhagem Wistar, albinos, machos adultos pesando cerca de 300g, foram anestesiados com cetamina 27 mg/kg e xilazina 2,7 mg/kg. A seguir, foram submetidos à timpanocentese incisional com microlanceta no ouvido direito (ML-OD), e à timpanocentese mediada por laser de argônio no ouvido esquerdo (LA-OE). RESULTADOS: Não houve diferença significativa no tempo de cicatrização das timpanocenteses feitas com laser de argônio ou microlanceta. Todas as timpanocenteses cicatrizaram em 10 dias. CONCLUSÃO: A timpanocentese com laser de argônio apresentou patência e cicatrização semelhantes à técnica clássica com microlanceta realizada em ratos Wistar sem enfermidades de orelha média.

Impacto do laser de baixa intensidade na supressão de infecções pelos vírus Herpes simplex 1 e 2: estudo in vitro

O uso do laser de baixa intensidade na supressão de infecções pelos vírus Herpes simplex 1 e 2 foi avaliado após uma a cinco aplicações, sendo observada uma redução gradual na replicação dos vírus Herpes simplex 1 e 2 com 68,4% e 57,3% de inibição, respectivamente, após 5 aplicações, indicando o seu uso clínico.

Espécimes de Schistosoma mansoni descritos por Pirajá da Silva no Brasil (1908) reexaminados pela técnica de microscopia de varredura a laser confocal

INTRODUÇÃO: Pirajá da Silva fez contribuição magnífica à helmintologia ao descrever ovos de Schistosoma mansoni nas fezes de um paciente, no Estado da Bahia e a morfologia de vermes adultos. MÉTODOS: Neste estudo, apresentamos uma avaliação microscópica das lâminas montadas e depositadas na Coleção Helmintológica do Instituto Oswaldo Cruz. A técnica empregada nesta nova análise foi a microscopia de varredura a laser confocal. RESULTADOS: Na parte anterior dos vermes adultos machos, observamos ventosas com musculatura bem desenvolvida e células germinativas dentro dos lobos testiculares. Visualizamos, também, espinhos localizados na região mediana do canal ginecóforo. Na superfície dorsal, encontramos tubérculos e feixes musculares transversais e longitudinais. Em relação ao aparelho reprodutivo feminino, pudemos distinguir um ovo no interior do útero e o ovário alongado com células germinativas. As glândulas vitelínicas estavam restritas à parte posterior das fêmeas conectadas por um ducto vitelínico curto. CONCLUSÕES: As características morfológicas são similares as estudadas anteriormente por Pirajá da Silva com vermes frescos. Além disso, este estudo demonstra a importância de se depositar espécimes nas coleções helmintológicas abrindo possibilidade de novos estudos com estas lâminas.

Recurrent herpes simplex infections: laser therapy as a potential tool for long-term successful treatment

Herpes simplex virus types 1 and 2 are the main infectious agents associated with oral and genital ulcerations. These infections are now widely recognized as sexually transmitted diseases. Among treatment options, low-level laser therapy (LLLT) has shown promising clinical results as a longer-lasting suppression therapy. Two clinical cases are described with recurrent labial herpes for which LLLT was used. Following treatment, both patients remained symptom free during the 17-month clinical follow-up period.

Transmyocardial laser revascularization. Early clinical experience

OBJECTIVE: To analyze the initial clinical experience of transmyocardial laser revascularization (TMLR) in patients with severe diffuse coronary artery disease. METHODS: Between February, 1998 and February, 1999, 20 patients were submitted to TMLR at the Heart Institute (InCor), University of São Paulo Medical School, Brazil, isolated or in association with conventional coronary artery bypass graft (CABG). All patients had severe diffuse coronary artery disease, with angina functional class III/IV (Canadian Cardiovascular Society score) unresponsive to medical therapy. Fourteen patients were submitted to TMLR as the sole therapy, whereas 6 underwent concomitant CABG. Fifty per cent of the patients had either been previously submitted to a CABG or to a percutaneous transluminal coronary angioplasty (PTCA). Mean age was 60 years, ranging from 45 to 74 years. RESULTS: All patients had three-vessel disease, with normal or mildly impaired left ventricular global function. Follow-up ranged from 1 to 13 months (mean 6.6 months), with no postoperative short or long term mortality. There was significant symptom improvement after the procedure, with 85% of the patients free of angina, and the remaining 15 % of the patients showing improvement in functional class, as well as in exercise tolerance. CONCLUSION: This novel technique can be considered a low risk alternative for a highly selected group of patients not suitable for conventional revascularization procedures.

In-stent restenosis. Acute and long-term outcomes after excimer laser coronary angioplasty

OBJETIVE: With the increased use of intracoronary stents, in-stent restenosis has become a clinically significant drawback in invasive cardiology. We retrospectively assessed the short- and long-term outcomes after excimer laser coronary angioplasty of in-stent restenosis. METHODS: Twenty-five patients with 33 incidents of in-stent restenosis treated with excimer laser coronary angioplasty (ELCA) were analyzed. Sixty-six percent were males, mean age of 73±11 years, and 83% were functional class III-IV (NYHA). ELCA was performed using 23 concentric and 10 eccentric catheters with a diameter of 1.6-2.2 mm, followed by balloon angioplasty (PTCA) and ultrasound monitoring. The procedure was performed in the following vessels: left anterior descending artery, 10; left circumflex artery, 8; right coronary artery, 6; left main coronary artery, 2; and venous bypass graft, 7. RESULTS: The ELCA was successful in 71% of the cases, and PTCA was 100% successful. The diameter of the treated vessels was 3.44±0.5mm; the minimal luminal diameter (MLD) increased from 0.30mm pre-ECLA to 1.97mm post-ELCA, and to 2.94mm post-PTCA (p<0.001). The percent stenosis was reduced from 91.4±9.5% before ECLA to 42.3±14.9% after ELCA and to 14.6 ± 9.3% after PTCA (p<0.001). Seventeen (68%) patients were asymptomatic at 6 months and 15 (60%) at 1 year. New restenosis rates were 8/33 (24.2%) at 6 months and 9 /33 (27.3%) at 12 months. CONCLUSION: ELCA is safe and effective for the treatment of in-stent restenosis. In the present sample, a slight increase in new restenotic lesions between 6 and 12 months was found.

Laser assisted extraction of pacemaker and implantable defibrillator leads

OBJECTIVE: To analyze the results of laser-assisted extraction of permanent pacemaker and defibrillator leads. METHODS: We operated upon 36 patients, whose mean age was 54.2 years, and extracted 56 leads. The reasons for extracting the leads were as follows: infection in 19 patients, elective replacement in 13, and other causes in 4 patients. The mean time of catheter placement was 7.5±5.5 years. Forty-seven leads were from pacemakers, and 9 were from defibrillators. Thirty-eight leads were in use, 14 had been abandoned in the pacemaker pocket, and 4 had been abandoned inside the venous system. RESULTS: We successfully extracted 54 catheters, obtaining a 96.4% rate of success and an 82.1% rate for complete extraction. The 2 unsuccessful cases were due to the presence of calcium in the trajectory of the lead. The mean duration of laser light application was 123.0±104.5 s, using 5,215.2±4,924.0 pulses, in a total of 24.4±24.2 cycles of application. Thirty-four leads were extracted from the myocardium with countertraction after complete progression of the laser sheath, 12 leads came loose during the progression of the laser sheath, and the remaining 10 were extracted with other maneuvers. One patient experienced cardiac tamponade after extraction of the defibrillator lead, requiring open emergency surgery. CONCLUSION: The use of the excimer laser allowed extraction of the leads with a 96% rate of success; it was not effective in 2 patients who had calcification on the lead. One patient (2.8%) had a complication that required cardiac surgery on an emergency basis.

Utilidade do Ultrassom intracardíaco no isolamento de veias pulmonares usando cateter-balão a laser

FUNDAMENTO: O isolamento das veias pulmonares (IVP) tem sido usado como endpoint para a ablação da fibrilação atrial (FA) com cateter balão. OBJETIVO: Determinar a utilidade do ultrassom intracardíaco (USIC) para guiar o IVP, usando cateter balão a laser. MÉTODOS: 59 VP foram ablacionadas em 27 cães. Imagens de Doppler foram usadas para identificar os vazamentos do fluxo sanguíneo entre a VP e o balão. Após cada liberação de energia, o cateter de mapeamento circular foi reposicionado para verificar se o isolamento tinha sido obtido. A posição de vazamento foi então correlacionada com a posição do gap no estudo patológico. A análise de regressão logística multivariada foi realizada. RESULTADOS: Cinquenta e nove VP foram submetidas à ablação. O tempo médio de energia liberada foi de 279±177 seg., o diâmetro médio do balão era de 23±3 mm, e o comprimento médio do balão era 25±4 mm. O isolamento completo foi obtido em 38/59 (64%), e foi significantemente mais comum sem vazamento: [30/38 (79%) versus 8/23 (35%), p<0,001]. Isso foi independente do tempo de aplicação (302±223 seg. vs. 266±148 sec., p=ns), potência (3,5 W/cm, 4,5 W/cm, e 5,5 W/cm), diâmetro do balão (24± 3 mm vs. 22± 3 mm, p= ns) e comprimento (27±4 mm vs. 24±4 mm, p=ns). O valor preditivo positivo para previsão de isolamento completo foi de 65% e o valor preditivo negativo foi 83%. CONCLUSÃO: Um vazamento identificável entre a VP e o dispositivo de ablação por cateter-balão observado no USIC é preditor de menor taxa de isolamento de VP mais baixas. O USIC pode ser útil para detectar vazamentos a fim de evitar o uso ineficaz de aplicação energia durante a ablação circunferencial da VP. Isto também pode ser útil quando outras energias são utilizadas.

Hemodynamic Effect of Laser Therapy in Spontaneously Hypertensive Rats

Systemic arterial hypertension (SAH) is considered to be the greatest risk factor for the development of neuro-cardiovascular pathologies, thus constituting a severe Public Health issue in the world. The Low-Level Laser Therapy (LLLT), or laser therapy, activates components of the cellular structure, therefore converting luminous energy into photochemical energy and leading to biophysical and biochemical reactions in the mitochondrial respiratory chain. The LLLT promotes cellular and tissue photobiomodulation by means of changes in metabolism, leading to molecular, cellular and systemic changes. The objective of this study was to analyze the action of low-level laser in the hemodynamic modulation of spontaneously hypertensive rats, in the long term. Animals (n = 16) were randomly divided into the Laser Group (n = 8), which received three weekly LLLT irradiations for seven weeks, and into the Sham Group (n = 8), which received three weekly simulations of laser for seven weeks, accounting for 21 applications in each group. After seven weeks, animals were cannulated by the implantation of a catheter in the left carotid artery. On the following day, the systemic arterial pressure was recorded. The Laser Group showed reduced levels of mean blood pressure, with statistically significant reduction (169 ± 4 mmHg* vs. 182 ± 4 mmHg from the Sham Group) and reduced levels of diastolic pressure (143 ± 4 mmHg* vs. 157 ± 3 mmHg from the Sham Group), revealing a 13 and 14 mmHg decrease, respectively. Besides, there was a concomitant important decline in heart rate (312 ± 14 bpm vs. 361 ± 13 bpm from the Sham Group). Therefore, laser therapy was able to produce hemodynamic changes, thus reducing pressure levels in spontaneously hypertensive rats.

Morphological study of adult male worms of Schistosoma mansoni Sambon, 1907 by confocal laser scanning microscopy

Aiming to detail data obtained through brightfield microscopy (BM) on reproductive, excretory and digestive system, specimens of Schistosoma mansoni eight weeks old, were recovered from SW mice, stained with Langeron's carmine and analyzed under a confocal laser scanning microscope CLSM 410 (Carl Zeiss). The reproductive system presented a single and lobate testis, with intercommunications between the lobes without efferent duct. Supernumerary testicular lobe was amorphous and isolated from the normal ones. Collecting tubules (excretory ducts), followed by the excretory bladder, opening to the external media through the excretory pore, were observed at the posterior extremity of the body. In the digestive tract, a cecal swelling was noted at the junction that originates the single cecum. It was concluded that through confocal laser scanning microscopy, new interpretations of morphological structures of S. mansoni worms could be achieved, modifying adopted and current descriptions. The gonad consists of a single lobed testis, similar to that observed in some trematode species. Moreover, the same specimens can be observed either by BM or CLSM, considering that the latter causes only focal and limited damage in tissue structures.

Morphological investigation of Toxoplasma gondii in vivo by a multiple beam interference microscope

A recently developed technique, namely multiple beam interference microscopy, has been applied to investigate the morphology of the parasite Toxoplasma gondii for the first time. The interference pattern obtained from the multiple internal reflection of a T. gondii, sandwiched between a glass plate and a cover plate, was focused on the objective of a conventional microscope. Because of the enhance contrast, several details of sub cellular structure and separating compartments are clearly visible. Details reveal the presence of a nucleus, lipid body, dense granule, rhoptry and amylopectin. The wall thickness of the membrane of the lipid body and the amylopectin is of the order of 0.02 µm and can be clearly distinguished with the help of the present technique. The same parasite has also been examined with the help of atomic force microscopy, and because of its thick membrane, the inner structural details were not observed at all. Sub cellular details of T. gondii observed with the present technique have been reported earlier only by low amplification transmission electron microscopy and not by any optical microscopic technique.