Pigment Spread Research Articles

Melanosis coli as a factor in the development of colorectal cancer

Melanosis coli is characteristic of patients with a long history of constipation and the use of laxatives – anthraquinone derivatives, which are quite prescribed by doctors and patients independently. As a rule, melanosis develops after 4–9 months of taking drugs from this group and is most common in women in the older age group. Pathogenetically, melanosis is characterized by the deposition of pigment in the lamina propria of the mucous membrane, manifested by a pronounced dark coloration of the mucosa, depending on the degree of pigmentation of the colon, melanosis is divided into three degrees. The most common localization is in the right colon, but cases with the spread of pigmentation to the left colon and other parts have been described. Diagnosis does not cause difficulties, melanosis coli is detected during routine endoscopic examination, and is confirmed by pathomorphological examination. Melanosis is an actual problem in terms of the risk of developing colorectal cancer (CRC). There are already convincing data proving that the frequency of detection of adenomas increases not only due to the absence of pigment accumulation by them, but also the pathogenetic aspects of the influence of melanosis on the occurrence and development of adenomas have been described. Proteomic analysis allows to determine the expression of various proteins, which confirms the role of melanosis in the development of colorectal cancer. It is also interesting to note that sometimes there are some difficulties in the differential diagnosis of melanosis, for example, with ischemic lesions of the colon. Patients with melanosis coli require close attention, timely diagnosis and treatment.

Comparison of micro skin grafting and transplantation of non-cultured melanocyte keratinocyte suspension for the treatment of stable vitiligo: A pilot study

Objectives: We conducted this pilot study to compare the outcomes of non-cultured epidermal suspension (NCES) with that of micro skin grafting (MSG) in the treatment of stable vitiligo Material and Methods: Twenty-nine patients with clinically stable vitiligo lesions (defined as the occurrence of no new lesions and no increase in the size of preexisting lesions for the past six months) and age group between 12 and 70 years were enrolled in the study. The enrolled patients were randomized into two groups using simple randomization using computer-generated random numbers, and allocation concealment was done using opaque sealed envelopes. Group 1 was comprised of 15 patients with 23 stable vitiligo lesions, and group 2 comprised of 14 patients with 22 stable vitiligo lesions. Patients in group 1 were transplanted with non-cultured melanocytes and keratinocytes. Patients in group 2 were treated using micro skin grafts after obtaining written informed consent. Ethical clearance was obtained from the Institute ethics committee, and the principles of the 1975 Declaration of Helsinki were followed. Results: At the end of four months post-treatment, two patients (one in each group) did not follow up after removal of dressing at the first week. Results were analyzed at four months in 27 patients – 14 patients with 22 lesions in group 1 and 13 patients with 21 lesions in group 2. Repigmentation at 16 weeks post-surgery was evaluated. Excellent repigmentation (>90%) was seen in 45.45% of lesions in the non-cultured epidermal suspension (NCES) group as compared to 38.09% of lesions in the micro skin grafting (MSG) group, and this difference was not statistically significant (p = 0.7597). Repigmentation > 75% was achieved in 54.54% of lesions in the non-cultured epidermal suspension (NCES) group compared to 38.09% of lesions in the micro skin grafting (MSG) group, and this difference was not statistically significant (p = 0.3640). Both the groups did not have any significant complications like scarring, milia, or any cobble stoning at the donor site. The recipient area had resistance to the spread of pigment at the margins of lesions. Six lesions in both the groups with excellent response, 3 in each group had achromic fissure or hypopigmented halos at the margins of lesions. Conclusion: Both non-cultured epidermal suspension (NCES) and micro skin grafting (MSG) have been found to be safe and effective modalities with comparable efficacy in the surgical treatment of stable vitiligo. However, future trials on large sample sizes are warranted to validate our results.

Metallic glass coating for improved needle tattooing performance in reducing trauma: analysis on porcine and pig skins

The dissemination of tattooing into mainstream culture has raised concerns pertaining to the medical implications of these practices. This paper reports on the coating of tattoo needles with metallic-glass (MG) to reduce trauma to the skin. Extensive experimentation using pork samples and live pigs demonstrated the beneficial effects of non-stick MG coatings. Following 30 insertions into pork skin, significantly less tissue adhered to the MG-coated needles than to uncoated needles. MG-coated needles were also shown to reduce the spread of pigment to the surface of surrounding skin by up to 57%. This resulted in narrower tattoo lines of higher density, indicating that MG-coated needles could be useful in high-resolution tattooing. Histopathological analysis on live pigs revealed severe trauma induced by bare needles, as indicated by the secretion of fluids immediately after tattooing. The wounds formed by coated needles closed within 2 h after tattooing; however, those formed by bare needles remained open for at least 2 h and inflammation was still observed after 3 days. At 5 days after tattooing, skin punctured by the coated needle was entirely healed, whereas skin punctured by the bare needle was still covered with scabs. In addition to the medical benefits, it appears that MG-coated needles could improve the quality of tattoos, based on the fact that the amount of pigment retained in the skin is inversely proportional to the trauma caused by needles.

Treatment of idiopathic guttate hypomelanosis with 5-fluorouracil tattooing using a handheld needle

Idiopathic guttate hypomelanosis is a common acquired skin condition characterized by multiple discrete, round, porcelain-white macules on the extensor aspects of the extremities in the elderly population. Most patients are reassured of the benign nature of the condition and no treatment is recommended. Despite the benignity, many patients seek treatment because the condition is cosmetically displeasing. There are several topical, physical, and surgical modalities available for idiopathic guttate hypomelanosis, but the results are variable and unpredictable, and there is no standardization of the treatment regimens. Treatment modalities such as cryotherapy and spot peels are frequently associated with dyspigmentation, especially in darker skin types. We sought to develop a simple office-based procedure to treat idiopathic guttate hypomelanosis with minimal adverse effects, providing consistent results, and which can be performed by using readily available instruments (eg, needle) even in a remote or resource-poor setting. Topical anesthesia (eutectic mixture of lignocaine 2.5% weight by weight and prilocaine 2.5% weight by weight cream) was applied on lesions for 45 minutes. A simple 24-gauge hypodermic needle was dipped in a vial containing a solution of 5-fluorouracil at 50 mg/mL. The needle was filled by capillary action and the tip of the needle was later inserted in the lesion to create mild pinpoint bleeding under aseptic conditions. 5-Fluorouracil invokes a regulatory response within the fibroblasts of the papillary dermis, which causes regulation of melanocytes.1Wambier C.G. Wambier SPdF. Pereira Soares M.T. Breunig J. Cappel M.A. Landau M. 5-Fluorouracil tattooing for idiopathic guttate hypomelanosis.J Am Acad Dermatol. 2018; 78: e81-e82Abstract Full Text Full Text PDF PubMed Scopus (8) Google Scholar,2Arbache S. Roth D. Steiner D. et al.Activation of melanocytes in idiopathic guttate hypomelanosis after 5-flourouracil infusion using a randomized, split-body, single blinded, placebo controlled clinical trial.J Am Acad Dermatol. 2018; 78: 212-215Abstract Full Text Full Text PDF PubMed Scopus (16) Google Scholar 5-Fluorouracil makes a long-lasting favorable microenvironment for the melanocyte migration and subsequent pigment spread. We conducted a total of 2 sittings 1 month apart, which yielded complete uniform repigmentation (Fig 1, A and B). No treatment-related adverse effects were observed during or after the procedure. We tried a simple cost-effective treatment that can be easily performed and obviates the need for any high-end or complex equipment or instruments to treat idiopathic guttate hypomelanosis. This is an off-label use of 5-fluorouracil and the safety and efficacy remain unknown. The novelty of our pearl is that the previously reported description used an actual tattooing device, but we are simply using a needle available in any office. Treatment of idiopathic guttate hypomelanosis with a tattoo device versus a handheld needleJAAD InternationalVol. 3PreviewTo the Editor: We were pleased to read the case report by Rambhia and Rambhia describing the use of a hypodermic needle dipped in 5-fluorouracil to treat a patient with idiopathic guttate hypomelanosis (IGH).1 It is a simple and original solution to treat challenging dermatosis. Full-Text PDF Open Access

Prediction of response to surgical therapy in nonsegmental vitiligo based on an immunohistochemical cutoff value

Background Objective assessment of stability is still lacking in nonsegmental vitiligo (NSV).Objective To suggest immunohistochemical cutoff values that can predict a good response to surgical therapy of NSV cases.Patients and methods Fifteen active and 15 stable NSV cases were included in the present study. A 3-mm perilesional biopsy from perilesional skin was obtained for immunohistochemical staining for CD8, CD45RO, and FoxP3. Mini punch graft was done for all cases, and response was assessed after 6 months. Good responders had spread of pigment more than 1 mm, and poor responders had no spread of pigment or loss of color. Comparison of immunohistochemical findings according to response to surgery was done, and a cutoff value was calculated when feasible.Results Ten active and 10 stable cases continued 6 months of follow-up. Loss of color occurred in all active cases, whereas five patients had no spread and five patients had good response in the stable group. Good responders had a significantly lower level of CD8+ (P=0.001) and CD45RO+ cells (P=0.020), with cutoff values of 7 and 11 cells/high-power field, respectively. No significant difference was found in FoxP3+ cells.Conclusion A suggested cutoff value of 7 CD8+ T cells and 11 CD45RO+ cells/high-power field can predict a good response to surgery in NSV.

Melanonychia striata: clarifying behind the Black Curtain. A review on clinical evaluation and management of the 21st century.

Melanonychia striata is characterized by a tan, brown, or black longitudinal streak within the nail plate that runs from the proximal nail fold to the distal part of the nail plate. Melanonychia striata is due to increased activity of melanocytes or melanocytic hyperplasia in the nail matrix with subsequently increased melanin deposition in the nail plate. The most common cause of melanonychia striata associated with melanocytic activation is ethnic melanonychia which occurs in dark-skinned individuals. Other causes of melanonychia striata related to melanocytic activation include pregnancy, chronic local trauma, infections, medications, dermatological disorders, endocrine disorders, alkaptonuria, hemochromatosis, porphyria, graft-vs-host disease, Peutz-Jeghers syndrome, and Laugier-Hunziker syndrome. Causes of melanonychia striata associated with melanocytic hyperplasia include nail matrix melanocytic nevus, nail lentigo, and nail apparatus/subungual insitu and invasive melanoma. In most cases, melanonychia striata is a benign condition, especially in children. Consequently, most investigators advocate a wait-and-see approach. Nail apparatus/subungual melanoma should be suspected if there is an abrupt onset after middle age, personal or family history of melanoma, rapid growth, darkening of a melanonychia band, pigment variegation, blurry lateral borders, irregular elevation of the surface, a bandwidth >3mm, proximal widening, associated nail plate dystrophy, single rather than multiple digit involvement, and periungual spread of pigmentation onto the adjacent cuticle and/or proximal and/or lateral nail folds (Hutchinson sign). Prolonged follow-up is mandatory for early detection of possible malignant changes.

Management of gingival hyperpigmentation by the surgical abrasion: A case report

Health and appearance of gingiva are important parts of a smile. The spread of pigmentation depends on melanoblastic activity. The gingival color plays a role in esthetics. Diff erent treatment modalities have been reported for depigmentation of gingiva such as bur abrasion, partial thickness fl ap, cryotherapy, electrosurgery and laser. In this case, depigmentation was done with bur abrasion, which is simple, eff ective and yields good results, along with good patient satisfaction.

Autologous mini punch grafting: an experience of using motorized power punch in 10 patients.

Background:Autologous mini punch grafting (MPG) is a safe, effective and easy technique that can be performed on any site with minimal side effects and good cosmetic results. Large areas of stable generalised vitiligo require more grafts and are time consuming. Hence multiple sessions of surgery need to be scheduled. We share our experience of using motorised power punches to increase the speed of surgery in large areas of stable vitiligo in 10 patients.Materials and Methods:Ten patients in the age group of 12-55 years were treated with miniature punch grafting using power punches in single session on various sites. The power punches of 1-1.5 mm diameter were used to score donor and recipient sites, either of same or less than 0.2-0.3 mm size punches. The harvested grafts from donor site were then secured in the recipient beds and dressed.Results:The average number of grafts harvested per session was 125-185, the duration of surgery ranged from 45 to 90 minutes. Perigraft pigment spread was seen at 3 weeks. Complete repigmentation was observed in 3-4 months in eight patients. Cobble stoning was observed in one patient, and all donor sites healed well with superficial scarring.Conclusion:We conclude that autologous MPG with motorised power punches for stable vitiligo, especially on large areas including difficult sites can be performed with ease in comparatively lesser time in a single session, greatly benefiting the patients.

Target-like Pigmentation After Minipunch Grafting in Stable Vitiligo.

Surgical treatment for vitiligo has been ever evolving. Each surgical modality has its own benefits and limitations. Miniature punch grafting is the most extensively performed surgery, which gives good results in stable vitiligo. Herein we report an unusual type of repigmentation observed after minipunch grafting in a patient of stable vitiligo, which resembled target-like lesions with a “perigraft halo” surrounding individual grafts. Such pigment spread occurred despite the use of 0.5 mm larger graft from the donor site.

Comparative Study of Flip-top Transplantation and Punch Grafting in Stable Vitiligo

Because of limitations of medical treatment in vitiligo, various surgical techniques have been devised. There is a scarcity of comparative studies of surgical techniques in vitiligo. To compare the effectiveness and side-effect profile of flip-top transplantation (FTT) and punch grafting (PG) in patients with stable vitiligo. Twenty patients with stable vitiligo were treated using PG and FTT on 26 lesions; 156 grafts were taken (78 PG, 78 flip-top grafts). They were graded for response in the form of repigmentation as excellent (91-100%), very good (76-90%), good (51-75%), fair (31-50%), poor (<30%). Onset and maximum pigment spread were observed. Sixty-five percent of our patients showed excellent repigmentation with FTT, versus 50% with PG (p=.44). Thirty percent of patients with FTT and 40% with PG had side effects. The differences in onset of repigmentation, completion, and pigment spread between the two techniques were statistically significant (p<.001). Flip-top transplantation was equally effective as PG for treating vitiligo, although graft uptake rate was higher with FTT, with more pigment spread, and cost less.

Possible mechanisms by which topical 5-Fluorouracil and dermabrasion could induce pigment spread in vitiligo skin: an experimental study.

The combination of skin ablation and 5-Fluorouracil (5-FU) ointment was previously tried in the treatment of vitiligo, and good results were specifically reported in glabrous skin without follicular melanocyte reservoirs. Methods. This study was carried out on the skins of seven guinea pigs: three were treated with mechanical dermabrasion plus topical 5-FU in an achromic area contiguous to a pigmented area; two were treated by only dermabrasion in a similar area; and two were treated by topical 5-FU alone. Clinical, histological, and ultrastructural studies were performed over two months. Results. In guinea pigs treated with dermabrasion plus 5-FU, we observed firstly a delay of wound healing with an obvious inflammatory reaction, and, after two months, evident pigment spread from the pigmented into the achromic area. After six months, we noticed black hair regrowing in the achromic area. Pigment spread was not seen in the guinea pigs skin treated by either dermabrasion or topical 5-FU. We suggest that the inflammatory mediators and enzymes (metalloproteinases), which are locally released over a long time, could stimulate and facilitate melanocyte proliferation and migration through the enlarged intercellular spaces of the epidermis. This sequence of events may be applied to vitiligo patients treated with 5-FU on ablated lesions.

Efficacy of narrowband ultraviolet B versus excimer radiation in repigmenting vitiligo after minigrafting on the distal arms

To the Editor: Vitiligo, a common pigmentary disorder, is often difficult to treat. Phototherapy, including narrowband ultraviolet (UV) B (NBUVB) radiation, psoralen and UVA radiation, and the 308-nm excimer laser, is often successful; however, acral lesions are usually resistant to treatment.1Njoo M.D. Spuls P.I. Bos J.D. Westerhof W. Bossuyt P.M. Nonsurgical repigmentation therapies in vitiligo: meta-analysis of the literature.Arch Dermatol. 1998; 134: 1532-1540Crossref PubMed Scopus (236) Google Scholar, 2Passeron T. Ortonne J.-P. Use of the 308-nm excimer laser for psoriasis and vitiligo.Clin Dermatol. 2006; 24: 33-42Abstract Full Text Full Text PDF PubMed Scopus (57) Google Scholar A new 308-nm monochromatic excimer light (MEL) device (Excilite, National Biological Corp, Beachwood, OH) has been developed for the treatment of vitiligo that may be more effective in achieving repigmentation compared with older modalities.3Casacci M. Thomas P. Pacifico A. Bonnevalle A. Paro Vidolin A. Leone G. Comparison between 308-nm monochromatic excimer light and narrowband UVB phototherapy (311–313 nm) in the treatment of vitiligo–a multicenter controlled study.J Eur Acad Dermatol Venereol. 2007; 21: 956-963Crossref PubMed Scopus (93) Google Scholar We performed a pilot study to determine the efficacy of this device for acral vitiligo and its effect on punch grafts compared with standard phototherapy. Thirteen patients gave written informed consent for this study, which was approved by a local institutional review board. Patients had stable vitiligo on the forearms and hands, defined as no more than 10% reported change in lesion size over the previous 6 months, lack of improvement with topical immunomodulators or corticosteroids for 6 months, and not receiving more than 12 previous phototherapy treatments. Five 1.5-mm minigrafts were harvested from hip skin and placed in each of two target lesions per patient. One week after transplantation, the MEL device was used on one target lesion, while the other was treated with a hand-foot NBUVB device (Hand/Foot II NBUVB system, National Biological Corp) 3 times weekly for 12 weeks. Treatment assignments were based on a computer-generated randomization code. Initial dose was 200 mJ on both sides with an increase of 15% per treatment on the NBUVB side and 50 mJ on the MEL device side. The primary outcome measure was improvement in target lesion size performed by a blinded investigator using Scion Image J measurement software (National Institutes of Health, Bethesda, MD) on baseline and week-12 images. These images were taken under standardized conditions. The majority of the patients were Hispanic (61%) and had skin type III (46%) or IV (31%), mean age of 47 years (range 21-74 years), and mean duration of vitiligo of 15 years (range 4-44 years). Ten patients completed the trial. The remaining 3 did not have any adverse effects before dropping out. NBUVB gave a significant improvement (median = 13%, Wilcoxon P = .037) at week 12 (Fig 1) whereas the MEL device did not show significant improvement (median improvement 9%, Wilcoxon P = .169). There was no significant difference between standard NBUVB and the MEL device over time (baseline to 12 weeks) using multivariate repeated measures analysis of variance. Of the 37% of grafts surviving at week 12, maximum pigment spread was a mean 4.5 times the original graft size with NBUVB versus 2 times original graft size with the MEL device. This difference was not significant (Fig 2). There were no adverse events.Fig 2Vitiligo in patient 7 at 1 week after grafting (A) and showing improvement at week 12 after standard narrowband ultraviolet B therapy (B).View Large Image Figure ViewerDownload Hi-res image Download (PPT) This study confirms the difficulty of treating vitiligo in acral locations. Even after 12 weeks and 36 treatment sessions, target lesions improved only 9% to 13% with the MEL device, which was not better than standard NBUVB therapy. Maximum pigment spread from grafts did not show a significant difference either. Although this new device shows promise in other locations, acral vitiligo was relatively unresponsive.

First Report of Cobweb on White Button Mushroom (Agaricus bisporus) in Spain Caused by Cladobotryum mycophilum.

Between 2008 and 2011, symptoms of cobweb were observed in commercial white button mushroom (Agaricus bisporus) crops in Castilla-La Mancha (Spain). Typical symptoms started as white, cobweb-like mycelial growth over the surface of the casing soils and fruiting bodies. Later, the mycelium changed to a grayish white, dense powder and the affected fruiting bodies turned pale yellow or reddish brown before rotting. Two types of cap spotting were observed, dark brown spots with a poorly defined edge and light brown spots. The first symptoms were commonly seen in the second or third break (flush) of mushrooms. Infected tissues of A. bisporus were plated onto potato dextrose agar (PDA) and a parasitic fungus was isolated. Fungal colonies consisted of abundant, cottony, aerial mycelium spreading rapidly over the PDA, and red pigment spreading into the agar. The cultures lacked a camphor odor. Conidiogenous cells were 24 to 45 μm long, 3 to 6 μm wide basally, and tapered slightly to the tip. Conidia were cylindrical to narrowly ellipsoidal, 15 to 28 × 8 to 11 μm, and zero- to three-septate. Total DNA was extracted and the internal transcribed spacer (ITS) region of rDNA amplified for one mycelial isolate using ITS1F/ITS4 primers (2,4). The amplicon was sequenced (GenBank Accession No. JQ004732). BLAST analysis showed highest similarity (99 and 100%) of the ITS sequence to four ITS sequences of Cladobotryum mycophilum (teleomorph Hypomyces odoratus) (GenBank Accession Nos. AB527074, JF505112, Y17095, and Y17096) (1,3) among other sequences of the same species. Two pathogenicity trials (A and B) were performed in mushroom-growing rooms, with 24 blocks in each assay containing pasteurized, spawned, and incubated A. bisporus substrate (10 kg, 0.15 m2). The blocks were cased with a 35-mm layer of a peat-based casing soil (5.5 liter/block). Nine days after casing, a conidial suspension (7.5 × 103 conidia/ml) of one isolate of C. mycophilum was sprayed (20 ml/block) onto the surface of the casing layer of 12 blocks at 106 conidia/m2. Twelve blocks were sprayed with sterile distilled water as a control treatment. Blocks were maintained at 17.5°C and 90% relative humidity. The first cobweb symptoms developed 25 days after inoculation, between the second and third breaks in trial A; and after 11 days, between the first and second breaks in trial B. C. mycophilum was consistently reisolated from eight inoculated blocks (67%) in trial A, and 11 inoculated blocks (92%) in trial B. The total area of the crop affected by cobweb was 30% in inoculated blocks in trial A and 45% in trial B. The noninoculated blocks remained healthy. Compared with the noninoculated control blocks, a 10.7% decrease in yield of mushrooms was observed in trial A and 9.1% in trial B. Previously, C. dendroides was the only known causal agent of cobweb in Spain. To our knowledge, this is the first report of C. mycophilum causing cobweb in white button mushroom in Spain, although the disease and causal agent were previously reported on cultivated king oyster mushroom (Pleurotus eryngii) in Spain (3).

Environmental stresses of field growth allow cinnamyl alcohol dehydrogenase-deficient Nicotiana attenuata plants to compensate for their structural deficiencies.

The organized lignocellulosic assemblies of cell walls provide the structural integrity required for the large statures of terrestrial plants. Silencing two CINNAMYL ALCOHOL DEHYDROGENASE (CAD) genes in Nicotiana attenuata produced plants (ir-CAD) with thin, red-pigmented stems, low CAD and sinapyl alcohol dehydrogenase activity, low lignin contents, and rubbery, structurally unstable stems when grown in the glasshouse (GH). However, when planted into their native desert habitat, ir-CAD plants produced robust stems that survived wind storms as well as the wild-type plants. Despite efficient silencing of NaCAD transcripts and enzymatic activity, field-grown ir-CAD plants had delayed and restricted spread of red stem pigmentation, a color change reflecting blocked lignification by CAD silencing, and attained wild-type-comparable total lignin contents. The rubbery GH phenotype was largely restored when field-grown ir-CAD plants were protected from wind, herbivore attack, and ultraviolet B exposure and grown in restricted rooting volumes; conversely, it was lost when ir-CAD plants were experimentally exposed to wind, ultraviolet B, and grown in large pots in growth chambers. Transcript and liquid chromatography-electrospray ionization-time-of-flight analysis revealed that these environmental stresses enhanced the accumulation of various phenylpropanoids in stems of field-grown plants; gas chromatography-mass spectrometry and nuclear magnetic resonance analysis revealed that the lignin of field-grown ir-CAD plants had GH-grown comparable levels of sinapaldehyde and syringaldehyde cross-linked into their lignins. Additionally, field-grown ir-CAD plants had short, thick stems with normal xylem element traits, which collectively enabled field-grown ir-CAD plants to compensate for the structural deficiencies associated with CAD silencing. Environmental stresses play an essential role in regulating lignin biosynthesis in lignin-deficient plants.

First Report of Cladobotryum mycophilum Causing Cobweb on Cultivated King Oyster Mushroom in Spain.

In 2010, symptoms of cobweb were observed on cultivated king oyster mushroom (Pleurotus eryngii) in Castilla-La Mancha (Spain) affecting 16% of the blocks of substrate cultivated. Cobweb appeared at the end of the crop cycle, first as small, white patches on the casing soil, subsequently spreading to the nearest king oyster mushroom by means of a fine gray-white mycelium, and eventually sporulating to produce masses of dry spores. The mycelium can quickly cover pinheads, stalks, pileus, and gills, eventually resulting in decomposition of the entire fruit body. Infected tissues of P. eryngii were plated onto potato dextrose agar (PDA) and the parasitic fungus was isolated. Fungal colonies consisted of abundant and cottony aerial mycelium spreading rapidly on PDA and red pigment spreading in the agar. Conidiogenous cells were 24 to 35 μm long, 3.5 to 5 μm wide basally, and tapered slightly to the tip. Conidia were cylindrical to narrowly ellipsoidal, 17 to 25 (-28) × 8 to 10 μm, and zero to three septate. Total DNA was extracted and the internal transcribed spacer (ITS) region of rDNA was amplified for one isolate using ITS1F/ITS4 primers (1,3). The amplicon was sequenced (GenBank Accession No. JF505112). BLAST analysis showed 100% similarity of the obtained ITS sequence with two sequences of Cladobotryum mycophilum (teleomorph Hypomyces odoratus) (GenBank Accession Nos. Y17096 and Y17095) (2). Pathogenicity tests were performed using 24 blocks containing sterilized, spawned, and incubated P. eryngii substrate (3.6 kg, 352 cm2 in area). The blocks were placed in a mushroom-growing room and cased with a 40-mm layer of a casing soil (0.7 liter block-1) made with mineral soil + Sphagnum peat 4:1 (vol/vol). Five days after casing, a conidial suspension (7 × 103 conidia ml-1) of one isolate of C. mycophilum was sprayed (5 ml per block) onto the surface of the casing layer at a rate of 106 conidia m-2. Twenty-two blocks were sprayed with sterile distilled water as a control. A temperature of 17 to 18°C and 85 to 90% relative humidity were maintained throughout cropping. The first cobweb symptoms developed 23 days after inoculation and C. mycophilum was consistently reisolated from nine (37.5%) of the inoculated blocks. Noninoculated blocks remained healthy. In a second test, conidial suspensions (3.4 × 105 conidia ml-1) of one isolate of C. mycophilum were inoculated onto 20 P. eryngii fruit bodies. Ten fruit bodies were inoculated externally while the other 10 fruit bodies were cut in half and inoculated internally with 50 μl of conidial suspension per fruit body. Sterilized distilled water was used as a control. All fruit bodies were then incubated at 22°C in a moist chamber. Assays were conducted twice and the results were recorded after 7 days. C. mycophilum grew on 85% of the internally inoculated fruit bodies and on 40% of those inoculated superficially, while the control mushrooms remained symptomless. To our knowledge, this is the first report of C. mycophilum causing cobweb in king oyster mushroom in Spain. This finding will have a potentially significant impact on button mushroom farms where cobweb is one of the most common diseases.

Postinflammatory depigmentation: excellent results with suction blister grafting

Depigmentation over the face of young females is usually not acceptable from an Indian social perspective. We are reporting a case of postinflammatory depigmentation over face of young female that was successfully treated. The patient developed depigmentation following inflammation as a result of a local application. She underwent three sessions of epidermal suction blister grafting with PUVA therapy instituted after the grafting to assist in the spreading of the pigmentation. First two sitting resulted in total repigmentation. She underwent a third sitting recently. Pigment spread is continuing with PUVA therapy in the area of the third session. The patient is satisfied with the outcome. In conclusion, we suggest that suction blister epidermal grafting can be tried in the treatment of postinflammatory depigmentation.

A Case of Piebaldism with Pigment Spread after Suction Blister Epidermal Grafting and PUVA Therapy

A Case of Piebaldism with Pigment Spread after Suction Blister Epidermal Grafting and PUVA Therapy

Renal transplant recipient infected with Penicillium marneffei

A 42-year-old woman presented to our hospital after 2 months of pain in her left hip. She had received a cadaveric-kidney transplant 9 months earlier at our hospital because of end-stage renal disease. After transplantation, the patient received maintenance immunosuppressants with oral tacrolimus (3 mg twice a day) and prednisolone (5 mg per day). On admission, she was afebrile. Her white-blood-cell count was 9150 cells per μL, with neutrophils of 62%, lymphocytes of 31%, and monocytes of 5%. Radiography showed an osteolytic lesion on her left pelvic brim adjacent to the acetabulum (fi gure). The osteolytic lesion was biopsied using an echoguided needle. Staining with gomori methenamine silver revealed yeast-like fungi with transverse septa (fi gure). At 25°C, this fungus grew as mold on sabouraud dextrose agar and produced a red pigment spreading into the agar (fi gure). On staining with lactophenol cotton blue, the conidiophores of this mold were smooth and had three to fi ve metulae, each of which with several phialides, producing smooth, spherical conidia in chains (fi gure). The mold-to-yeast conversion or phase transition, which is thermally regulated, is a diagnostic characteristic of Penicillium marneff ei. Intravenous liposomal amphotericin B (100 mg; 2 mg/kg body weight per day) was used for 21 days, followed by oral itraconazole (200 mg per day) for 8 months. No relapse of penicilliosis was seen 7 months after ending the antifungal treatment. P marneff ei has been reported to be an opportunistic infection endemic in areas of southeast Asia, including Thailand, southern China, Hong Kong, and Taiwan. Although infections in people with intact immune systems have been documented, the majority of reported cases involve patients with cellular immunity defects, receiving treatment with immunosuppressant drugs, or infected with HIV. Physicians should consider P marneff ei penicilliosis in patients that are immunocompromised who reside in or have a history of travelling to endemic areas.

Repigmentation of vitiligo with punch grafting and narrow‐band UV‐B (311 nm) – a prospective study

Phototherapy is already established as an effective mode of therapy in vitiligo. An evidence-based study was carried out of the effect of narrow-band (311 nm) ultraviolet-B (NB-UV-B) radiation in 66 surgically treated patients with recalcitrant vitiligo in whom autologous mini-punch grafting was deployed. A total of 2613 grafts were placed over 108 lesions on 17 regions in 66 individuals (39 females and 27 males) with stable, refractory vitiligo. The age range was 21-48 years. Postsurgically, they were exposed to a suberythemal dose of NB-UV-B (311 nm). Different parameters of surgical repigmentation were documented. Successful repigmentation was achieved in 57 (86.36%) cases. The appearance of repigmentation (AOR) time in different regions varied between 14 and 32 days, with an overall average of approximately 20.6 days. Maximum pigment spread (MPS) reached 12 mm with an average of 6.5 mm. The relationship between the donor graft area and area of surgical repigmentation was also calculated. Cobblestoning was the most common (31.8%) complication, but improved with time and/or interference. Punch grafting in combination with phototherapy (NB-UV-B, 311 nm) was found to be an easy, safe, inexpensive, and effective method of repigmenting static and stubborn vitiligo. Different facets of punch grafting-induced and phototherapy-aided surgical repigmentation were taken into consideration. The area of repigmentation, MPS, and relationship between the donor graft area and area of surgical repigmentation were documented.

Improving lip wrinkles: lipstick‐related image analysis

The appearance of lip wrinkles is problematic if it is adversely influenced by lipstick make-up causing incomplete color tone, spread phenomenon and pigment remnants. It is mandatory to develop an objective assessment method for lip wrinkle status by which the potential of wrinkle-improving products to lips can be screened. The present study is aimed at finding out the useful parameters from the image analysis of lip wrinkles that is affected by lipstick application. The digital photograph image of lips before and after lipstick application was assessed from 20 female volunteers. Color tone was measured by Hue, Saturation and Intensity parameters, and time-related pigment spread was calculated by the area over vermilion border by image-analysis software (Image-Pro). The efficacy of wrinkle-improving lipstick containing asiaticoside was evaluated from 50 women by using subjective and objective methods including image analysis in a double-blind placebo-controlled fashion. The color tone and spread phenomenon after lipstick make-up were remarkably affected by lip wrinkles. The level of standard deviation by saturation value of image-analysis software was revealed as a good parameter for lip wrinkles. By using the lipstick containing asiaticoside for 8 weeks, the change of visual grading scores and replica analysis indicated the wrinkle-improving effect. As the depth and number of wrinkles were reduced, the lipstick make-up appearance by image analysis also improved significantly. The lip wrinkle pattern together with lipstick make-up can be evaluated by the image-analysis system in addition to traditional assessment methods. Thus, this evaluation system is expected to test the efficacy of wrinkle-reducing lipstick that was not described in previous dermatologic clinical studies.