Blue 470 nm light suppresses the growth of Salmonella enterica and methicillin-resistant Staphylococcus aureus (MRSA) in vitro.

BACKGROUND AND OBJECTIVE: Emerging evidence suggests that blue light can photo-inactivate some bacteria of clinical importance. Consequently, we tested the hypothesis that 470 nm light can suppress growth of two recalcitrant bacteria, MRSA and Salmonella. MATERIALS AND METHODS: We plated 5 × 106 and 7 × 106 CFU/ml USA300 strain of MRSA and 1 × 106 CFU/ml of Salmonella enterica serovars Typhimurium and Heidelberg. Plates were irradiated with 55, 110, 165 and 220 J/cm2 of blue light, incubated at 37°C for 24 hours and colony counts determined. RESULTS: Compared with controls, blue light irradiation produced a significant dose-dependent reduction in the number of colonies formed by each bacterial strain (P < 0.001). Irradiation of 5 × 106 and 7 × 106 CFU/ml MRSA with 55 J/cm2 produced 92% (4.6 × 106 CFU/ml) and 86% (6 × 106 CFU/ml) inactivation respectively, while 110 and 220 J/cm2 suppressed each MRSA density 100%. Irradiation of Salmonella Typhimurium with 55 and 110 J/cm2 suppressed bacterial growth 31% (3.1 × 105 CFU/ml) and 93% (9.3 × 105 CFU/ml) respectively; while Salmonella Heidelberg was inhibited 11% (1.1 × 105 CFU/ml) and 84% (8.4 × 105 CFU/ml) respectively by the two fluences. Complete inactivation of each Salmonella strain was achieved using 165 or 220 J/cm2 . CONCLUSION: The observed inhibition of Gram-positive (MRSA) and Gram-negative (Salmonella) bacteria suggests the versatility of blue light in bacteria eradication, making it a viable intervention strategy for decontamination of food and environments that harbor such bacteria. Lasers Surg. Med. 47:595-601, 2015. © 2015 Wiley Periodicals, Inc.

Optimization of the antimicrobial effect of blue light on methicillin-resistant Staphylococcus aureus (MRSA) in vitro.

BACKGROUND AND OBJECTIVE: In previous studies, we showed that irradiation with 405 nm or 470 nm light suppresses up to 92% methicillin-resistant Staphylococcus aureus (MRSA) growth in vitro and that the remaining bacteria re-colonize. In this study, the aim was to develop a protocol that yields 100% MRSA growth suppression. MATERIALS AND METHODS: We cultured 3 × 10(6) and 5 × 10(6) CFU/ml USA300 strain of MRSA and then irradiated each plate with varying fluences of 1-60 J/cm2 of 405 nm or 470 nm light, either once or twice at 6 hours intervals. Next, we plated 7 × 10(6) CFU/ml and irradiated it with 45, 50, 55, or 60 J/cm2 fluence, once, twice, or thrice at the same 6 hours intervals. In a third experiment, the same culture density was irradiated with 0, 165, 180, 220, or 240 J/cm(2) , either once, twice, or thrice. RESULTS: Irradiation with either wavelength significantly reduced the bacterial colonies regardless of bacterial density (P < 0.05). At 3 × 10(6) CFU/ml density, nearly 40% and 50% growth of MRSA were suppressed with as little as 3 J/cm2 of 405 nm and 470 nm wavelengths, respectively. Moreover, 100% of the colonies were suppressed with a single exposure to 55 or 60 J/cm2 of 470 nm light or double treatment with 50, 55, or 60 J/cm2 of 405 nm wavelength. At 5 × 10(6) CFU/ml density, irradiating twice with 50, 55, or 60 J/cm2 of either wavelength suppressed bacterial growth completely, lower fluences did not. The denser 7 × 10(6) CFU/ml culture required higher doses to achieve 100% suppression, either one shot with 220 J/cm2 of 470 nm light or two shots of the same dose using 405 nm. CONCLUSION: The bactericidal effect of blue light can be optimized to yield 100% bacterial growth suppression, but with relatively high fluences for dense bacterial cultures, such as 7 × 10(6) CFU/ml.

The Effects of Blue Light on Human Fibroblasts and Diabetic Wound Healing.

Diabetes is a serious threat to global health and is among the top 10 causes of death. The Diabetic foot ulcer (DFU) is among the most common and severe complications of the disease. Bacterial infections are common; therefore, timely aggressive management, using multidisciplinary management approaches is needed to prevent complications, morbidity, and mortality, particularly in view of the growing cases of antibiotic-resistant bacteria. Photobiomodulation (PBM) involves the application of low-level light at specific wavelengths to induce cellular photochemical and photophysical responses. Red and near-infrared (NIR) wavelengths have been shown to be beneficial, and recent studies indicate that other wavelengths within the visible spectrum could be helpful as well, including blue light (400-500 nm). Reports of the antimicrobial activity and susceptibility of blue light on several strains of the same bacterium show that many bacteria are less likely to develop resistance to blue light treatment, meaning it is a viable alternative to antibiotic therapy. However, not all studies have shown positive results for wound healing and fibroblast proliferation. This paper presents a critical review of the literature concerning the use of PBM, with a focus on blue light, for tissue healing and diabetic ulcer care, identifies the pros and cons of PBM intervention, and recommends the potential role of PBM for diabetic ulcer care.

Pulsed blue light, saliva and curcumin significantly inactivate human coronavirus.

In a recent study, we showed that pulsed blue light (PBL) inactivates as much as 52.3% of human beta coronavirus HCoV-OC43, a surrogate of SARS-CoV-2, and one of the major strains of viruses responsible for the annual epidemic of the common cold. Since curcumin and saliva are similarly antiviral and curcumin acts as blue light photosensitizer, we used Qubit fluorometry and WarmStart RT-LAMP assays to study the effect of combining 405 nm, 410 nm, 425 nm or 450 nm wavelengths of PBL with curcumin, saliva or a combination of curcumin and saliva against human beta coronavirus HCoV-OC43. The results showed that PBL, curcumin and saliva independently and collectively inactivate HCoV-OC43. Without saliva or curcumin supplementation 21.6 J/cm2 PBL reduced HCoV-OC43 RNA concentration a maximum of 32.8% (log10 = 2.13). Saliva supplementation alone inactivated the virus, reducing its RNA concentration by 61% (log10 = 2.23); with irradiation the reduction was as much as 79.1%. Curcumin supplementation alone decreased viral RNA 71.1%, and a maximum of 87.8% with irradiation. The combination of saliva and curcumin reduced viral RNA to 83.1% and decreased the RNA up to 90.2% with irradiation. The reduced levels could not be detected with qPCR. These findings show that PBL in the range of 405 nm to 450 nm wavelength is antiviral against human coronavirus HCoV-OC43, a surrogate of the COVID-19 virus. Further, it shows that with curcumin as a photosensitizer, it is possible to photodynamically inactivate the virus beyond qPCR detectable level using PBL. Since HCoV-OC43 is of the same beta coronavirus family as SARS-CoV-2, has the same genomic size, and is often used as its surrogate, these findings heighten the prospect of similarly inactivating novel coronavirus SARS-CoV-2, the virus responsible for COVID-19 pandemic.

The Antimicrobial Effect of Gold Quantum Dots and Femtosecond Laser Irradiation on the Growth Kinetics of Common Infectious Eye Pathogens: An In Vitro Study.

We studied the antimicrobial effect of gold quantum dots (AuQDs), femtosecond laser irradiation, and the combined effect of laser irradiation and AuQD treatment against common infectious eye pathogens. The INSPIRE HF100 laser system (Spectra Physics) provided a femtosecond laser, which was pumped by a mode-locked femtosecond Ti: sapphire laser MAI TAI HP (Spectra Physics), while a Quanta-Ray nanosecond Nd: YAG laser (Spectra-Physics) was used to precisely synthesize 7.8, 8.7, and 11.6 nm spherical AuQDs. Then, the in vitro growth kinetics and growth rate analysis of E. coli, methicillin-resistant Staphylococcus aureus, Enterococcus faecalis, Listeria monocytogenes, and Candida albicans (treated with the AuQDs, femtosecond laser irradiation, or combined laser and AuQDs treatment) was measured. The biocompatibility of the AuQDs with the retinal epithelial cell lines (ARPE-19) and their toxicity to the cells was assayed. The results showed that (1) in vitro irradiation using a 159 J/cm2 energy density obtained from the 400 nm femtosecond laser suppressed the growth of each of the five pathogens. (2) Similarly, treatment with the AuQDs was antimicrobial against the four bacteria. The AuQDs with an average size of 7.8 nm were more highly antimicrobial and biocompatible and were less cytotoxic than the larger AuQD sizes. (3) The combined femtosecond laser irradiation and AuQD treatment was more highly antimicrobial than each treatment alone. (4) The AuQD treatment did not impair the rate of wound closure in vitro. These findings suggest that combined femtosecond laser irradiation and AuQD treatment is significantly antimicrobial against Candida albicans, Gram-positive L. monocytogenes, S. aureus, and E. faecalis, as well as Gram-negative E. coli. The nontoxicity and biocompatibility of the AuQD particles tested suggest that this form of treatment may be clinically viable.

Pulsed blue light inactivates two strains of human coronavirus.

Emerging evidence suggests that blue light has the potential to inactivate viruses. Therefore, we investigated the effect of 405 nm, 410 nm, 425 nm and 450 nm pulsed blue light (PBL) on human alpha coronavirus HCoV-229 E and human beta coronavirus HCoV-OC43, using Qubit fluorometry and RT-LAMP to quantitate the amount of nucleic acid in irradiated and control samples. Like SARS-CoV-2, HCoV-229E and HCoV-OC43 are single stranded RNA viruses transmitted by air and direct contact; they have similar genomic sizes as SARS-CoV-2, and are used as surrogates for SARS-CoV-2. Irradiation was carried out either at 32.4 J cm-2 using 3 mW cm-2 irradiance or at 130 J cm-2 using 12 mW cm-2 irradiance. Results: (1) At each wavelength tested, PBL was antiviral against both coronaviruses. (2) 405 nm light gave the best result, yielding 52.3% (2.37 log10) inactivation against HCoV-OC43 (p < .0001), and a significant 1.46 log 10 (44%) inactivation of HCoV-229E (p < .01). HCoV-OC43, which like SARS-CoV-2 is a beta coronavirus, was more susceptible to PBL irradiation than alpha coronavirus HCoV-229E. The latter finding suggests that PBL is potentially antiviral against multiple coronavirus strains, and that, while its potency may vary from one virus to another, it seems more antiviral against beta coronaviruses, such as HCoV-OC43. (3) Further, the antiviral effect of PBL was better at a higher irradiance than a lower irradiance, and this indicates that with further refinement, a protocol capable of yielding 100% inactivation of viruses is attainable.

The antimicrobial effect of 400 nm femtosecond laser and silver nanoparticles on gram-positive and gram-negative bacteria.

Silver nanoparticles are well-known for their antimicrobial effect. However, they are potentially toxic in high doses. We explored the possibility of enhancing the bactericidal effect of low concentrations of silver nanoparticles with blue light femtosecond laser irradiation, since such concentrations are less toxic. The growth dynamics of Pseudomonas aeruginosa, Listeria monocytogenes and methicillin-resistant Staphylococcus aureus grown in pre-synthesized silver nanoparticles were measured with or without pre-irradiation with 50 mW and 400 nm femtosecond laser irradiation. With each bacterium, combined treatment with laser and silver nanoparticles significantly reduced bacterial growth, indicating that this form of treatment could be beneficial in the ongoing efforts to reduce the deleterious effects of antibiotic resistant Gram-positive and Gram-negative bacteria. The combined treatment was more antimicrobial than treatment with silver nanoparticles alone or photo-irradiation alone. P. aeruginosa and L. monocytogenes were more susceptible to the bactericidal effects of silver nanoparticles, and the combination of laser treatment and silver nanoparticles than MRSA.

Structural membrane changes induced by pulsed blue light on methicillin-resistant Staphylococcus aureus (MRSA).

BACKGROUND: In a recent study we showed that blue light inactivates methicillin-resistant Staphylococcus aureus (MRSA) by perturbing, depolarizing, and disrupting its cell membrane. PURPOSE: The current study presents visual evidence that the observed biochemical changes also result in cell metabolic changes and structural alteration of the cell membrane. METHODS: Cultures of MRSA were treated with 450 nm pulsed blue light (PBL) at 3 mW/cm2 irradiance, using a sub lethal dose of 2.7 J/cm2 radiant exposure three times at 30-min intervals. Following 24 h incubation at 37 °C, irradiated colonies and control non-irradiated colonies were processed for light and transmission electron microscopy. RESULTS: The images obtained revealed three major effects of PBL; (1) disruption of MRSA cell membrane, (2) alteration of membrane structure, and (3) disruption of cell replication. CONCLUSION: These signs of bacterial inactivation at a dose deliberately selected to be sub-lethal supports our previous finding that rapid depolarization of bacterial cell membrane and disruption of cellular function comprise another mechanism underlying photo-inactivation of bacteria. Further, it affirms the potency of PBL.

Phototherapy promotes healing of chronic diabetic leg ulcers that failed to respond to other therapies.

OBJECTIVE: We tested the hypothesis that combined 660 and 890 nm LED phototherapy will promote healing of diabetic ulcers that failed to respond to other forms of treatment. RESEARCH DESIGN AND METHODS: A double-blind randomized placebo controlled design was used to study 23 diabetic leg ulcers in two groups of 14 patients. Group one ulcers were cleaned, dressed with 1% silver sulfadiazine cream and treated with "placebo" phototherapy (<1.0 J cm(-2)) twice per week, using a Dynatron Solaris 705(R) device. Group two ulcers were treated similarly but received 3 J cm(-2) dose. RESULTS: At each of 15, 30, 45, 60, 75, and 90 days of healing, mean ulcer granulation and healing rates were significantly higher for group two than the "placebo" group (P < 0.02). While "placebo" treated ulcers worsened during the initial 30 days, group two ulcers healed rapidly; achieving 56% more granulation and 79.2% faster healing by day 30, and maintaining similarly higher rates of granulation and healing over the "placebo" group all through. By day 90, 58.3% of group two ulcers had healed fully and 75% had achieved 90-100% healing. In contrast, only one "placebo" treated ulcer healed fully by day 90; no other ulcer attained > or =90% healing. CONCLUSION: Combined 660 and 890 nm light promotes rapid granulation and healing of diabetic ulcers that failed to respond to other forms of treatment.

Visible 405 nm SLD light photo-destroys methicillin-resistant Staphylococcus aureus (MRSA) in vitro.

BACKGROUND: Infections with MRSA remain a growing public health concern, prompting the need to explore alternative treatments instead of the on-going effort to develop stronger drug-based therapies. We studied the effect of 405 nm blue light on two strains of MRSA-US-300 strain of CA-MRSA and the IS853 strain of HA-MRSA-in vitro. METHODS: We cultured and plated each strain, following which bacteria colonies were irradiated with 0, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 25, 30, 35, 40, 45, 50, 55, or 60 J cm(-2) energy densities-just once-using a Solaris superluminous diode (SLD) device. Specimens were incubated at 35 degrees C for 24 hours. Then, digital images obtained were quantified to obtain colony counts and the aggregate area occupied by bacteria colonies. RESULTS: Blue light irradiation produced a statistically significant dose-dependent reduction in both the number and the aggregate area of colonies formed by each bacteria strain (P<0.001). Maximum eradication of the US-300 (92.1%) and the IS-853 colonies (93.5%) was achieved within 9.2 and 8.4 minutes of exposure, respectively. The longer the irradiation the more bacteria were eradicated. However, the effect was non-linear as increases of energy densities between 1.0 and 15 J cm(-2) resulted in more bacteria death than similar increases between 15 and 60 J cm(-2). CONCLUSION: At low doses, blue light photo-destroys HA-MRSA and CA-MRSA in vitro; raising the prospect that phototherapy may be an effective clinical tool in the on-going effort to stem MRSA infections.

Optimal Laser Phototherapy Parameters for Pain Relief.

BACKGROUND AND OBJECTIVE: Studies on laser phototherapy for pain relief have used parameters that vary widely and have reported varying outcomes. The purpose of this study was to determine the optimal parameter ranges of laser phototherapy for pain relief by analyzing data aggregated from existing primary literature. MATERIALS AND METHODS: Original studies were gathered from available sources and were screened to meet the pre-established inclusion criteria. The included articles were then subjected to meta-analysis using Cohen's d statistic for determining treatment effect size. From these studies, ranges of the reported parameters that always resulted into large effect sizes were determined. These optimal ranges were evaluated for their accuracy using leave-one-article-out cross-validation procedure. RESULTS: A total of 96 articles met the inclusion criteria for meta-analysis and yielded 232 effect sizes. The average effect size was highly significant: d = +1.36 [confidence interval (95% CI) = 1.04-1.68]. Among all the parameters, total energy was found to have the greatest effect on pain relief and had the most prominent optimal ranges of 120-162 and 15.36-20.16 J, which always resulted in large effect sizes. The cross-validation accuracy of the optimal ranges for total energy was 68.57% (95% CI = 53.19-83.97). Fewer and less-prominent optimal ranges were obtained for the energy density and duration parameters. None of the remaining parameters was found to be independently related to pain relief outcomes. CONCLUSIONS: The findings of meta-analysis indicate that laser phototherapy is highly effective for pain relief. Based on the analysis of parameters, total energy can be optimized to yield the largest effect on pain relief.

Understanding the antimicrobial activity of selected disinfectants against methicillin-resistant Staphylococcus aureus (MRSA).

Disinfectants and biocidal products have been widely used to combat Methicillin-resistant Staphylococcus aureus (MRSA) infections in homes and healthcare environments. Although disruption of cytoplasmic membrane integrity has been documented as the main bactericidal effect of biocides, little is known about the biochemical alterations induced by these chemical agents. In this study, we used Fourier transform infrared (FT-IR) spectroscopy and chemometric tools as an alternative non-destructive technique to determine the bactericidal effects of commonly used disinfectants against MRSA USA-300. FTIR spectroscopy permits a detailed characterization of bacterial reactivity, allowing an understanding of the fundamental mechanism of action involved in the interaction between bacteria and disinfectants. The disinfectants studied were ethanol 70% (N = 5), isopropanol (N = 5), sodium hypochlorite (N = 5), triclosan (N = 5) and triclocarban (N = 5). Results showed less than 5% colony forming units growth of MRSA treated with triclocarban and no growth in the other groups. Nearly 70,000 mid-infrared spectra from the five treatments and the two control (untreated; N = 4) groups of MRSA (bacteria grown in TSB and incubated at 37°C (Control I) / at ambient temperature (Control II), for 24h) were pre-processed and analyzed using principal component analysis followed by linear discriminant analysis (PCA-LDA). Clustering of strains of MRSA belonging to five treatments and the discrimination between each treatment and two control groups in MRSA (untreated) were investigated. PCA-LDA discriminatory frequencies suggested that ethanol-treated spectra are the most similar to isopropanol-treated spectra biochemically. Also reported here are the biochemical alterations in the structure of proteins, lipid membranes, and phosphate groups of MRSA produced by sodium hypochlorite, triclosan, and triclocarban treatments. These findings provide mechanistic information involved in the interaction between MRSA strains and hygiene products; thereby demonstrating the potential of spectroscopic analysis as an objective, robust, and label-free tool for evaluating the macromolecular changes involved in disinfectant-treated MRSA.

Spectrally resolved infrared microscopy and chemometric tools to reveal the interaction between blue light (470nm) and methicillin-resistant Staphylococcus aureus.

Blue light inactivates methicillin-resistant Staphylococcus aureus (MRSA), a Gram-positive antibiotic resistant bacterium that leads to fatal infections; however, the mechanism of bacterial death remains unclear. In this paper, to uncover the mechanism underlying the bactericidal effect of blue light, a combination of Fourier transform infrared (FTIR) spectroscopy and chemometric tools is employed to detect the photoreactivity of MRSA and its distinctive pathway toward apoptosis after treatment. The mechanism of action of UV light and vancomycin against MRSA is also investigated to support the findings. Principal component analysis followed by linear discriminant analysis (PCA- LDA) is employed to reveal clustering of five groups of MRSA samples, namely untreated (control I), untreated and incubated at ambient air (control II), irradiated with 470nm blue light, irradiated with 253.5 UV light, and vancomycin-treated MRSA. Loadings plot from PCA-LDA analysis reveals important functional groups in proteins (1683, 1656, 1596, 1542cm-1), lipids (1743, 1409cm-1), and nucleic acids region of the spectrum (1060, 1087cm-1) that are responsible for the classification of blue light irradiated spectra and control spectra. Cluster vector plots and scores plot reveals that UV light-irradiated spectra are the most biochemically similar to blue light- irradiated spectra; however, some wavenumbers experience a shift. The shifts between blue light and UV light irradiated loadings plot at νasym PO2- band (from 1228 to 1238cm-1), DNA backbone (from 970 to 966cm-1) and base pairing vibration of DNA (from 1717 to 1712cm-1) suggest distinctive changes in DNA conformation in response to irradiation. Our findings indicate that irradiation of MRSA with 470nm light induces A-DNA cleavage and that B-DNA is more resistant to damage by blue light. Blue light and UV light treatment of MRSA are complementary and distinct from the known antimicrobial effect of vancomycin. Moreover, it is known that UV-induced cleavage of DNA predominantly targets B-DNA, which is in agreement with the FTIR findings. Overall the results suggest that the combination of light and vancomycin could be a more robust approach in treating MRSA infections.

Blue/violet laser inactivates methicillin-resistant Staphylococcus aureus by altering its transmembrane potential.

The resistance of methicillin-resistant Staphylococcus aureus to antibiotics presents serious clinical problems that prompted the need for finding alternative or combination therapies. One such therapy is irradiation with blue light. To determine the alterations in metabolic processes implicated in the observed antimicrobial effects of blue light, we investigated the changes in membrane potential and the presence of free-radical-producing photo-acceptor molecules. Bacterial cultures irradiated with one or two doses of 405nm laser light (each consisting of 121J/cm2) were imaged with spectrally resolved laser-scanning microscopes to detect endogenous fluorescent species as well as the voltage sensitive dye 3,3'-Diethyloxacarbocyanine iodide. The endogenous fluorescence indicated the presence of photosensitizers (i.e., porphyrins, NADH, FAD) in the cells, while the exogenous signal allowed us to monitor rapid changes in transmembrane potential following treatment with light. The changes were drastic within the first 5min after irradiation with the first dose and continued slowly after the second irradiation. These results suggest that the early antimicrobial activity of blue light results from alteration of membrane integrity with a consequent decrease in membrane polarization and rapid alteration of vital cellular functions. The observation of an early antimicrobial activity of light is very encouraging, as it suggests that treatment does not necessarily have to be administered over a long period of time.

A comparison of four methods for determining viability in human dermal fibroblasts irradiated with blue light.

INTRODUCTION: Several tests are available for assessing the viability of cells; however, there is a dearth of studies comparing the results obtained with each test. We compared the capability of four viability assays (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (MTT), neutral red, trypan blue and live/dead fluorescence), to detect potential toxicity in fibroblasts irradiated with 470nm blue light. METHODS: Cells were irradiated at 3, 55, 110 and 220J/cm(2), incubated for 24h and viability assessed using each test. RESULTS: MTT assay showed significant decreases in viability when cells were irradiated with 110 and 220J/cm(2) energy fluence (dose) (89% and 57% viable cells, respectively; p<0.0001, compared to control); likewise the trypan blue assay showed 42% and 46% viable cells (p<0.0001). Neutral red assay revealed significant decrease in viability when cells were irradiated with 220J/cm(2) (84% viable cells; p=0.0008, compared to control). The live/dead fluorescence assay was less sensitive, evincing 91% and 95% viable cells after irradiation with 110 and 220J/cm(2) respectively. DISCUSSION: (1) The four assays differed in their levels of sensitivity to cell viability. (2) The adverse effect of increasing doses seems to manifest as alteration of mitochondrial metabolism, followed by lysosomal dysfunction, membrane disruption and finally loss of cell membrane integrity. (3) Overall, irradiation with 3J/cm(2) or 55J/cm(2) did not adversely affect cell viability. Thus, doses below 110J/cm(2) appear safe.

Whole-Genome Sequence for Methicillin-Resistant Staphylococcus aureus Strain ATCC BAA-1680.

We report here the whole-genome sequence of the USA300 strain of methicillin-resistant Staphylococcus aureus (MRSA), designated ATCC BAA-1680, and commonly referred to as community-associated MRSA (CA-MRSA). This clinical MRSA isolate is commercially available from the American Type Culture Collection (ATCC) and is widely utilized as a control strain for research applications and clinical diagnosis. The isolate was propagated in ATCC medium 18, tryptic soy agar, and has been utilized as a model S. aureus strain in several studies, including MRSA genetic analysis after irradiation with 470-nm blue light.

Soft tissue thermodynamics before, during, and after cold pack therapy.

BACKGROUND: Cold packs are commonly used by clinicians, trainers, and others, often as an interim treatment for many acute conditions, but the extent of temperature change associated with this form of treatment remains poorly understood. METHODS: In 16 healthy male and female volunteers aged 25.4 +/- 3.6 yr, we monitored skin temperature, and recorded the temperature of the quadriceps muscle at 1, 2, and 3 cm depths below the skin, before, during, and after 20 min of cold pack treatment. RESULTS: The results revealed a slight rise in temperature at all four levels during the 5 min pretreatment period, but significant temperature falls at the skin and 1 cm levels beginning from 8 min of treatment (P < 0.001). There was no significant change in tissue temperature at the 2.0 cm or 3.0 cm depths throughout treatment. However, after treatment, cutaneous temperature and the temperature at 1.0 cm depth rose rapidly, returning to baseline levels at variable intersubject times. As these superficial temperatures rose, there were concurrent falls in the temperatures at the 2.0 cm and 3.0 cm levels. Thus, the deeper tissues lost heat (cooled) simultaneously as the superficial tissues rewarmed; to the extent that 40 min after treatment, the deeper levels were cooler than the cutaneous and 1.0 cm levels. CONCLUSION: 1) Cold pack therapy produces significant temperature falls in cutaneous and subcutaneous superficial tissues without directly changing the temperature of tissues at or more than 2.0 cm below the skin; and 2) the temperature gradients of both layers of tissue reverses after treatment, indicating that the deep tissue beneath is at least one of the sources of heat used to rewarm the cooled superficial tissue. The latter finding underscores the importance of the hemodynamic interchange between superficial and deep tissues, and offers an explanation for the reduction of pain, muscle spasm, and edema observed with cold therapy in several clinical situations.

An inexpensive, automated instrument for laser irradiation of cultured cells.

OBJECTIVE: Laser irradiation of cultured cells is a valuable technique for elucidating the mechanisms of low-level laser therapy, but is often tedious because of the need to manually change the position of the laser beam. Consequently, we developed a computer-based system that automatically moves a cell culture plate over a laser beam and times the exposure. BACKGROUND DATA: There are presently no commercial devices available for automated laser irradiation of cultured cells. Many investigators thus manually aim and time laser exposure, a time-consuming task that is prone to errors. MATERIALS AND METHODS: We used outdated, surplus computer components to construct a system for automated laser exposure of cultured cells. This design strategy makes the system quite inexpensive. RESULTS: Construction and operation of the system is described and an example of its use is presented. Alternate means of accomplishing automated laser irradiation are also presented. CONCLUSION: Inexpensive and relatively simple devices can be constructed for automated laser irradiation of cultured cells. These devices can eliminate the tedium and errors of manual laser exposure.

The efficacy of low-power lasers in tissue repair and pain control: a meta-analysis study.

OBJECTIVE: We used statistical meta-analysis to determine the overall treatment effects of laser phototherapy on tissue repair and pain relief. BACKGROUND DATA: Low-power laser devices were first used as a form of therapy more than 30 years ago. However, their efficacy in reducing pain or promoting tissue repair remains questionable. METHODS: Following a literature search, studies meeting our inclusion criteria were identified and coded. Then, the effect size of laser treatment, that is, Cohen's d, was calculated from each study using standard meta-analysis procedures. RESULTS: Thirty-four peer-reviewed papers on tissue repair met our inclusion criteria and were used to calculate 46 treatment effect sizes. Nine peer-reviewed papers on pain control met the inclusion criteria and were used to calculate nine effect sizes. Meta-analysis revealed a positive effect of laser phototherapy on tissue repair (d = +1.81; n = 46) and pain control (d = +1.11; n = 9). The positive effect of treatment on specific indices of tissue repair was evident in the treatment effect sizes determined as follows: collagen formation (d = +2.78), rate of healing (d = +1.57), tensile strength (d = +2.13), time needed for wound closure (d = +0.76), tensile stress (d = +2.65), number and rate of degranulation of mast cells (d = +1.87), and flap survival (d = +1.95). Further, analysis revealed the positive effects of various wavelengths of laser light on tissue repair, with 632.8 nm having the highest treatment effect (d = +2.44) and 780 nm the least (d = 0.60). The overall treatment effect for pain control was positive as well (d = +1.11). The fail-safe number-that is, the number of studies in which laser phototherapy has negative or no effect-needed to nullify the overall outcome of this analysis was 370 for tissue repair and 41 for pain control. CONCLUSIONS: These findings mandate the conclusion that laser phototherapy is a highly effective therapeutic armamentarium for tissue repair and pain relief.