Spent Coffee Grounds and Novaluron Are Toxic to Aedes aegypti (Diptera: Culicidae) Larvae.

Aedes aegypti (Diptera: Culicidae) is a vector for mosquito-borne diseases worldwide. Insecticide resistance is a major concern in controlling this mosquito. We investigated the chemical compounds in wet and dry spent coffee grounds (wSCGs and dSCGs) and evaluated the efficacy of dSCGs, wSCGs, and novaluron on the mortality and adult emergence inhibition of Ae. aegypti. We found higher concentrations of chemical compounds in wSCGs than in dSCGs. The wSCGs and dSCGs both contained total phenolic compounds, total flavonoid compounds, caffeic acid, coumaric acid, protocatechuic acid, and vanillic acid. Complete mortality was observed after 48 h of exposure to 50 g/L wSCGs, while similar mortality was found after 120 h of exposure to 10 µg/L of novaluron. The sublethal dose was a concentration of wSCGs (5 g/L) and novaluron (0.01, 0.1, and 1 µg/L) combined that resulted in a larval mortality lower than twenty percent (at 72 h) to determine their synergistic effects. The death rate of larvae exposed in sublethal combination of wSCGs and novaluron was significantly higher than that of its stand-alone. The findings indicate that the combination of wSCGs and novaluron at sublethal concentrations had synergistic effects on the mortality of Ae. aegypti larvae and could be applied as an alternative control measure.

Species Identification of the Major Japanese Encephalitis Vectors within the Culex vishnui Subgroup (Diptera: Culicidae) in Thailand Using Geometric Morphometrics and DNA Barcoding.

Japanese encephalitis (JE) is a viral infection of the brain caused by the Japanese encephalitis virus, which spreads globally, particularly in 24 countries of Southeast Asia and the Western Pacific region. In Thailand, the primary vectors of JE are Cx. pseudovishnui, Cx. tritaeniorhynchus, and Cx. vishnui of the Cx. vishnui subgroup. The morphologies of three mosquito species are extremely similar, making identification challenging. Thus, geometric morphometrics (GM) and DNA barcoding were applied for species identification. The results of cross-validation reclassification revealed that the GM technique based on wing shape analysis had relatively high potential for distinguishing Cx. pseudovishnui, Cx. tritaeniorhynchus, and Cx. vishnui (total performance = 88.34% of correctly assigned individuals). While the DNA barcoding yielded excellent results in identifying these Culex species based on the DNA barcode gap (average intraspecific genetic distance = 0.78% ± 0.39% and average interspecific genetic distance = 6.14% ± 0.79%). However, in the absence of the required facilities for DNA barcoding, GM techniques can be employed in conjunction with morphological methods to enhance the reliability of species identification. Based on the results of this study, our approach can help guide efforts to identify members of the Cx. vishnui subgroup, which will be useful for the effective vector control of JE in Thailand.

Incobotulinum Toxin A with a One-year Long-lasting Effect for Trapezius Contouring and Superior Efficacy for the Treatment of Trapezius Myalgia.

Context: Based on various Botulinum toxin A products, reports of the lower efficacy of Incobotulinum toxin A compared with Onabotulinum toxin A for muscle contouring were observed. In addition, complications of trapezius myalgia and shoulder contouring treatment from malpractice have been reported. Aims: The study aimed at comparing the efficacy between Incobotulinum toxin A and Onabotulinum toxin A; research was conducted on a safe treatment technique for trapezius hypertrophy and trapezius myalgia. Materials and Methods: A split-shoulder, double-blind, randomized controlled trial was performed. Twenty volunteers with trapezius hypertrophy and trapezius myalgia were randomly injected with 30 units of Incobotulinum toxin A and Onabotulinum toxin A in each trapezius muscle guided by ultrasound. Results: The trapezius thickness among those receiving treatment with Onabotulinum toxin A and Incobotulinum toxin A on day 60 was 7.35 ± 1.11 and 7.33 ± 1.21 mm, respectively, which did not portray a significant difference (P = 0.991). Compared with the muscle size from day 60 to one year, the size of the trapezius muscle that had been treated by Onabotulinum toxin type A regained a significantly larger size compared with that treated by Incobotulinum toxin A (P = 0.027). On comparing the size of the trapezius muscle treated by Incobotulinum toxin A between one year and day 0, it was observed that the trapezius thickness at one year had significantly decreased (P < 0.001). On comparing the pain score from day 60 to day 0, it was observed that the pain scores of trapezius myalgia treated by Onabotulinum toxin A and Incobotulinum toxin A significantly differed (P = 0.003). Conclusions: Incobotulinum toxin A had the same efficacy but a longer lasting effect for the trapezius size contouring and a higher efficacy for trapezius myalgia treatment compared with Onabotulinum toxin A.

Adjunctive treatment for acne vulgaris by tranexamic acid.

BACKGROUND: Acne is a chronic, inflammatory skin disorder of pilosebaceous units. Tranexamic acid (TXA) acts as a plasmin inhibitor to reduce blood loss and is also used to treat rosacea due to its anti-inflammatory effects. Some parts of the pathogenesis of rosacea are similar to inflammatory acne. OBJECTIVES: The study aimed to evaluate the efficacy of 10% TXA serum in treating acne and its adverse effects. METHODS: A randomized, double-blind, placebo-controlled, split-face study was performed on 18 mild-to-moderate acne patients. Patients applied 10% TXA serum on one side of the face and placebo on another side twice daily for 8 weeks. Acne lesion counts and adverse effects were evaluated every 2 weeks. RESULTS: Significant differences in total inflammatory acne counts were observed between TXA and placebo since Week 4 (p = 0.008). TXA mainly reduced papules and pustules, as papule counts significantly decreased since the 8th week (p = 0.046) and pustule counts significantly reduced since Week 8 (p = 0.033). Moreover, physicians also found that TXA serum reduced the redness of the skin, corresponding with the imaging from VISIA® Skin Analysis. The anti-inflammatory effect of TXA resulted in less PIE and PIH. Adverse effects, including erythema and scaling, were treated by applying any moisturizing cream. CONCLUSION: Topical 10% TXA can reduce inflammatory acne effectively. Adverse effects were minor and treat easily.

Development of a novel multiplex PCR assay for the detection and differentiation of Plasmodium caprae from Theileria luwenshuni and Babesia spp. in goats.

Intraerythrocytic parasites are traditionally identified by the microscopic examination of Giemsa-stained blood smears. However, this method does not always allow for the identification of individual species in goat's RBCs. Moreover, its unreliability in detecting low levels of parasitemia makes it unsuitable for epidemiological investigations and leaves goat farms vulnerable to potential outbreaks. In the present study, a novel multiplex PCR (mPCR) targeting the cytochrome c oxidase subunit I (COI) gene was developed to detect and subsequently differentiate Plasmodium caprae, Theileria luwenshuni, and Babesia spp. The specificity of each primer set was assessed both in silico and with a panel of DNA samples from the hosts themselves and other goat hemoparasites. Amplicons generated from each pair of primers were 664, 555, and 320-bp for P. caprae, Babesia spp., and T. luwenshuni, respectively. These products were further confirmed by sequencing. Our novel mPCR reactions successfully demonstrated the accurate and simultaneous amplification of the three parasites' DNA samples. The current mPCR method showed no cross-amplification with unintended targets. The detection limit of the mPCR in this study was 108 parasites' DNA copies per reaction. The current mPCR was able to detect the minimum parasitemia of approximately 0.001%, 0.000005%, 0.00001% for P. caprae, Babesia spp. and T. luwenshuni, respectively. The diagnostic specificity in the detection of P. caprae and T. luwenshuni ranged from 94.9 to 100 %. The mPCR was further applied to a collection of field blood samples from five provinces in Thailand to validate its reliability and applicability. The results demonstrated the successful detection of P. caprae, Babesia spp. and T. luwenshuni in goat samples with the same sensitivity levels as conventional PCR methods. This study also confirmed the presence of T. luwenshuni and Babesia spp. in Thai goats. The current mPCR method offers an alternative for the diagnosis of P. caprae, T. luwenshuni, and Babesia spp., either single or under co-infection conditions, and for large-scale surveillance.

Development and application of a novel multiplex PCR assay for the differentiation of four haemosporidian parasites in the chicken Gallus gallus domesticus.

Haemosporidian infections in domestic chickens (Gallus gallus domesticus) are not only widely prevalent but also cause economic loss. Diagnosis is usually made by microscopic examination; however, the method has several drawbacks such as requiring an experienced microscopist, being unreliable when parasitemia is low and being unable to accurately differentiate between co-infections from multiple parasite species. Therefore, the current extent of haemosporidian infections might be underestimated and neglected. We have developed a novel multiplex PCR assay to simultaneously detect and differentiate between four haemosporidian parasites: Leucocytozoon caulleryi, Leucocytozoon sabrazesi, Plasmodium juxtanucleare and Plasmodium gallinaceum. Primers in the present study specifically amplified the corresponding targets with no cross-species amplification detected. The multiplex PCR exhibited a significantly greater detection rate when compared with microscopic examination (p = 0.0001). The results demonstrate that the detection rate of multiplex PCR for L. sabrazesi, P. juxtanucleare, and P. gallinaceum are all greater than that of microscopic examination with p = 0.002, 0.0001 and 0.004, respectively. Co-infections were also detected more effectively by multiplex PCR. We applied the current method to field samples originating from Nan, Prachinburi, and Chachoengsao Provinces. The current study revealed that positive rates of haemosporidian parasites in chickens in the three study sites ranging from 39.5%-93.8%. The present assay offers a timesaving option for molecular diagnosis instead of using singleplex PCRs for detecting the parasites individually. Within a single reaction, this assay would be a useful tool for the detection of avian haemosporidian parasites either single or under co-infection conditions and for large-scale epidemiology studies.

Synbiotics supplement is effective for Melasma improvement.

BACKGROUND: Melasma is a disorder of melanogenesis among humans causing localized, chronic acquired hypermelanosis of the skin requiring a combination of treatments. Related studies have shown probiotics contribute distinct advantages for skin disorders possibly including melasma because of its anti-inflammatory activities, anti-oxidation properties, ultraviolet protection, and tyrosinase activity inhibition. AIMS: The study aimed to investigate the effects of synbiotics supplement on improving melasma (evaluated from mMASI score). METHODS: This research comprised an experimental study employing a prospective, double-blind, randomized controlled trial among 57 Thai participants divided in 2 groups (29 for the experimental and 28 for the placebo groups). The participants were aged 30-50 years old, had Fitzpatrick skin type III-VI, with facial melasma on both sides of the face and attending Mae Fah Luang University Hospital, Bangkok from January-December 2019. Participants were randomly treated with oral synbiotics or placebo, 1 sachet daily for 12 weeks. Melasma severity and skin health were evaluated at 4 visits for each participant (baseline, weeks 4, 8, and 12, respectively). RESULTS: Severity of melasma scored by mMASI of the synbiotics group was 7.54 ± 0.79, 7.36 ± 0.80, 7.16 ± 0.73, and 6.98 ± 0.72 at baseline, weeks 4, 8, and 12, respectively, and 7.51 ± 0.86, 7.52 ± 0.88, 7.54 ± 0.86, and 7.54 ± 0.89 at baseline, weeks 4, 8, and 12, respectively, in the placebo group. Comparing between two groups at week 12, melasma score in the synbiotics supplement group was significantly lower than that in the placebo group (P = .008). CONCLUSION: Oral synbiotics supplementation for 12 weeks improved the severity of melasma score.

Author Correction: Genetic homogeneity of goat malaria parasites in Asia and Africa suggests their expansion with domestic goat host.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.

Genetic homogeneity of goat malaria parasites in Asia and Africa suggests their expansion with domestic goat host.

Plasmodium was first identified in a goat in Angola in 1923, and only recently characterized by DNA isolation from a goat blood sample in Zambia. Goats were first domesticated in the Fertile Crescent approximately 10,000 years ago, and are now globally distributed. It is not known if the Plasmodium identified in African goats originated from parasites circulating in the local ungulates, or if it co-evolved in the goat before its domestication. To address this question, we performed PCR-based surveillance using a total of 1,299 goat blood samples collected from Sudan and Kenya in Africa, Iran in west Asia, and Myanmar and Thailand in southeast Asia. Plasmodium DNA was detected from all locations, suggesting that the parasite is not limited to Africa, but widely distributed. Whole mitochondrial DNA sequences revealed that there was only one nucleotide substitution between Zambian/Kenyan samples and others, supporting the existence of a goat-specific Plasmodium species, presumably Plasmodium caprae, rather than infection of goats by local ungulate malaria parasites. We also present the first photographic images of P. caprae, from one Kenyan goat sample.

Artesunate-tafenoquine combination therapy promotes clearance and abrogates transmission of the avian malaria parasite Plasmodium gallinaceum.

Clinical manifestations of malaria infection in vertebrate hosts arise from the multiplication of the asexual stage parasites in the blood, while the gametocytes are responsible for the transmission of the disease. Antimalarial drugs that target the blood stage parasites and transmissible gametocytes are rare, but are essentially needed for the effective control of malaria and for limiting the spread of resistance. Artemisinin and its derivatives are the current first-line antimalarials that are effective against the blood stage parasites and gametocytes, but resistance to artemisinin has now emerged and spread in various malaria endemic areas. Therefore, a novel antimalarial drug, or a new drug combination, is critically needed to overcome this problem. The objectives of this study were to evaluate the efficacy of a relatively new antimalarial compound, tafenoquine (TQ), and a combination of TQ and a low dose of artesunate (ATN) on the in vivo blood stage multiplication, gametocyte development and transmission of the avian malaria parasite Plasmodium gallinaceum to the vector Aedes aegypti. The results showed that a 5-d treatment with TQ alone was unable to clear the blood stage parasites, but was capable of reducing the mortality rate, while TQ monotherapy at a high dose of 30mg/kg was highly effective against the gametocytes and completely blocked the transmission of P. gallinaceum. In addition, the combination therapy of TQ+ATN completely cleared P. gallinaceum blood stages and sped up the gametocyte clearance from chickens, suggesting the synergistic effect of the two drugs. In conclusion, TQ is demonstrated to be effective for limiting avian malaria transmission and may be used in combination with a low dose of ATN for safe and effective treatment.

Molecular detection of the avian malaria parasite Plasmodium gallinaceum in Thailand.

Avian malaria is one of the most common veterinary problems in Southeast Asia. The standard molecular method for detection of the avian malaria parasite involves the phenol-chloroform extraction of parasite genomic (g)DNA followed by the amplification of parasite gDNA using polymerase chain reaction (PCR). However, the phenol-chloroform extraction method is time-consuming and requires large amounts of samples and toxic organic solvents, thereby limiting its applications for parasite detection in the field. This study aimed to compare the performance of chelex-100 resin and phenol/chloroform extraction methods for the extraction of Plasmodium gallinaceum gDNA from whole avian blood that had been dried on filter papers (a common field sampling method). The specificity and sensitivity of PCR assays for P. gallinaceum cytochrome B (cytb) and cytochrome oxidase subunit I (coxI) gene fragments (544 and 588bp, respectively) were determined, and found to be more sensitive with gDNA extracted by the chelex-100 resin method than with the phenol/chloroform method. These PCR assays were also performed to detect P. gallinaceum in 29 blood samples dried on filter papers from domestic chickens in a malaria endemic area, where the reliable identification of seven field isolates of P. gallinaceum was obtained with an accuracy of 100%. The analysis of cytb and coxI gene nucleotide sequences revealed the existence of at least two genetically distinct populations of P. gallinaceum in Thailand, both of which differed from the reference strain 8A of P. gallinaceum. In conclusion, the chelex-100 resin extraction method is a simple and sensitive method for isolating gDNA from whole avian blood dried on filter paper. Genomic DNA extracted by the chelex method could subsequently be applied for the PCR-based detection of P. gallinaceum and DNA sequencing. Our PCR assays provide a reliable diagnostic tool for molecular epidemiological studies of P. gallinaceum infections in domestic chickens and wild birds.

Effects of artesunate treatment on Plasmodium gallinaceum transmission in the vectors Aedes aegypti and Culex quinquefasciatus.

In the absence of vaccines, chemotherapy is an effective and economical way for controlling malaria. Development of anti-malarial drugs that target pathogenic blood stage parasites and gametocytes is preferable for the treatment as it can alleviate the host's morbidity and mortality and block transmission of the Plasmodium parasite. Recently, our laboratory has developed an in vivo transmission blocking assay that involves administration of 7 consecutive daily doses of a test compound into domestic chickens (Gallus gallus domesticus) infected with the avian malaria parasite Plasmodium gallinaceum with 10% parasitaemia and 1% gametocytaemia. To compromise the cost and time for artesunate (ATN) treatment, this study aimed to investigate effects of a 5-day consecutive administration of 10 milligrams per kilogram (mg/kg) ATN on P. gallinaceum infection in chickens and transmission to two natural vectors, Aedes aegypti and Culex quinquefasciatus. Our study showed that the treatment with 10 mg/kg ATN for 7 days, but not 5 days, completely eliminated blood stage infections, prevented recrudescence and blocked gametocyte production and transmission of P. gallinaceum to its vectors, thereby confirming the potent schizontocidal and gametocytocidal activities of ATN. This regimen should be further evaluated in field trials.

In vivo transmission blocking activities of artesunate on the avian malaria parasite Plasmodium gallinaceum.

Infection and transmission of the avian malaria parasite Plasmodium gallinaceum in domestic chickens is associated with high economic burden and presents a major challenge to poultry industry in South East Asia. Development of drugs targeting both asexual blood stage parasites and sexual stages of the avian malarias will be beneficial for malaria treatment and eradication. However, current drugs recommended for treatment of the avian malaria parasites target specifically the asexual blood stage parasites, but have little or no impact to the gametocytes, the major target for development of transmission-blocking strategies. In the present work, we established a simple procedure to evaluate gametocytocidal and transmission blocking activities in a P. gallinaceum-avian model. The assays involved administration of seven consecutive daily doses of test compounds into P. gallinaceum-infected chickens with 10% parasitaemia and 1% gametocytaemia. Our studies indicated that intramuscular injection with seven daily low doses (the minimum effective dose of 10mg/kg) of artesunate blocked the gametocyte production and transmission to the mosquito vector Aedes aegypti. This assay can be further applicable for testing new compounds against P. gallinaceum and for other parasitic protozoa infecting birds.

Green tea extract supplement inhibition of HMGB1 release in rats exposed to cigarette smoke.

Tobacco-smoke exposure is linked to carcinogenic, oxidative and inflammatory cellular reactions. Green tea has been reported to have anti-release properties against various pro-inflammatory cytokines. To determine the effects of green tea extract (GTE) on serum high mobility group box-1 (HMGB1) levels in rats exposed to cigarette smoke (CS), we divided rats into 4 treatment groups: (1) CS only, (2) dietary supplement with GTE (3 mg/d) and CS (GCS1), (3) dietary supplement with GTE (4.5 mg/d) and CS (GCS2) and (4) a control group. HMGB1 and cotinine serum levels were analyzed by ELISA. The average serum HMGB1 level in the CS group was significantly higher than the other groups (p < 0.01), indicating the release of HMGB1 into the blood was stimulated by CS exposure, while GTE consumption suppressed HMGB1 levels. Rats exposed to CS had an average serum cotinine level of 37 ng/ml, indicating tobacco related compounds were present in the rats' blood. However, treatment with GTE did not reduce cotinine levels in all groups. Cotinine stimulated HMGB1 secretion in a dose- and time-dependent manner, and HMGB1 levels were suppressed by GTE in murine macrophage cell lines. Our results show GTE supplementation may offer beneficial systemic effects and suppress HMGB1 by protecting against cell inflammation.