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Four-hour processing of clinical/diagnostic specimens for electron microscopy using microwave technique.

A protocol for routine 4-hour microwave tissue processing of clinical or other samples for electron microscopy was developed. Specimens are processed by using a temperature-restrictive probe that can be set to automatically cycle the magnetron to maintain any designated temperature restriction (temperature maximum). In addition, specimen processing during fixation is performed in 1.7-ml microcentrifuge tubes followed by subsequent processing in flow-through baskets. Quality control is made possible during each step through the addition of an RS232 port to the microwave, allowing direct connection of the microwave oven to any personal computer. The software provided with the temperature probe enables the user to monitor time and temperature on a real-time basis. Tissue specimens, goat placenta, mouse liver, mouse kidney, and deer esophagus were processed by conventional and microwave techniques in this study. In all instances, the results for the microwave-processed samples were equal to or better than those achieved by routine processing techniques.

Adenoviridae Infections↗

Effect of fixation time and microwave oven heating time on retrieval of the Ki-67 antigen from paraffin-embedded tissue.

Microwave oven heating has been employed for retrieval of antigens from formalin-fixed, paraffin-embedded tissue for immunohistochemical staining. Recently, a Ki-67 equivalent murine monoclonal antibody was generated which can detect tumor proliferative activity in routinely processed tissue with microwave oven heating. We assessed the effect of fixation time (4, 24, or 48 hr) and microwave oven heating time (7, 14, 21, 28, 35, or 49 min) on retrieval of the Ki-67 antigen from tonsil tissue. Ki-67 staining was quantitated by image analysis. Owing to the heterogeneity of Ki-67 staining within and between germinal centers, we employed a measurement technique that averages staining across the germinal centers of each section. Good Ki-67 immunostaining was observed for all microwave oven heating times in tissue fixed for 4 hr. In contrast, poor immunostaining was observed in tissue fixed for 48 hr unless a heating time of at least 14 min was used. Tonsil fixed for 24 or 48 hr showed a significant increase in percentage positive nuclear area after microwave times of 14 or 21 min. Prolonged heating time (up to 49 min) had no effect on the quality of Ki-67 staining in tissue fixed for 4 or 48 hr.

Formaldehyde↗

[Effects of whole-body microwave exposure on the plasma adrenocorticotropic hormone, thyroid-stimulating hormone and thyroid hormones in rats].

To investigate the effects of whole-body microwave exposure on plasma adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), thyroxine (T4) and free triiodothyronine (T3) levels, rats weighing 267-306 g (light group) and 352-387 g (heavy group) were exposed to microwaves with a frequency of 2,450 MHz at the power densities of 5 and 10 mW/cm2 under an ambient temperature of 21 degrees C to 23 degrees C for an hour. Rectal temperatures were increased by 1.0 degrees C at 5 mW/cm2 and 2.1 degrees C at 10 mW/cm2 in the light group, and by 0.7 degrees C at 5 mW/cm2 and 1.5 degrees C at 10 mW/cm2 in the heavy group after microwave exposure. Plasma levels of ACTH increased significantly at 10 mW/cm2 in the light group and 5 and 10 mW/cm2 in the heavy group after microwave exposure. Plasma levels of TSH increased significantly at 10 mW/cm2 only in the light group. As for plasma levels of free T3, a significant decrease at 10 mW/cm2 was observed only in the heavy group. However, plasma levels of T4 did not change in either group after microwave exposure.

Adrenocorticotropic Hormone↗

Effects of hyperthermia induced by microwave irradiation on brain development in mice.

Pregnant mice were exposed to 2.45 GHz of microwave radiation for 15 or 20 min on day 13 of gestation. The highest maternal core temperature during the exposure did not exceed 42.5 degrees C. Pregnant females also were immersed in hot water at 42 degrees C for 15 min to compare thermal effects on brain development. Animals were killed 9 hours after treatment, and the pyknotic cells in the ventricular zone of telencephalon were counted. The respective incidences of these cells in the groups exposed to microwaves for 15 and 20 min were 1.83% and 3.06%. Microwave radiation for 20 min had an effect that was comparable to that of immersion in 42 degrees C hot water for 15 min. In addition, some animals were examined on day 18 of gestation, and some of their offspring were examined at 6 weeks of age in an examination of long-term effects. Brain weight for the group exposed to microwaves for 20 min was significantly lower than for the control group, and the numerical density of the neurons in the cerebrum was higher. We concluded that microwave radiation at the dose tested mainly has a thermal effect.

Animals↗

Effect of microwave treatments on dimensional accuracy of maxillary acrylic resin denture base.

Microwave energy has been used as an alternative method for disinfection and sterilization of dental prostheses. This study evaluated the influence of microwave treatment on dimensional accuracy along the posterior palatal border of maxillary acrylic resin denture bases processed by water-bath curing. Thirty maxillary acrylic bases (3-mm-thick) were made on cast models with Clássico acrylic resin using routine technique. After polymerization and cooling, the sets were deflasked and the bases were stored in water for 30 days. Thereafter, the specimens were assigned to 3 groups (n=10), as follows: group I (control) was not submitted to any disinfection cycle; group II was submitted to microwave disinfection for 3 min at 500 W; and in group III microwaving was done for 10 min at 604 W. The acrylic bases were fixed on their respective casts with instant adhesive (Super Bonder) and the base/cast sets were sectioned transversally in the posterior palatal zone. The existence of gaps between the casts and acrylic bases was assessed using a profile projector at 5 points. No statistically significant differences were observed between the control group and group II. However, group III differed statistically from the others (p<0.05). Treatment in microwave oven at 604 W for 10 min produced the greatest discrepancies in the adaptation of maxillary acrylic resin denture bases to the stone casts.

Acrylic Resins↗

Joint effects of microwave and chromium trioxide on root tip cells of Vicia faba.

The mutagenic effects of microwave and chromium trioxide (CrO(3)) on Vicia faba root tip were studied. Micronucleus assay and chromosomal aberration assay were used to determine the mitotic index, the micronucleus frequency and chromosomal aberration frequency of Vicia faba root tip cells induced by microwave and CrO(3). The results showed that the micronucleus frequency decreased, and that the mitotic index and chromosomal aberration frequency showed linear dose responses to CrO(3), in treatment of microwave for 5 s. In microwave of 25 s, the mitotic index decreased, the micronucleus frequency and chromosomal aberration frequency increased with increase of CrO(3) concentration. We concluded that microwave and CrO(3) had antagonistic effect on the mitotic index of Vicia faba root tip cells, but had synergetic effect on micronucleus frequency and chromosomal aberration frequency of Vicia faba root tip cells.

Cells, Cultured↗

Exposure of cultured astroglial and microglial brain cells to 900 MHz microwave radiation.

The rapid rise in the use of mobile communications has raised concerns about health issues related to low-level microwave radiation. The head and brain are usually the most exposed targets in mobile phone users. In the brain, two types of glial cells, the astroglial and the microglial cells, are interesting in the context of biological effects from microwave exposure. These cells are widely distributed in the brain and are directly involved in the response to brain damage as well as in the development of brain cancer. The aim of the present study was to investigate whether 900 MHz radiation could affect these two different glial cell types in culture by studying markers for damage-related processes in the cells. Primary cultures enriched in astroglial cells were exposed to 900 MHz microwave radiation in a temperature-controlled exposure system at specific absorption rates (SARs) of 3 W/kg GSM modulated wave (mw) for 4, 8 and 24 h or 27 W/kg continuous wave (cw) for 24 h, and the release into the extracellular medium of the two pro-inflammatory cytokines interleukin 6 (Il6) and tumor necrosis factor-alpha (Tnfa) was analyzed. In addition, levels of the astroglial cell-specific reactive marker glial fibrillary acidic protein (Gfap), whose expression dynamics is different from that of cytokines, were measured in astroglial cultures and in astroglial cell-conditioned cell culture medium at SARs of 27 and 54 W/kg (cw) for 4 or 24 h. No significant differences could be detected for any of the parameters studied at any time and for any of the radiation characteristics. Total protein levels remained constant during the experiments. Microglial cell cultures were exposed to 900 MHz radiation at an SAR of 3 W/kg (mw) for 8 h, and I16, Tnfa, total protein and the microglial reactivity marker ED-1 (a macrophage activation antigen) were measured. No significant differences were found. The morphology of the cultured astroglial cells and microglia was studied and appeared to be unaffected by microwave irradiation. Thus this study does not provide evidence for any effect of the microwave radiation used on damage-related factors in glial cells in culture.

Animals↗

The application of microwave irradiation as new convenient synthetic procedure in drug discovery.

Heterocyclic compounds hold a special place among pharmaceutically important natural and synthetic materials. The remarkable ability of heterocyclic nuclei to serve both as biomimetics and reactive pharmacophores has largely contributed to their unique value as traditional key elements of numerous drugs. In both lead identification and lead optimization processes there is an acute need for new organic small molecules. Traditional methods of organic synthesis are orders of magnitude too slow to satisfy the demand for these compounds. The fields of combinatorial and automated medicinal chemistry have been developed to meet the increasing requirement of new compounds for drug discovery, within these fields, speed is of the essence. The efficiency of microwave flash-heating chemistry in dramatically reducing reaction times (reduced from days and hours to minutes and seconds) has recently been proven in several different fields of organic chemistry. We believe that the time saved by using focused microwaves is potentially important in traditional organic synthesis but could be of even greater importance in high-speed combinatorial and medicinal chemistry. In this review, it is impossible to cover all significant developments in the area of microwave-assisted organic synthesis (MAOS). Rather, outlines the basic principles behind the technology and summarizes the areas in which microwave technology has made an impact, to date. Specific attention is given to application of microwave irradiation in liquid systems, and in the solid state as well of several representative biologically interesting nuclei. In addition we report some of the most recently disclosed applications in combinatorial chemistry.

Combinatorial Chemistry Techniques↗

Sonographically guided microwave coagulation treatment of liver cancer: an experimental and clinical study.

OBJECTIVE: Percutaneous microwave coagulation was performed with a modified system in animal experiments and in a clinical study to evaluate this technique as a treatment option for liver cancer. SUBJECTS AND METHODS: As an in vitro study, a microwave electrode was inserted 5-6 cm into separated egg white, homogenate of pig liver, and pig liver, with different power outputs and different lengths of inner conductors. In the animal experiment, the sonographically guided coagulation was performed percutaneously nine times and at laparotomy 43 times on 17 adult dogs. The thermal needles were placed parallel to and 5 mm, 8-12 mm, and 15 mm from the electrode. Clinically, 41 patients with hepatocellular carcinoma and 10 patients with hepatic metastases were treated with a 60-W microwave emission for 240-300 sec. RESULTS: Microwave coagulation using the modified system at 60 W for 300 sec produced a necrosis volume of 3.7 x 2.6 x 2.6 cm. The coagulated volume was elliptic when the exposed inner conductor of the electrode was 27 mm. The temperature at the periphery was 62.0 +/- 5.8 degrees C. During a mean follow-up period of 23 months, in 41 patients with hepatocellular carcinoma, 79% (46/58) of lesions became smaller, and the intratumoral blood flow disappeared in 89% (47/53). All tumors showed decreased density on unenhanced CT, and 84% (32/38) of tumors showed no enhancement on contrast-enhanced CT. In 21 patients with an elevated alpha-fetoprotein level, the level decreased in all 21 and was normalized in 17. A second biopsy on 19 patients showed complete destruction of tumor in 18. In 10 patients with hepatic metastases, the mean follow-up period was 13 months. Shrinkage of lesions occurred in 84% (21/25), and the blood flow inside the tumor disappeared in 75% (12/16) of lesions. Seventy-three percent (8/11) of the nodules showed no enhancement. A second biopsy on six patients showed complete necrosis in five. CONCLUSION: Sonographically guided microwave coagulation performed with this modified system was an effective and safe treatment for liver cancer.

Adult↗

Sonography-guided percutaneous microwave ablation of high-grade dysplastic nodules in cirrhotic liver.

OBJECTIVE: Our objective was to evaluate the effect of sonography-guided percutaneous microwave ablation of high-grade dysplastic nodules in the cirrhotic liver. MATERIALS AND METHODS: From July 1997 to May 2003, 49 histologically proven high-grade dysplastic nodules in 30 patients with liver cirrhosis were treated by microwave ablation. Three patients had concomitant small hepatocellular carcinomas (D < 3.0 cm), whereas another three had undergone liver segmentectomy for hepatocellular carcinoma 1 year earlier. The mean size of the nodules was 1.8 cm (range, 0.9-4.6 cm). Sixty-eight insertions with 78 applications were administered to the 49 nodules. RESULTS: The follow-up period was 12-82 months (mean, 45.1 +/- 19.0 months). Five patients died during this study: three from advanced hepatocellular carcinoma, one from bleeding in the upper gastrointestinal tract, and another from cerebral hemorrhage. All nodules showed decreased density on unenhanced CT and no enhancement on contrast-enhanced CT after microwave ablation. Posttreatment biopsy performed in 16 patients with 18 nodules 1-3 months after microwave ablation showed no evidence of viable tissue but replacement by fibrotic tissue in all nodules. CONCLUSION: Percutaneous microwave ablation as a minimally invasive therapy is effective for ablating high-grade dysplastic nodules, thus preventing their potential malignant transformation, which may improve survival. The preliminary data warrant further prospective, randomized studies.

Adult↗

Magnetic resonance-guided percutaneous microwave coagulation therapy for liver metastases of breast cancer in a case.

Real-time magnetic resonance (MR) imaging enables the application of percutaneous microwave coagulation for high-risk patients with metastatic liver tumours. The tumours, local vessels and bile ducts can be observed clearly in three-dimensional sections and a sufficient surgical margin can be confirmed on the MR image even during the coagulation procedure. MR-guided percutaneous microwave coagulation therapy is effective for treatment of not only primary liver tumours but also metastatic breast cancers in the liver, which are not diffuse but discrete, and difficult to treat with only chemo-and endocrine therapy. We report a 44-year-old Japanese woman who underwent modified radical mastectomy for right breast cancer (T1c N0 M0 Stage I). Three years after the operation, she developed two metastatic liver tumours and was treated by MR-guided percutaneous microwave coagulation, achieving a complete response (CR) without any recurrence for 15 months as of the present. The most beneficial aspect of MR-guided percutaneous microwave coagulation is its safety. It is only minimally invasive and can be repeated. This therapy, therefore promises to prolong the disease free period. Additional clinical trials will be valuable to delineate the effectiveness and safety of MR-guided percutaneous microwave coagulation therapy for controlling the liver metastases of breast cancer.

Adult↗

Microwave fixation in diagnostic renal pathology.

Microwave-fixed tissues were examined in 10 patients undergoing diagnostic renal biopsy. A small portion of renal tissue was fixed by microwave irradiation and subsequently processed by routine methods for light microscopic, immunofluorescent and electron microscopic studies. The remaining portion of specimen was fixed and processed by conventional methods. In light microscopic examination, the renal architecture and cell morphology were well-preserved. Pathological changes were identical to those seen with formalin-fixed tissue. The pattern, distribution and intensity of positive immunofluorescence in microwave-fixed tissue were similar to those in tissues directly snap-frozen and stained. In electron microscopy, the normal and pathological features were well-demonstrated and not different from those observed in glutaraldehyde-fixed specimens. Specific ultrastructural lesions were clearly demonstrated and, apparently, were not altered by microwave irradiation. Our preliminary data indicate that microwave fixation can be effectively applied in the processing of renal biopsies. As the fixation is rapid, this method may be valuable in circumstances when an urgent diagnosis is required.

Biopsy↗

Clinical research into hyperthermia treatment of cancer using a 430 MHz microwave heating system with a lens applicator.

Tokyo Metropolitan Komagome Hospital, Kyoto University Hospital, and Aichi Cancer Center used a lens applicator microwave equipment, the HTS-100, to treat 90 patients with a total of 96 tumours. The results were analysed, and the following conclusions reached. Three clinical cases have been presented to illustrate the features of the HTS-100. (1) The results of 383 heating sessions using the HTS-100 were analysed. Even tumours larger and deeper than 5 cm could be heated above 42 degrees C. The heating area is much wider than that of the other 2450, 915, 430 MHz microwave heating equipments. (2) Hyperthermia treatment using the HTS-100 was combined with radiotherapy. The percentage effectiveness in terms of CR+PRa was 66.3% for the 89 cases which could be evaluated. Even for tumours larger than 5 cm, and deeper than 4 cm, local control was achieved in 60.7%. Thus, the indications for microwave heating have been significantly expanded. (3) Combination of microwave heating and RF heating is a new protocol which also expands the indications for hyperthermia therapy. (4) The percentage of patients experiencing the side-effects of pain, a sensation of heat, and burns was 15.6%, 6.3%, and 3.6%, respectively. Most of these side-effects were mild and temporary. The percentage of patients experiencing pain was higher than in conventional microwave heating, probably because the area of heating was wider. (5) In the future, the HTS-100 can be expected to play an important role in clinical cancer hyperthermia.

Adolescent↗

Thermoregulation in the canine prostate during transurethral microwave hyperthermia, Part I: Temperature response.

In this research, experiments were performed to study thermoregulation in the canine prostate during microwave hyperthermia. The transurethral thermal therapy (T3) system provided by Urologix, Inc. was used to impose microwave heating in the canine prostate. Five types of temperature responses to different microwave power levels as time varies, including both damped and sustained oscillatory temperature responses, have been observed. Decreases in prostatic tissue temperature during the microwave heating are believed to be caused by the increase in local blood flow stimulated by tissue temperature elevations. In this study, the characteristic temperatures and time associated with each response. This work will help to provide a better understanding of how tissue temperature is regulated within the canine prostate during transurethral microwave hyperthermia. Results of the present study offer an experimental foundation for a more detailed theoretical analysis on the temperature distribution based on the power input and local blood perfusion.

Animals↗

Effects of microwaves on the colony-forming capacity of haemopoietic stem cells in mice.

A suspension of bone marrow cells from femurs of female (CBA X C57Bl)F1 mice was exposed to 2450 MHz CW microwaves in a specially designed waveguide exposure system. The temperature of the suspension rose, during exposure to microwaves, from 20 degrees C to 45 degrees C, and at an interval within 20 degrees C to 45 degrees C the number of haemopoietic stem cells (CFUs) was determined by the spleen exocolony method. The time of exposure of bone marrow cells to each temperature studied was 20 s. Control suspensions of bone marrow cells were exposed to a water bath temperature. There were no significant effects of the CFUs with the water bath temperature, while after exposure to microwaves the number of spleen colonies was elevated with a nadir at the temperature of 37 degrees C. With a microwave-induced increase of the temperature above 41 degrees C the number of CFUs in the bone marrow suspension decreased. The increase in the number of colonies was related to the rise in the seeding rate of the CFUs as well as to a rise in their proliferative activity, while the drop in the number of colonies was influenced also by heat-killing of the CFUs by microwave exposure.

Animals↗

The effect of microwave heating on vitamins B1 and E, and linoleic and linolenic acids, and immunoglobulins in human milk.

Breast milk was treated with (1) conventional heating (in water bath) vs microwave heating; (2) microwave heating at two power levels (30% and 100%); (3) increasing final temperatures; and (4) microwave thawing vs refrigerator thawing and examined for changes in specific immunoglobulins to a pool of E. coli and poliovirus type 1 antigens, vitamins E and B1, and the polyunsaturated fatty acids linoleic and linolenic acid. Immunoglobulin activities were stable until final milk temperatures of around 60-65 degrees C were reached, and total inactivation occurred at 77 degrees C. Heating even to high final temperatures did not change contents of vitamins and polyunsaturated fatty acids. No differences in immunoglobulins and nutrients were demonstrated between microwave heating and conventional heating, and between power levels or thawing methods. The study shows that microwave heating of human milk can be performed without significant losses of examined immunoglobulins and nutrients, provided that final temperatures are below 60 degrees C.

Antibodies, Bacterial↗

Influence of gamma-irradiation and microwaves on the antioxidant property of some essential oils.

The antioxidant property of anise, caraway, cumin and fennel essential oils extracted from untreated, gamma-irradiated and microwaved fruits against sunflower oil oxidative rancidity was evaluated. The fruits were exposed to gamma-irradiation at 10 KGy and to microwaves at low oven power setting for 1 min. The essential oils were added individually (200 ppm) to sunflower oil and the rate of oil oxidation was followed by determining the peroxide value during storage at room temperature. The irradiated and microwaved essential oils exhibited an antioxidant activity and was superior to that of sunflower oil catalysed by a mixture of BHT + BHA (200 ppm) in most cases. The present data show that gamma-irradiation and microwave treatments did not affect the antioxidant property of the essential oils under study. In addition the essential oils extracted from the gamma-irradiated fruits were more effective as an antioxidant in sunflower oil than those produced from microwaved fruits.

Analysis of Variance↗

Safranin O staining using a microwave oven.

We investigated the effects of microwave irradiation on a safranin O staining method for paraffin sections of formalin fixed rabbit larynx. The control sections were stained according to the conventional method, and the experimental sections were stained in microwave oven for 10 sec at 360 W in Weigert's iron hematoxylin, and for 30 sec at 360 W in fast green and 0.1% safranin O staining solutions. Light microscopic examination of the sections revealed that the microwave heating did not adversely affect the staining properties of cartilage tissue compared to the conventional staining method. Small differences such as darker staining of the matrix and shrinkage of the cytoplasm was observed in some microwave treated sections. The present study revealed that microwave application can be used safely for the safranin O method with the advantage of reduced staining time.

Animals↗