4 Skin eruptions are characterized by inflammatory, pruritic papules at infested sites; and the pathognomonic linear-to-serpiginous intradermal burrows. 4 Scabietic burrows are 5 to 10 mm long, dotted with fecal lithes (pellets) or scybala, and terminate in raised papules that may rarely hide ovipositing females. 4 Non-specific secondary lesions occur commonly from scratching (self-inflicted excoriations and lichenification); secondary infection (impetigo); or side effects of topical treatments (eczematization). Ribociclib The diagnosis of

scabies is made predominantly by epidemiological considerations and clinical and microscopic observations. A clinical diagnosis may be confirmed by low power microscopic examination of a burrow skin scraping which may excavate a female mite, 0.2 to 0.5 mm in length, translucent

with brown legs, and too small to be seen without magnification. 4 Eggs (0.02–0.03 mm in diameter), smaller eggshell fragments, and fecal lithes may also be identified in microscopic specimens of burrow scrapings. 4 Newer diagnostic methods for scabies now under investigation include enhanced microscopy (epiluminescence microscopy and non-computed dermoscopy); immunological detection of specific scabies antibodies by enzyme-linked immunosorbent assay (ELISA); and molecular identification of scabies mite DNA by polymerase chain reaction (PCR). 4,7,8 Topical or oral scabicides selleck chemical should be used to treat all infested persons and their close personal contacts simultaneously, regardless of the presence of symptoms. 4 Currently, recommended treatment options for scabies are listed in Table 2. The most effective topical treatments for scabies are 5% permethrin cream, 10 to 25% benzoyl benzoate lotion, and 1% lindane cream or lotion. 9 The topical treatments for scabies may not be well accepted or tolerated by some patients for many reasons including severe burning and stinging (with benzyl benzoate and 5% permethrin) in cases of secondarily excoriated or eczematous infestations. In such cases, a single oral dose of ivermectin, BCKDHB 200 mcg/kg, may

offer a more acceptable and equally effective alternative. 10 Nevertheless, ivermectin is not ovicidal, and a second course of oral treatment at adult maturation time of 14 to 15 days is recommended. 10 Scabies and follicle mites are the only exclusively human ectoparasitic mites and do not transmit infectious diseases. Less serious than scabies are infestations with the two human follicle mites: (1) Demodex folliculorum, which inhabits hair follicles; and (2) Demodex brevis, which inhabits sebaceous glands. The follicle mites are diminutive (0.1–0.4 mm long), usually non-pathogenic saprophytes, that feed on sebum and exfoliated skin while lodged in hair follicles and sebaceous glands on the eyelids, nasolabial folds, nose, and ears. 1 Although controversial, follicle mites may also be pathogenic agents of chronic blepharitis.

Too soon thereafter the announcement came along with a light tap on the shoulder, “Would a physician please identify himself or herself?” I jumped up and discovered to my disbelief that with roughly 300 passengers on this B-777, apparently I was the only physician. I was quickly ushered to the passenger, who was in the lavatory sitting on the toilet being held by her daughter. Much to my surprise, the patient happened not to be the transplant candidate, but the elderly woman who had looked like she harbored tuberculosis.

I squeezed onto the lavatory floor and obtained the history. She was 93, had a history of hypertension on metoprolol, and her daughter was taking her from her home in the Philippines to South Carolina as she could no longer care for herself. They had already just completed travel from the Philippines to JQ1 nmr Tokyo (several hours of ground travel followed by a 4-hour flight) and apparently her daughter thought her mother might feel better if she sat on the toilet and tried to relieve herself. I did not think this was going to be fruitful as the daughter also shared that her mother had not eaten much or drank since the onset of Inhibitor Library solubility dmso the trip. Unfortunately, the passenger (now patient) seemed to know just a few words in English and her daughter said that her

mother did not communicate very much anyway; this was not mitigated any by the fact that the daughter could not speak Filipino. I did not pursue details about their lack of ability to communicate with one another. My initial impression was that the patient may not have been too eager to leave her homeland for distant shores at this time in her life, and a candid discussion about this issue probably never occurred between them. Regardless, the patient could tell me only that she hurt all over. The enhanced

medical kit on many overseas flights is excellent (www.IATA.org/medical-manual), but for ideal use requires a team of health care providers and a bit more space than the typically oversold cabin. I found the blood pressure cuff and stethoscope to be useful. The patient’s blood pressure was 120/80, her pulse was regular, and I could not detect anything unusual on a cursory exam, except that she appeared somewhat cachectic and dry. She was not ADP ribosylation factor febrile. I could almost circle the largest part of her arm with my thumb and forefinger and her skin tented easily. She winced when I pressed anywhere, whether on her abdomen, chest, or limbs. She had no evidence of calf swelling, and moved all extremities equally. However, it was the sadness in her eyes that stayed with me. To start an intravenous line just for hydration would be difficult; her arm veins were tiny and collapsed. She would have required a neck or subclavian line that I was unprepared to place both because of the surroundings, but primarily due to my lack of expertise after so many years. I also doubted that this would have been her or her daughter’s choice at the moment.

Too soon thereafter the announcement came along with a light tap on the shoulder, “Would a physician please identify himself or herself?” I jumped up and discovered to my disbelief that with roughly 300 passengers on this B-777, apparently I was the only physician. I was quickly ushered to the passenger, who was in the lavatory sitting on the toilet being held by her daughter. Much to my surprise, the patient happened not to be the transplant candidate, but the elderly woman who had looked like she harbored tuberculosis.

I squeezed onto the lavatory floor and obtained the history. She was 93, had a history of hypertension on metoprolol, and her daughter was taking her from her home in the Philippines to South Carolina as she could no longer care for herself. They had already just completed travel from the Philippines to NVP-BGJ398 Tokyo (several hours of ground travel followed by a 4-hour flight) and apparently her daughter thought her mother might feel better if she sat on the toilet and tried to relieve herself. I did not think this was going to be fruitful as the daughter also shared that her mother had not eaten much or drank since the onset of EPZ-6438 concentration the trip. Unfortunately, the passenger (now patient) seemed to know just a few words in English and her daughter said that her

mother did not communicate very much anyway; this was not mitigated any by the fact that the daughter could not speak Filipino. I did not pursue details about their lack of ability to communicate with one another. My initial impression was that the patient may not have been too eager to leave her homeland for distant shores at this time in her life, and a candid discussion about this issue probably never occurred between them. Regardless, the patient could tell me only that she hurt all over. The enhanced

medical kit on many overseas flights is excellent (www.IATA.org/medical-manual), but for ideal use requires a team of health care providers and a bit more space than the typically oversold cabin. I found the blood pressure cuff and stethoscope to be useful. The patient’s blood pressure was 120/80, her pulse was regular, and I could not detect anything unusual on a cursory exam, except that she appeared somewhat cachectic and dry. She was not Fossariinae febrile. I could almost circle the largest part of her arm with my thumb and forefinger and her skin tented easily. She winced when I pressed anywhere, whether on her abdomen, chest, or limbs. She had no evidence of calf swelling, and moved all extremities equally. However, it was the sadness in her eyes that stayed with me. To start an intravenous line just for hydration would be difficult; her arm veins were tiny and collapsed. She would have required a neck or subclavian line that I was unprepared to place both because of the surroundings, but primarily due to my lack of expertise after so many years. I also doubted that this would have been her or her daughter’s choice at the moment.

We have shown previously that type IV pili (TFP) are required for wild-type levels of virulence of A. citrulli on melon and that this pathogen can colonize and move thorough the xylem vessels of host seedlings.

Here, comparative studies between wild-type and TFP mutant strains using microfluidic flow chambers demonstrated that TFP play a critical role in both the surface attachment and the biofilm formation of A. citrulli under a medium flow. Additionally, TFP null mutants were unable to perform twitching movement against the direction of medium Paclitaxel concentration flow. Assays using a flagellin mutant showed that, in contrast to TFP, polar flagella do not contribute to the adhesion and biofilm formation of A. citrulli under tested conditions. Also, flagellum-mediated swimming motility of wild-type strains was not observed under medium flow. These results imply that TFP may play an important role in colonization and spread in the xylem vessels under sap flow conditions, while polar flagella could be more important for spread during periods of time when xylem flow is minimal. Acidovorax avenae ssp. citrulli (Schaad et al., 1978; Willems et al., 1992) is a Gram-negative bacterium

that causes bacterial fruit blotch (BFB) of cucurbit plants. The destructive potential of this bacterium was fully realized during the late 1980s, following severe BFB outbreaks in watermelon fields in the United States that led to high yield

check details losses of up to 100% (Latin & Hopkins, 1995; Schaad et al., 2003). Since then, the Atazanavir disease has spread to different parts of the world, causing severe yield losses in watermelon and melon (Bahar & Burdman, 2010). Recently, Schaad et al. (2008) suggested a new classification for subspecies of A. avenae, with A. avenae ssp. citrulli being reclassified as Acidovorax citrulli. Herein, we adopt this new nomenclature. Molecular, biochemical and host-range characterization of A. citrulli isolates revealed the existence of two distinct groups: group I includes strains isolated mainly from nonwatermelon plants, while group II includes strains isolated mostly from watermelon (Walcott et al., 2000, 2004; Burdman et al., 2005; Feng et al., 2009). The genome of a group II strain (AAC00-1) has been sequenced recently by the Joint Genome Institute. Few studies have shed light on the transmission mechanisms of A. citrulli inside the plant. It has been shown that this bacterium can penetrate the plant through blossoms and subsequently infect seeds (Walcott et al., 2003; Lessl et al., 2007). In a recent study, we showed that A. citrulli can systemically infect melon seedlings and can move basipetally and acropetally through the xylem vessels (Bahar et al., 2009). The aforementioned studies suggest that motility contributes to both infection and translocation of the bacterium throughout the plant.

For example, it was reported that Caldicoprobacter oshimai was a xylanolytic, extremely thermophilic bacterium (Yokoyama et al., 2010). The OTU which showed the closest similarity with C. oshimai might also be a selleck chemical xylanolytic bacterium, which would play an important role in lignocellulose degradation. Another example was one OTU which represented 4% of the clone library, and one strain from the OTU which had been isolated

as pure culture. This strain was named ASX2 and shared 90.4% 16S rRNA gene sequence identity with Desulfotomaculum halophilum SEBR 3139. ASX2 was able to hydrolyze CMC, as determined by formation of a clear zone on a Congo Red agar plate (data not shown). Beta-glucosidase was also found in strain ASX2. It is noteworthy that most of the clones represented by the clone library HM781-36B shared 16S rRNA similarities lower than 90%, and all of them shared 16S rRNA similarities below 94%, which meant that they were novel at least at species level. One example was the isolation of strain ASX2 mentioned above. Another example was the isolated pure culture which shared 93% 16S rRNA sequence identity with Bacillus thermolactis. The general low 16S rRNA similarity might be attributable to the fact that the coastal marine environment was thought to be hypothermal, so thermophilic

bacteria were ignored. In our experiment, the selection pressure put on by thermophilic and anaerobic conditions and the limited carbon source eliminated bacterial species which were commonly found by traditional isolation methods under low temperature and aerobic conditions. The blast results

showed that the known strains most closely related to the sequenced clones were all from a terrigenous environment, for example Planifilum yunnanense isolated from a hot spring, Sporosalibacterium faouarense isolated from oil-contaminated www.selleck.co.jp/products/cobimetinib-gdc-0973-rg7420.html soil, D. halophilum isolated from an oilfield brine and C. oshimai isolated from sheep feces (Tardy-Jacquenod et al., 1998; Zhang et al., 2007; Yokoyama et al., 2010; Rezgui et al., 2011). A few of them such as D. halophilum and S. faouarense were reported as moderately halophilic bacteria (Tardy-Jacquenod et al., 1998; Rezgui et al., 2011). As we know, the marine environment was of high salinity and there is a possibility that these bacteria from land had adapted to the marine saline conditions and at last settled down in the ocean. However, the isolation environment and the low 16S rRNA similarity might indicate the opposite possibility, which was that the evolution positions of our clones pre-dated the isolated terrigenous strains. Briefly, the halophilic and thermophilic properties of these bacteria are unexpected and provide an interesting area for future studies. The phylogenetic tree of these clones and their closest related strains from the GenBank were constructed through the NJ method (Fig. 3). As shown in Fig. 3, all of the clones formed separate branches from the known species.

Δβ2tub (β2tub deletion) mutants were highly sensitive to MBC, produced fewer conidia and were less virulent than parental strains. Complementation of the Δβ2tub

mutants with a copy of the whole β2tub locus from their parental strains restored the level of MBC resistance (or sensitivity) to that of the parental strain. “
“Rhynchophorus ferrugineus is considered the worst pest of palm species, and few natural enemies are reported for this parasite in its area of origin. Here, we report the first recovery of the entomopathogenic fungus Metarhizium pingshaense associated with R. ferrugineus from Vietnam. The Dabrafenib datasheet morphological, biochemical, and toxicological features of this strain were studied and compared with those of another Metarhizium strain associated with this weevil in Sicily (Italy), an area of recent introduction. The potential use of these fungi as biocontrol agents was tested against adult insects in laboratory trials and a similar mortality rate was found. Both strains were able to produce toxins and cuticle-degrading proteases, but they showed dissimilar enzymatic and toxicological profiles, suggesting a different virulence activity. selleck chemicals “
“Bacterial swarming motility is a flagella-dependent

translocation on the surface environment. It has received extensive attention as a population behavior involving numerous genes. Here, we report that Citrobacter freundii, an opportunistic pathogen, exhibits swarming movement on a solid medium surface with appropriate agar concentration. The swarming behavior of C. freundii was described in detail. Insertional mutagenesis with transposon Mini-Tn5 was carried out to discover genetic determinants related to the swarming of C. freundii. A number of swarming genes were identified, among which flhD, motA, motB, wzx, rfaL, rfaJ, rfbX, rfaG, rcsD, rcsC, gshB, fabF, dam, pgi, and rssB have been characterized previously in other

species. In mutants related to lipopolysaccharide synthesis and RcsCDB signal system, a propensity to form poorly motile bacterial aggregates on the agar surface was observed. The aggregates hampered bacterial surface migration. In several PRKD3 mutants, the insertion sites were identified to be in the ORF of yqhC, yeeZ, CKO_03941, glgC, and ttrA, which have never been shown to be involved in swarming. Our results revealed several novel characteristics of swarming motility in C. freundii which are worthy of further study. Bacterial swarming is a flagella-dependent surface translocation exhibited by a wide variety of flagellated bacteria (for a review, see Allison & Hughes, 1991; Fraser & Hughes, 1999; Harshey, 2003; Kaiser, 2007; Kearns, 2010).

After nine days of inpatient admission (comparable to our usual average of three days), the family was

discharged home. When seen one week later on the outpatient clinic, the parents were coping better with the diagnosis. The diagnosis of type 1 diabetes in children places a huge psychological and emotional burden on the family. The diabetes-related stress of this mother can be associated with psychological distress and family conflict.1 Discomfort, 3-MA price anxiety, depression and post-traumatic stress symptoms can occur in mothers of children with type 1 diabetes.2,3 Interfering with traditional feeding patterns and activities can cause a lot of stress and family conflict. Needle fear and catastrophising pain by both patients and parents remain a major dilemma in the field of paediatric diabetes.4 Socio-demographic considerations

play a major role in the delivery MG-132 clinical trial for care of type 1 diabetes in youth.5 Culturally dictated lifestyles of the family may determine response to ‘scientific’ recommendations. It is well known that the delivery of diabetes care can be more efficient in ethnic minority groups when culturally competent interventions are utilised.6 Many studies have shown that the immigrant status of children can be a risk factor in the timing and diagnosis of type 1 diabetes in children7 as well as the progress of the development of complications of the disease.8 Understanding cultural, educational and economic factors of Amino acid immigrant and ethnic minority children with type 1 diabetes in any society is very crucial to improve their metabolic outcome.9 Proper diabetes education programmes and materials should be used when dealing with any family from an ethnic minority group with type 1 diabetes children. Respect for language barriers, strong different cultures

and health beliefs should be always exercised.10 Finally, the religious beliefs of the family should be respected as long as they do not seem to pose an imminent danger to the life of the child with type 1 diabetes. There are a few case reports of children with type 1 diabetes who have died at home while parents did not perform expected therapy because of their religious beliefs. Health care providers need to be observant of warning signs of dangerous beliefs that may result in the death of a diabetic child while parents are praying for a cure.11 Ethnic minorities and immigrant families may pose some challenges when it comes to dealing with type 1 diabetes in their children. Cultural differences, religious beliefs and unreasonable expectations from their new western society may interfere with the delivery of diabetes education and prolonged hospital stay. Fear, mistrust, and financial and social stresses should be also addressed.

Indeed, a representative species, Prevotella intermedia, is recognized as one of the major periodontal pathogens (Socransky & Haffajee, 2005). Prevotella intermedia possesses various virulence

factors such as adhesin, hemolysin, and hemagglutinin, as well as proteolytic and hydrolytic enzymes, which allows it to colonize the oral cavity, evade host defenses, modulate immune responses, and cause tissue destruction (Eley & Cox, 2003). Indole, which has been used for decades as a taxonomic tool for the identification of gram-negative rods (Hütter & DeMoss, 1967; Wolfe & Amsterdam, 1968), is one of a host of malodorous oral volatile products that also includes methyl mercaptan, hydrogen sulfide, skatole, and cadaverine (Fosdick

& Piez, 1953; Kostelc et al., 1981; Claesson et al., 1990; Goldberg et al., 1994). Interestingly, indole has recently selleck compound been reported to regulate bacterial Ku-0059436 cost biofilms (Lee et al., 2007; Sasaki-Imamura et al., 2010), multidrug exporters (Hirakawa et al., 2005), and a pathogenicity island (Anyanful et al., 2005). It was reported more than a half century ago that the saliva of patients with periodontal disease contains high levels of indole (Berg et al., 1946). In addition, certain periodontopathogenic bacteria, including P. intermedia, Porphyromonas gingivalis, and Fusobacterium nucleatum, have the capacity to produce indole from l-tryptophan (Duerden et al., 1980). These findings suggest that indole may play an important role in the onset and progression of periodontitis. Indole is produced via α,

β-elimination Flavopiridol (Alvocidib) of l-tryptophan in a reaction catalyzed by tryptophanase, which is encoded by tnaA (Snell, 1975). We recently identified and characterized tryptophanase from P. gingivalis W83 (Yoshida et al., 2009) and F. nucleatum ATCC 25586 (Sasaki-Imamura et al., 2010). The enzymatic properties of P. gingivalis tryptophanase were similar to those of F. nucleatum, but the bacteria differed in tnaA gene organization and in the manner in which the tnaA region was transcribed. In the current study, the tnaA gene encoding tryptophanase in P. intermedia ATCC 25611T was identified and sequenced. The transcriptional unit of the gene was investigated and purified recombinant tryptophanase was enzymatically characterized. The presence of tnaA and the capacity to produce indole were examined in several members of the genus Prevotella. Twenty-two strains of the genus Prevotella, P. gingivalis ATCC 33277, and F. nucleatum ssp. nucleatum ATCC 25586 were obtained from RIKEN BioResource Center (Wako, Japan). All Prevotella strains were isolated from oral and craniofacial regions (Table 1). All of the strains were grown anaerobically at 37 °C in enriched brain–heart infusion (BHI) broth (Yoshida et al., 2009).

The DSR system presumably forms intracellular sulfite that is oxidized by an enzyme system consisting of Sat, Apr, and Qmo proteins (Rodriguez et al., 2011). The electron acceptors, cytochrome c, and menaquinone (Fig. 1) are ultimately oxidized

by the photosynthetic reaction center. In cultures of Cba. tepidum that contain both sulfide and thiosulfate, sulfide is oxidized preferentially while sulfur globules are formed (Chan et al., 2008; Azai et al., 2009; Holkenbrink et al., 2011). Following sulfide depletion, thiosulfate and sulfur globules are oxidized to sulfate. The molecular mechanism of learn more this phenomenon is poorly understood. Sulfide possibly inhibits thiosulfate oxidation either by substrate competition between sulfide and thiosulfate (the SOX system oxidizes sulfide in vitro; Ogawa et al., 2010) or by saturation of the electron acceptor pool. Regulation of sulfur metabolism genes in GSB is poorly described, but it is known that SoxA is induced by thiosulfate in Chlorobaculum thiosulfatiphilum (Verté et al. 2002). DAPT in vitro In the purple sulfur bacterium, Allochromatium vinosum, sox and dsr genes are expressed at a low constitutive level in the absence of reduced sulfur substrates and are induced by thiosulfate and sulfide, respectively (Grimm et al., 2010, 2011). Chlorobaculum tepidum TLS was grown under incandescent illumination in CL medium (Frigaard et al., 2004). For experiments comparing early and late exponential growth phase, wild-type

cells were grown at 45 °C in 1-L flasks under a light intensity of 200 μmol photons m−2 s−1. For experiments comparing wild type and the dsrM mutant (Holkenbrink et al., 2011), cells were grown at 42 °C in 15-mL

tubes under a light intensity of 50 μmol photons m−2 s−1 and harvested in the late exponential growth phase. Cells were harvested by centrifugation and stored at −20 °C prior to analysis. Bacteriochlorophyll c was determined by extracting the cell pellet with acetone : methanol (7 : 2 by vol) (Frigaard et al., 1997). Sulfide was measured using the colorimetric methylene blue method (Cline 1969). Sulfate acetylcholine and thiosulfate were measured by ion chromatography (Dionex, Hvidovre, Denmark) using a carbonate buffer as eluent. Samples for analysis of elemental sulfur were dissolved in methanol and analyzed as S8 by HPLC using a Sykam pump (S1100), UV–VIS detector (Sykam S3200), Zorbax ODS-column (125 × 4 mm, 5 μm; Knauer, Berlin, Germany) and methanol as the eluent at a flow rate of 1 mL min−1. Elemental sulfur was detected at 265 nm. Cell pellets were thawed and extracted in acetone : methanol (7 : 2 by vol) to remove pigments. The colorless cell pellets were solubilized in an SDS-containing buffer (Laemmli, 1970) supplemented with a complete protease inhibitor cocktail (Roche, Hvidovre, Denmark) at 95 °C for 3–5 min and cleared by centrifugation. Prior to digestion, proteins were reduced with dithiothreitol (1 mM) and alkylated with iodoacetamide (5 mM).

The DSR system presumably forms intracellular sulfite that is oxidized by an enzyme system consisting of Sat, Apr, and Qmo proteins (Rodriguez et al., 2011). The electron acceptors, cytochrome c, and menaquinone (Fig. 1) are ultimately oxidized

by the photosynthetic reaction center. In cultures of Cba. tepidum that contain both sulfide and thiosulfate, sulfide is oxidized preferentially while sulfur globules are formed (Chan et al., 2008; Azai et al., 2009; Holkenbrink et al., 2011). Following sulfide depletion, thiosulfate and sulfur globules are oxidized to sulfate. The molecular mechanism of Pirfenidone in vivo this phenomenon is poorly understood. Sulfide possibly inhibits thiosulfate oxidation either by substrate competition between sulfide and thiosulfate (the SOX system oxidizes sulfide in vitro; Ogawa et al., 2010) or by saturation of the electron acceptor pool. Regulation of sulfur metabolism genes in GSB is poorly described, but it is known that SoxA is induced by thiosulfate in Chlorobaculum thiosulfatiphilum (Verté et al. 2002). Palbociclib price In the purple sulfur bacterium, Allochromatium vinosum, sox and dsr genes are expressed at a low constitutive level in the absence of reduced sulfur substrates and are induced by thiosulfate and sulfide, respectively (Grimm et al., 2010, 2011). Chlorobaculum tepidum TLS was grown under incandescent illumination in CL medium (Frigaard et al., 2004). For experiments comparing early and late exponential growth phase, wild-type

cells were grown at 45 °C in 1-L flasks under a light intensity of 200 μmol photons m−2 s−1. For experiments comparing wild type and the dsrM mutant (Holkenbrink et al., 2011), cells were grown at 42 °C in 15-mL

tubes under a light intensity of 50 μmol photons m−2 s−1 and harvested in the late exponential growth phase. Cells were harvested by centrifugation and stored at −20 °C prior to analysis. Bacteriochlorophyll c was determined by extracting the cell pellet with acetone : methanol (7 : 2 by vol) (Frigaard et al., 1997). Sulfide was measured using the colorimetric methylene blue method (Cline 1969). Sulfate Cediranib (AZD2171) and thiosulfate were measured by ion chromatography (Dionex, Hvidovre, Denmark) using a carbonate buffer as eluent. Samples for analysis of elemental sulfur were dissolved in methanol and analyzed as S8 by HPLC using a Sykam pump (S1100), UV–VIS detector (Sykam S3200), Zorbax ODS-column (125 × 4 mm, 5 μm; Knauer, Berlin, Germany) and methanol as the eluent at a flow rate of 1 mL min−1. Elemental sulfur was detected at 265 nm. Cell pellets were thawed and extracted in acetone : methanol (7 : 2 by vol) to remove pigments. The colorless cell pellets were solubilized in an SDS-containing buffer (Laemmli, 1970) supplemented with a complete protease inhibitor cocktail (Roche, Hvidovre, Denmark) at 95 °C for 3–5 min and cleared by centrifugation. Prior to digestion, proteins were reduced with dithiothreitol (1 mM) and alkylated with iodoacetamide (5 mM).