Applicability of Gene Expression in Saliva as an Alternative to Blood for Biodosimetry and Prediction of Radiation-induced Health Effects.

As the great majority of gene expression (GE) biodosimetry studies have been performed using blood as the preferred source of tissue, searching for simple and less-invasive sampling methods is important when considering biodosimetry approaches. Knowing that whole saliva contains an ultrafiltrate of blood and white blood cells, it is expected that the findings in blood can also be found in saliva. This human in vivo study aims to examine radiation-induced GE changes in saliva for biodosimetry purposes and to predict radiation-induced disease, which is yet poorly characterized. Furthermore, we examined whether transcriptional biomarkers in blood can also be found equivalently in saliva. Saliva and blood samples were collected in parallel from radiotherapy (RT) treated patients who suffered from head and neck cancer (n = 8) undergoing fractioned partial-body irradiations (1.8 Gy/fraction and 50-70 Gy total dose). Samples were taken 12-24 h before first irradiation and ideally 24 and 48 h, as well as 5 weeks after radiotherapy onset. Due to the low quality and quantity of isolated RNA samples from one patient, they had to be excluded from further analysis, leaving a total of 24 saliva and 24 blood samples from 7 patients eligible for analysis. Using qRT-PCR, 18S rRNA and 16S rRNA (the ratio being a surrogate for the relative human RNA/bacterial burden), four housekeeping genes and nine mRNAs previously identified as radiation responsive in blood-based studies were detected. Significant GE associations with absorbed dose were found for five genes and after the 2nd radiotherapy fraction, shown by, e.g., the increase of CDKN1A (2.0 fold, P = 0.017) and FDXR (1.9 fold increased, P = 0.002). After the 25th radiotherapy fraction, however, all four genes (FDXR, DDB2, POU2AF1, WNT3) predicting ARS (acute radiation syndrome) severity, as well as further genes (including CCNG1 [median-fold change (FC) = 0.3, P = 0.013], and GADD45A (median-FC = 0.3, P = 0.031)) appeared significantly downregulated (FC = 0.3, P = 0.01-0.03). A significant association of CCNG1, POU2AF1, HPRT1, and WNT3 (P = 0.006-0.04) with acute or late radiotoxicity could be shown before the onset of these clinical outcomes. In an established set of four genes predicting acute health effects in blood, the response in saliva samples was similar to the expected up- (FDXR, DDB2) or downregulation (POU2AF1, WNT3) in blood for up to 71% of the measurements. Comparing GE responses (PHPT1, CCNG1, CDKN1A, GADD45A, SESN1) in saliva and blood samples, there was a significant linear association between saliva and blood response of CDKN1A (R2 = 0.60, P = 0.0004). However, the GE pattern of other genes differed between saliva and blood. In summary, the current human in vivo study, (I) reveals significant radiation-induced GE associations of five transcriptional biomarkers in salivary samples, (II) suggests genes predicting diverse clinical outcomes such as acute and late radiotoxicity as well as ARS severity, and (III) supports the view that blood-based GE response can be reflected in saliva samples, indicating that saliva is a "mirror of the body" for certain but not all genes and, thus, studies for each gene of interest in blood are required for saliva.

Development of a Point-of-Care Microfluidic RNA Extraction Slide for Gene Expression Diagnosis after Irradiation.

In times of war, radiological/nuclear emergency scenarios have become a reemphasized threat. However, there are challenges in transferring whole-blood samples to laboratories for specialized diagnostics using RNA. This project aims to miniaturize the process of unwieldy conventional RNA extraction with its stationed technical equipment using a microfluidic-based slide (MBS) for point-of-care diagnostics. The MBS is thought to be a preliminary step toward the development of a so-called lab-on-a-chip microfluidic device. A MBS would enable early and fast field care combined with gene expression (GE) analysis for the prediction of hematologic acute radiation syndrome (HARS) severity or identification of RNA microbes. Whole blood samples from ten healthy donors were irradiated with 0, 0.5 and 4 Gy, simulating different ARS severity degrees. RNA quality and quantity of a preliminary MBS was compared with a conventional column-based (CB) RNA extraction method. GE of four HARS severity-predicting radiation-induced genes (FDXR, DDB2, POU2AF1 and WNT3) was examined employing qRT-PCR. Compared to the CB method, twice as much total RNA from whole blood could be extracted using the MBS (6.6 ± 3.2 µg vs. 12.0 ± 5.8 µg) in half of the extraction time, and all MBS RNA extracts appeared DNA-free in contrast to the CB method (30% were contaminated with DNA). Using MBS, RNA quality [RNA integrity number equivalent (RINe)] values decreased about threefold (3.3 ± 0.8 vs. 9.0 ± 0.4), indicating severe RNA degradation, while expected high-quality RINe ≥ 8 were found using column-based method. However, normalized cycle threshold (Ct) values, as well as radiation-induced GE fold-changes appeared comparable for all genes utilizing both methods, indicating that no RNA degradation took place. In summary, the preliminary MBS showed promising features such as: 1. halving the RNA extraction time without the burden of heavy technical equipment (e.g., a centrifuge); 2. absence of DNA contamination in contrast to CB RNA extraction; 3. reduction in blood required, because of twice the biological output of RNA; and 4. equal GE performance compared to CB, thus, increasing its appeal for later semi-automatic parallel field applications.

The efficacy and the adverse reactions of laser-assisted tattoo removal - a prospective split study using nanosecond and picosecond lasers.

BACKGROUND: Laser pulses with nanosecond duration (NSL) have been the golden standard to destroy the pigment particles in skin. It is still controversially discussed whether picosecond pulses (PSL) are superior for tattoo removal. OBJECTIVES: To compare the efficacy and the adverse reactions of nanosecond and picosecond laser pulses in a comparative study. METHODS: The prospective study included 23 subjects with 30 black or coloured tattoos, which were split into two halves treated with either a new PSL (532, 1064 nm) or standard NSL (694 nm). The lasers were applied at regular time intervals of 4 weeks for up to eight treatments. Tattoo clearance (primary endpoint), pain and adverse reactions (secondary endpoints) were appraised by physicians, blinded observers, and by subjects. The extent and duration of adverse reactions were additionally assessed by using a questionnaire and photo-documentation after each treatment session. RESULTS: The tattoo clearance appeared to be more effective for PSL compared to NSL but without statistical significance (P > 0.05). Pretreated tattoos responded better to laser treatments than previously untreated tattoos. Subjects felt significantly less pain with PSL than with NSL (P < 0.001). Transient adverse reactions were statistically less pronounced lasting shorter for PSL as for NSL, especially blistering, pruritus, and burning sensation. Hypopigmentation appeared after NSL treatments only, whereas hyperpigmentation was caused by both lasers. No scarring was detected with either laser. CONCLUSIONS: Both laser systems enable acceptable clearance of most tattoos in the present study. PSL cause less collateral skin damage as compared to NSL.

Role of clinical and CT findings in the identification of adult small-bowel intussusception requiring surgical intervention.

BACKGROUND: In adults, intussusception has been considered traditionally to have an underlying aetiology. The aim of this study was to determine CT and clinical features of small-bowel intussusceptions that required surgical intervention. METHODS: Adult patients were identified in whom small-bowel intussusceptions were noted on CT images. The appearance, number, type (enteroenteric versus enterocolic), length and maximum short-axis diameter of intussusceptions, and presence of bowel obstruction (short-axis diameter of proximal small bowel greater than 3 cm) were analysed. The outcome was defined as surgical (complicated) or self-limiting (uncomplicated). Associations between complicated and uncomplicated intussusceptions and patient characteristics were investigated. RESULTS: Among 75 patients (56 male) with a mean age of 45 years, 103 intussusceptions were identified, of which 98 (95 per cent) were enteroenteric and 5 (5 per cent) enterocolic. Only 12 of 103 intussusceptions (12 per cent) in 12 of 75 (16 per cent) patients required surgical therapy and were considered to be complicated, with half of these having a neoplastic lead point. Length (P < 0.001), diameter (P < 0.001) and type (P = 0.002) of intussusception as well as presence of vessels (P = 0.023) within an intussusception on a CT scan, clinical symptoms (P = 0.007) and signs of bowel obstruction (P < 0.001) were associated with a surgical outcome. CONCLUSION: Clinical symptoms, signs of bowel obstruction, type and length of intussusception, and a visible tumour within an intussusception on CT scan were critical signs of complicated intussusception, requiring surgical intervention.

Novel photodynamic coating reduces the bioburden on near-patient surfaces thereby reducing the risk for onward pathogen transmission: a field study in two hospitals.

BACKGROUND: Near-patient surfaces are recognized as a source for hospital-acquired infections. Such surfaces act as reservoirs for microbial contamination by which pathogens can be transmitted from colonized or infected patients to susceptible patients. Routine disinfection of surfaces only results in a temporal elimination of pathogens, and recontamination inevitably occurs shortly between disinfections. AIM: A novel antimicrobial coating based on photodynamics was tested under laboratory conditions and subsequently in a field study in two hospitals under real-life conditions. METHODS: Identical surfaces received a photodynamic or control coating. Bacterial counts [colony-forming units (cfu)/cm2) were assessed regularly for up to 6 months. FINDINGS: The laboratory study revealed a mean reduction of several human pathogens of up to 4.0 ± 0.3 log10. The field study in near-patient environments demonstrated mean bacterial values of 6.1 ± 24.7 cfu/cm2 on all control coatings. Photodynamic coatings showed a significantly lower mean value of 1.9 ± 2.8 cfu/cm2 (P<0.001). When considering benchmarks of 2.5 cfu/cm2 or 5 cfu/cm2, the relative risk for high bacterial counts on surfaces was reduced by 48% (odds ratio 0.38, P<0.001) or 67% (odds ratio 0.27, P<0.001), respectively. CONCLUSION: Photodynamic coatings provide a significant and lasting reduction of bacterial counts on near-patient surfaces, particularly for high bacterial loads, in addition to routine hygiene. The promising results of this proof-of-concept study highlight the need for further studies to determine how this novel technology is correlated with the frequency of hospital-acquired infections.

Incidence and evolution of venous thrombosis during the first 3 months after irreversible electroporation of malignant hepatic tumours.

The incidence and evolution of venous thrombosis adjacent to the ablation zone after percutaneous irreversible electroporation (IRE) were evaluated to identify potential risk factors in patients with hepatic malignancies. 205 venous structures (in 87 patients) within a ≤1.0 cm radius of the ablation zone were assessed after IRE of 112 hepatic lesions (74 primary, 38 secondary hepatic malignancies) by pre-interventional and post-interventional (1-3 days, 6 weeks and 3 months after IRE) contrast-enhanced magnetic resonance imaging. The relationships between venous thrombosis and clinical features were analysed using a binary logistic regression model. In 27 of 87 patients (31%), a total of 67 venous complications were noted during the 3 months follow-up. Thrombosis represented the most frequently observed complication (n = 47; 70.1%), followed by vessel narrowing (n = 20; 29.9%). 5 (10.6%) of 47 thromboses showed spontaneous regression 3 months after IRE. A small vessel diameter (p = 0.011) and post-interventional vessel narrowing (p = 0.006) were independently associated with delayed post-ablative thrombosis. Delayed venous thrombosis frequently occurs after IRE of hepatic malignancies. Pre-existing vessel narrowing and a small vessel diameter represent significant risk factors that require further surveillance and potentially therapeutic intervention.

Laser assisted tattoo removal - state of the art and new developments.

Decorative tattoos including permanent make-up are very popular world-wide. However, some people regret tattooing and seek tattoo removal. Tattooed skin contains numerous solid particles of tattoo pigments. A major mechanism of tattoo removal is laser assisted fragmentation of these particles, which are then transported away from the skin. For many years, Q-switched lasers with nanosecond pulse durations at high light intensities have been applied to cause such fragmentation via rapid heating up while sparing the adjacent tissue. Despite the long-lasting use of such laser treatment, the exact mechanisms of laser assisted fragmentation are hardly investigated. Due to short and intense laser pulses applied, non-linear effects of light (e.g. photoacoustic and optical breakdown) and nonlinear thermal properties in tattoo particles may play a crucial role. In the past few years, lasers with even shorter pulse duration in the sub-nanosecond range were launched for tattoo removal. Theoretical considerations assumed that pulse durations shorter than nanoseconds allow even more effective fragmentation of tattoo particles. A couple of initial studies affirm the effectiveness of picosecond pulses in tattoo treatment, especially in clearing black tattoos. Furthermore, treatment with picosecond lasers seems to be less painful. Consequently, picosecond technology may be a new strategy for more effective removal of tattoo pigments at a lower rate of side effects. But there is an urgent need for more well-designed and randomized controlled trials to compare this treatment modality to the traditional nanosecond technology regarding efficacy and adverse reactions.

Photodynamic inactivation of bacteria to decolonize meticillin-resistant Staphylococcus aureus from human skin.

BACKGROUND: To prevent infections that arise from the skin surface it is necessary to decolonize human skin prior to any proposed treatment or surgical intervention. Photodynamic inactivation of bacteria (PIB) uses cationic photosensitizers that attach to the surface of bacteria, generate reactive oxygen species on light irradiation and thereby kill bacteria via oxidative mechanisms. OBJECTIVES: To evaluate the potential and the safety of PIB for decolonization of bacteria from skin. METHODS: PIB with the new photosensitizer SAPYR [2-((4-pyridinyl)methyl)-1H-phenalen-1-one chloride] was initially tested against different bacterial species in vitro. Then, ex vivo porcine skin samples were used as a model for decolonization of different bacteria species. The numbers of viable bacteria were quantified and the mitochondrial activity of skin cells was histologically analysed (using nitroblue tetrazolium chloride, NBTC). The same procedure was performed for human skin and meticillin-resistant Staphylococcus aureus (MRSA). RESULTS: The in vitro studies showed a 5 log10 reduction of all tested bacterial species. On ex vivo porcine skin samples, PIB reduced the viability of all tested bacterial species by at least 3 log10 steps. On human skin samples ex vivo, PIB reduced the number of viable MRSA by maximal 4·4 log10 steps (1000 µmol L-1 SAPYR, incubation time 10 min, 60 J cm-2 ). NBTC staining showed normal mitochondrial activity in skin cells after all PIB modalities. CONCLUSIONS: The results of this study show that PIB can effectively and safely kill bacteria like MRSA on the skin surface and might have the potential of skin decolonization in vivo.

[Irreversible electroporation (IRE) : A minimally invasive therapeutic option in prostate cancer]. / Irreversible Elektroporation (IRE) : Eine minimal-invasive Methode zur Therapie des Prostatakarzinoms.

CLINICAL PROBLEM: Prostate cancer is one of the most common malignant diseases in older men. As such, screening and early detection are of crucial importance. STANDARD TREATMENT: The standard management of prostate cancer includes radical prostatectomy, radiation therapy, or hormonal therapy. These standard therapies yield excellent oncologic results, but also produce significant side effects. In cases of low-risk prostate cancer, these therapies might result in over-treatment; for this reason, active surveillance has been introduced. However, acceptance of this strategy varies between patients. TREATMENT INNOVATIONS: Irreversible electroporation is a novel non-thermal ablation technique for soft tissues. The ablation mechanism of irreversible electroporation has some theoretical advantages in the treatment of prostate cancer. It allows image-guided focal treatment of malignant prostate tissue and a potential sparing of adjacent structures, thereby theoretically resulting in a marked reduction in treatment-related side effects compared with standard management. PRACTICAL RECOMMENDATIONS: Although irreversible electroporation is a promising ablation technique, it should only be used in the context of clinical trials to treat prostate cancer due to the current lack of solid evidence for this new technology.

Neodymium-doped yttrium aluminium garnet (Nd:YAG) 1064-nm picosecond laser vs. Nd:YAG 1064-nm nanosecond laser in tattoo removal: a randomized controlled single-blind clinical trial.

BACKGROUND: For decades, nanosecond lasers (NSLs) have been used to remove tattoos. Since 2012, pulses of picosecond lasers (PSLs) have been available for tattoo removal. Based on a few observational studies, the claim has been made that PSLs are considerably more effective while showing fewer side-effects in comparison with NSLs. OBJECTIVES: To compare the efficacy and side-effects of a PSL side by side with an NSL for tattoo removal. METHODS: Twenty-one patients with 30 black tattoos were treated with PSL and NSL in a split-study design in two sessions at intervals of 6 weeks. The safety and efficacy of laser treatments were determined by blinded observers assessing randomized digital photographs in this prospective clinical study. The primary end point was the clearance of the tattoos ranging in quartiles from 0% to 100%; secondary end points were side-effects and pain. RESULTS: The average clearance overall as evaluated showed no statistical difference between NSL and PSL (P = 1·00). Using a visual analogue scale (0 = no pain, 10 = maximum pain), a value of 3·8 ± 1·0 was reported for the PSL, which was statistically different from NSL (7·9 ± 1·1, P < 0·001). Transient side-effects were observed, as well as hypo- and hyperpigmentation, but there was no statistically significant difference between PSL and NSL. CONCLUSIONS: After two treatments of black tattoos with a neodymium-doped yttrium aluminium garnet laser (1064 nm), the use of picosecond pulses does not provide better clearance than nanosecond pulses. However, pain is less severe when using a PSL.

Indocyanine green-augmented diode laser therapy vs. long-pulsed Nd:YAG (1064 nm) laser treatment of telangiectatic leg veins: a randomized controlled trial.

BACKGROUND: Telangiectatic leg veins (TLV) represent a common cosmetic problem. Near infrared lasers have been widely used in treatment because of their deeper penetration into the dermis, but with varying degrees of success, particularly because of different vessel diameters. Indocyanine green (ICG)-augmented diode laser treatment (ICG+DL) may present an alternative treatment option. OBJECTIVES: This trial evaluates the efficacy of ICG+DL in the treatment of TLV and compares the safety and efficacy of therapy with the standard treatment, the long-pulsed neodymium-doped yttrium aluminium garnet (Nd:YAG) laser. METHODS: In a prospective randomized controlled clinical trial, 29 study participants with TLV were treated with a Nd:YAG laser (λem = 1064 nm, 160-240 J cm(-2) , 65-ms pulse duration, 5-mm spot size) and ICG+DL (λem = 810 nm, 60-110 J cm(-2) , 48-87-ms pulse duration, 6-mm spot size; total ICG dose 4 mg kg(-1) ) in a side-by-side comparison in one single treatment setting that included histological examination in four participants. Two blinded investigators and the participants assessed clearance rate, cosmetic appearance and adverse events up to 3 months after treatment. RESULTS: According to both the investigators' and participants' assessment, clearance rates were significantly better after ICG+DL therapy than after Nd:YAG laser treatment (P < 0·05). On a 10-point scale indicating pain during treatment, participants rated ICG+DL therapy to be more painful (6·1 ± 2·0) than Nd:YAG laser (5·4 ± 2·0). CONCLUSIONS: ICG+DL therapy represents a new and promising treatment modality for TLV, with high clearance rates and a very good cosmetic outcome after one single treatment session.

Weather conditions and daylight-mediated photodynamic therapy: protoporphyrin IX-weighted daylight doses measured in six geographical locations.

BACKGROUND: Photodynamic therapy (PDT) is an attractive therapy for nonmelanoma skin cancers and actinic keratoses (AKs). Daylight-mediated methyl aminolaevulinate PDT (daylight-PDT) is a simple and painless treatment procedure for PDT. All daylight-PDT studies have been performed in the Nordic countries. To be able to apply these results in other parts of the world we have to compare the daily protoporphyrin IX (PpIX) light dose in other countries with the PpIX light doses found in Nordic countries. OBJECTIVES: To calculate where and when daylight-PDT of AKs was possible in six different geographical locations using ground stations measuring PpIX-weighted daylight doses. METHODS: PpIX-weighted daylight doses were measured using a dosimeter with a customer-specific photodiode with a detector sensitivity that mimics the PpIX absorption spectrum and measures in 'PpIX doses'. The dosimeters were built into ground stations that were placed in six geographical locations measuring from July to December 2008. Temperature data for each location were obtained from the internet. The maximal ultraviolet (UV) index for Copenhagen was obtained for the measuring period of the dosimeters. RESULTS: If the PpIX light dose should be above 8Jcm(-2) and the maximum temperature of the day at least 10°C, it was possible to treat patients on nearly all days until the middle of September in Reykjavik and Oslo, until the last week of October in Copenhagen and Regensburg, until the middle of November in Turin and all year in Israel. CONCLUSIONS: Where and when to perform daylight-PDT depends on the PpIX light dose and outdoor temperature. The PpIX light dose was influenced by the geographical location (latitude), weather condition and time of year. The UV index was not more suitable than temperature and weather to predict if the intensity of daylight would be sufficient for daylight-PDT.

Intravenous injection of indocyanine green to enhance laser-assisted coagulation of blood vessels in skin - an animal study.

BACKGROUND: Laser therapy of vascular lesions, such as port wine stains (PWS) or leg veins are still imperfect due to different diameters and depth of vessels in tissue. We propose to improve blood vessel coagulation by intravenous introduction of an exogenous chromophore (indocyanine green, ICG) that effectively converts near-infrared (NIR) laser light into heat. OBJECTIVE: The purpose of this study was to determine the plasma clearance rate, systemic toxicity and histological effects of ICG-assisted laser therapy in an animal model. METHODS: Piglets received intravenous injection of ICG. Blood samples were collected at different times. Systemic toxicity was assessed by measuring liver enzyme levels and other indicators of liver function. The plasma clearance rate of ICG was determined by light absorption measurement in blood samples. The skin was irradiated with a diode laser (810 nm) using radiant exposures from 31 to 80 J/cm². Skin reaction at the treatment site was graded, and punch biopsies were taken for histological examination at 24 and 72 h after treatment. RESULTS: No hepatic toxicity was observed. The clinical examination revealed no adverse skin reactions at 24 or 72 h after laser irradiation. This was confirmed by histological evaluation that showed efficient vessel coagulation without damage of the epidermis or dermis. CONCLUSIONS: In light of these in vivo results, we suggest that ICG-assisted laser therapy could substantially improve clinical outcomes of PWS or leg veins treatment with minimal risk of adverse reactions.

Indocyanine green-augmented diode laser treatment of port-wine stains: clinical and histological evidence for a new treatment option from a randomized controlled trial.

BACKGROUND: Complete clearance of port-wine stains (PWS) is difficult to achieve, mainly because of the resistance of small blood vessels to laser irradiation. Indocyanine green (ICG)-augmented diode laser treatment (ICG+DL) may overcome this problem. OBJECTIVES: To evaluate the feasibility of ICG+DL therapy of PWS and to compare the safety and efficacy of ICG+DL with the standard treatment, flashlamp-pumped pulsed dye laser (FPDL). METHODS: In a prospective randomized controlled clinical study, 31 patients with PWS were treated with FPDL (λ(em)=585 nm, 6 J cm(-2) , 0.45 ms pulse duration) and ICG+DL (λ(em)=810 nm, 20-50 J cm(-2) , 10-25 ms pulse duration, ICG-concentration: 2 mg kg(-1) body weight) in a split-face modus in one single treatment setting that included histological examination (haematoxylin and eosin, CD34). Two blinded investigators and the patients assessed clearance rate, cosmetic appearance and side-effects up to 3 months after treatment. RESULTS: ICG+DL therapy induced photocoagulation of medium and large blood vessels (>20 µm diameter) but not of small blood vessels. According to the investigators' assessment, clearance rates and cosmetic appearance were better after ICG+DL therapy than after FPDL treatment (P=0.114, P=0.291, respectively), although not up to a statistically significant level, whereas patients considered these parameters superior (P=0.003, P=0.006, respectively). On a 10-point scale indicating pain during treatment, patients rated ICG+DL to be more painful (5.81 ± 2.12) than FPDL treatment (1.61 ± 1.84). CONCLUSION: ICG+DL represents a new and promising treatment modality for PWS, but laser parameters and ICG concentration need to be further optimized.

The decrease of pigment concentration in red tattooed skin years after tattooing.

BACKGROUND: Tattooing entails the injection of high amounts of colourants into skin. Excepting black inks, red azo pigments are the most frequent colourant used. Part of the pigment is transported away via lymphatic system. Another part can be decomposed in skin, which might be responsible for many known adverse skin reactions. OBJECTIVE: The aim of this study was to estimate the extent of decomposition and transportation by measuring the decrease of pigment concentration in human skin under in vivo conditions. METHODS: Red pigments were extracted from nine tattooed skin specimen and attempted quantification by using HPLC technology. To optimize quantification, we synthesized five common red azo pigments with purity at 98% and used them as HPLC reference substances. RESULTS: In five of the nine skin specimens, we were able to identify and subsequently to quantify the red tattoo pigments such as Pigment Red 22 or Pigment Red 112. The mean pigment concentration in skin was 0.077 ± 0.046 mg/cm². As the pigment concentration in skin ranges from 0.60 to 9.42 mg/cm² (mean: 2.53) directly after tattooing, we estimate a decrease of 87 to 99% of pigment concentration in skin after tattooing. CONCLUSION: Millions of people have many and large tattoos, whereas a single tattoo frequently covers a skin area of more than 300 cm². Thus, the major part of more than 760 mg of azo pigments either decomposes in skin or migrates in the body. That may pose a health risk on tattooed individuals, in particular may cause severe skin reactions.

Recurring mistakes in tattoo removal. A case series.

In the Western world, more than 10% of the population have at least 1 tattoo. If the tattoo is removed, the tattoo pigment particles in the skin can be selectively destroyed by means of selective photothermolysis. This treatment requires laser pulses of short durations (nanoseconds) and high intensities. We report on 12 patients who received treatments with improper treatment parameters. In all patients, we diagnosed hypo- or hyperpigmentations and scar formation at the treatment site. In particular, the pulse duration of the light sources or lasers applied were considerably longer than those required by the principles of selective photothermolysis. The light intensities of those devices are normally not sufficient to destroy the pigment particles. Instead of destruction, the pigment particles in the skin are heated up and the heat is conducted to the adjacent tissue causing unspecific tissue injury. Lasers or intense pulsed light sources with millisecond pulses and low light intensities are clearly not suitable to be applied for tattoo removal.

In vivo relevance for platelet glycoprotein Ibalpha residue Tyr276 in thrombus formation.

BACKGROUND: Platelet glycoprotein (GP) Ib-IX-V supports platelet adhesion on damaged vascular walls by binding to von Willebrand factor (VWF). For several decades it has been recognized that the alpha-subunit of GP (GPIbalpha) also binds thrombin but the physiological relevance, if any, of this interaction was unknown. Previous studies have shown that a sulfated tyrosine 276 (Tyr276) is essential for thrombin binding to GPIbalpha. OBJECTIVES: This study investigated the in vivo relevance of GPIbalpha residue Tyr276 in hemostasis and thrombosis. METHODS: Transgenic mouse colonies expressing the normal human GPIbalpha subunit or a mutant human GPIbalpha containing a Phe substitution for Tyr276 (hTg(Y276F)) were generated. Both colonies were bred to mice devoid of murine GPIbalpha. RESULTS: Surface-expressed GPIbalpha levels and platelet counts were similar in both colonies. hTg(Y276F) platelets were significantly impaired in binding alpha-thrombin but displayed normal binding to type I fibrillar collagen and human VWF in the presence of ristocetin. In vivo thrombus formation as a result of chemical damage (FeCl(3)) demonstrated that hTg(Y276F) mice have a delayed time to occlusion followed by unstable blood flow indicative of embolization. In models of laser-induced injury, thrombi developing in hTg(Y276F) animals were also less stable. CONCLUSIONS: The results demonstrate that GPIbalpha residue Tyr276 is physiologically important, supporting stable thrombus formation in vivo.

Hereditary haemorrhagic telangiectasia treated by pulsed neodymium:yttrium-aluminium-garnet (Nd:YAG) laser (1,064 nm).

Hereditary haemorrhagic telangiectasia (HHT) is a familial, autosomal, dominant, multi-system, vascular, dysplasia. Besides repetitive epistaxis, cutaneous eruptive macules and nodules lead to recurring bleeding and cosmetic problems. We report on a pilot study of four cases of HHT in which cutaneous lesions were treated with a pulsed neodymium:yttrium-aluminum-garnet (Nd:YAG) laser (1,064 nm). Pulsed Nd:YAG laser treatment, without anaesthesia, was performed several times on eruptive angiomas on palmar and facial skin. Lesions on fingers and face mostly showed very good, or even complete, clearing after the first laser treatment. Several macules required multiple treatment; only a few lesions showed no effect. Pulsed Nd:YAG laser therapy (1,064 nm) appears to be an effective and safe treatment option for hereditary haemorrhagic telangiectasia on the skin of face and extremities.