A novel homozygous frameshift variant in DNAH8 causes multiple morphological abnormalities of the sperm flagella in a consanguineous Pakistani family / 亚洲男科学杂志(英文版)

Multiple morphological abnormalities of the sperm flagella (MMAF) is a severe form of asthenozoospermia categorized by immotile spermatozoa with abnormal flagella in ejaculate. Whole-exome sequencing (WES) is used to detect pathogenic variants in patients with MMAF. In this study, a novel homozygous frameshift variant (c.6158_6159insT) in dynein axonemal heavy chain 8 (DNAH8) from two infertile brothers with MMAF in a consanguineous Pakistani family was identified by WES. Reverse transcription-polymerase chain reaction (RT-PCR) confirmed DNAH8 mRNA decay in these patients with the DNAH8 mutation. Hematoxylin-eosin staining and transmission electron microscopy revealed highly divergent morphology and ultrastructure of sperm flagella in these patients. Furthermore, an immunofluorescence assay showed the absence of DNAH8 and a reduction in its associated protein DNAH17 in the patients' spermatozoa. Collectively, our study expands the phenotypic spectrum of patients with DNAH8-related MMAF worldwide.

Novel biallelic loss-of-function mutations in / 亚洲男科学杂志(英文版)

Multiple morphological abnormalities of the sperm flagella (MMAF) is a specific type of asthenoteratozoospermia, presenting with multiple morphological anomalies in spermatozoa, such as absent, bent, coiled, short, or irregular caliber flagella. Previous genetic studies revealed pathogenic mutations in genes encoding cilia and flagella-associated proteins (CFAPs; e.g., CFAP43, CFAP44, CFAP65, CFAP69, CFAP70, and CFAP251) responsible for the MMAF phenotype in infertile men from different ethnic groups. However, none of them have been identified in infertile Pakistani males with MMAF. In the current study, two Pakistani families with MMAF patients were recruited. Whole-exome sequencing (WES) of patients and their parents was performed. WES analysis reflected novel biallelic loss-of-function mutations in CFAP43 in both families (Family 1: ENST00000357060.3, p.Arg300Lysfs*22 and p.Thr526Serfs*43 in a compound heterozygous state; Family 2: ENST00000357060.3, p.Thr526Serfs*43 in a homozygous state). Sanger sequencing further confirmed that these mutations were segregated recessively in the families with the MMAF phenotype. Semiquantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) was carried out to detect the effect of the mutation on mRNA of the affected gene. Previous research demonstrated that biallelic loss-of-function mutations in CFAP43 accounted for the majority of all CFAP43-mutant MMAF patients. To the best of our knowledge, this is the first study to report CFAP43 biallelic loss-of-function mutations in a Pakistani population with the MMAF phenotype. This study will help researchers and clinicians to understand the genetic etiology of MMAF better.

Insight on multiple morphological abnormalities of sperm flagella in male infertility: What is new? / 亚洲男科学杂志(英文版)

The syndrome of multiple morphological abnormalities of the sperm flagella (MMAF) is a specific kind of asthenoteratozoospermia with a mosaic of flagellar morphological abnormalities (absent, short, bent, coiled, and irregular flagella). MMAF was proposed in 2014 and has attracted increasing attention; however, it has not been clearly understood. In this review, we elucidate the definition of MMAF from a systematical view, the difference between MMAF and other conditions with asthenoteratozoospermia or asthenozoospermia (such as primary mitochondrial sheath defects and primary ciliary dyskinesia), the knowledge regarding its etiological mechanism and related genetic findings, and the clinical significance of MMAF for intracytoplasmic sperm injection and genetic counseling. This review provides the basic knowledge for MMAF and puts forward some suggestions for further investigations.

A probable hybrid female Psychodopygus sp. (Diptera, Psychodidae, Phlebotominae) specimen from Amazonas State, Brazil / Un híbrido probable en una hembra Psychodopygus sp. (Diptera, Psychodidae, Phlebotominae) en el estado de Amazonas, Brasil

Phlebotomine sand flies are vectors of several important etiological agents. Sand fly cibaria and genitalia exhibit morphological characters that are essential for species identification. A morphologically anomalous female sand fly of the subgenus Psychodopygus Mangabeira was found in a faunistic survey of a forested area near Manaus, Amazonas State, Brazil. This specimen possesses two pairs of spermathecae and a cibarium with supernumerary rows of teeth. Most morphological anomalies in sand flies occur in the number and arrangement of spines in structures associated with the male genitalia. In females, the number of cibarial teeth is the most common form of anomaly. The specimen described here constitutes a rare anomalous record.

Los flebótomos son importantes vectores de varios agentes etiológicos. En flebótomos, los caracteres morfológicos del cibario y la genitália, son esenciales para la correcta identificación de las especies. Una hembra morfológicamente anómala del subgénero Psychodopygus Mangabeira, fue encontrada en un levantamiento faunístico en un área de bosque cerca de Manaus, estado de Amazonas, Brasil. Este espécime tiene dos pares de espermatecas y un cibario como hileras de dientes supranumerários. La mayoria de las anomalías morfológicas en los flebótomos ocurre en el número y disposición de las espinas en las estruturas de la genitália masculina. En las hembras, el número de dientes del cibario es la forma mas común de anomalía. El espécimen aqui descrito constituye un raro registro de anomalía.

Acute toxicity assessment of camphor in biopesticides by using Daphnia magna and Danio rerio

OBJECTIVES: An ecofriendly alternative to chemical pesticides is bio-pesticides, which are derived from natural sources. The interest in bio-pesticides is based on the disadvantages associated with chemical pesticides. METHODS: We conducted acute toxicity assessments of camphor, a major component of bio-pesticides, by using Daphnia magna (D. magna) as well as assessed the morphological abnormalities that occurred in Danio rerio (D. rerio) embryos. RESULTS: The median effective concentration of camphor on D. magna after 48 hours was 395.0 muM, and the median lethal concentration on D. rerio embryos after 96 hours was 838.6 muM. The no observed effect concentration and predicted no effect concentration of camphor on D. magna, which was more sensitive than D. rerio, were calculated as 55.2 muM and 3.95 muM, respectively. Morphological abnormalities in D. rerio embryos exposed to camphor increased over time. Coagulation, delayed hatching, yolk sac edema, pericardial edema, and pigmentation of embryos mainly appeared between 24 and 48 hours. Further, symptoms of scoliosis and head edema occurred after 72 hours. In addition, bent tails, ocular defects and collapsed symptoms of fertilized embryonic tissue were observed after 96 hours. CONCLUSIONS: The camphor toxicity results suggest that continuous observations on the ecosystem are necessary to monitor toxicity in areas where biological pesticides containing camphor are sprayed.

Morphological abnormalities and apoptosis in lamellar tissue of equines after intestinal obstruction and treatment with hydrocortisone / Alterações morfológicas e apoptose em tecido laminar de equinos após obstrução intestinal e tratamento com hidrocortisona

Four experimental groups of equines were used in order to study morphological abnormalities and apoptosis in lamellar tissue. Group Cg (control) was composed of animals without any surgical procedure; group Ig (instrumented), animals that underwent enterotomy; group Tg (treated), animals that were subjected to intestinal obstruction and were treated with hydrocortisone; and group Ug (untreated), animals that were subjected to intestinal obstruction without treatment. The lamellar tissue was analyzed regarding the presence of tissue abnormalities and apoptosis. No morphological abnormalities were observed in animals of surgical groups, and no difference in apoptosis was observed between groups. It was concluded that intestinal obstruction allowed laminitis to develop, probably by systemic activation, and that the maneuvers performed in the enterotomy aggravated the process. Hydrocortisone did not aggravate the lesions of the lamellar tissue.

Foram utilizados quatro grupos de equinos para estudar alterações morfológicas e apoptose no tecido lamelar. O grupo CG (controle) foi composto por animais sem o procedimento cirúrgico; o grupo Ig (instrumentado), por animais submetidos à enterotomia; o grupo Tg (tratado), por animais submetidos à obstrução intestinal e tratados com hidrocortisona; e o grupo Ug (não tratado), por animais submetidos à obstrução intestinal, sem tratamento. O tecido laminar foi analisado quanto à presença de alterações morfológicas e de apoptose. Foram observadas alterações morfológicas nos equinos submetidos a procedimento cirúrgico, mas não houve diferença entre os grupos com relação às células apoptóticas. Conclui-se que a obstrução intestinal permite o desenvolvimento da laminite provavelmente por ativação sistêmica e que as manobras realizadas na enterotomia podem ser consideradas como agravantes no processo. A hidrocortisona não agravou as lesões do tecido laminar.

Potential of biologically active plant oils to control mosquito larvae (Culex pipiens, Diptera: Culicidae) from an egyptian locality

The insecticidal effect of six commercially available plant oils was tested against 4th larval instars of Culex pipiens. Larvae were originally collected from Meit El-Attar, Qalyubia Governorate, Egypt, and then reared in the laboratory until F1 generation. The LC50 values were 32.42, 47.17, 71.37, 83.36, 86.06, and 152.94 ppm for fenugreek (Trigonella foenum-grecum), earth almond (Cyperus esculentus), mustard (Brassica compestris), olibanum (Boswellia serrata), rocket (Eruca sativa), and parsley (Carum ptroselinum), respectively. The tested oils altered some biological aspects of C. pipiens, for instance, developmental periods, pupation rates, and adult emergences. The lowest concentrations of olibanum and fenugreek oils caused remarkable prolongation of larval and pupal durations. Data also showed that the increase of concentrations was directly proportional to reduction in pupation rates and adult emergences. Remarkable decrease in pupation rate was achieved by mustard oil at 1000 ppm. Adult emergence was suppressed by earth almond and fenugreek oils at 25 ppm. In addition, the tested plant oils exhibited various morphological abnormalities on larvae, pupae, and adult stages. Consequently, fenugreek was the most potent oil and the major cause of malformation of both larval and pupal stages. Potency of the applied plant oils provided an excellent potential for controlling C. pipiens.

O efeito inseticida de seis óleos de plantas comercialmente disponíveis foi testado contra larvas de 4ºinstar de Culex pipiens. Larvas foram coletadas originalmente de Meit El-Attar, Qalyubia Governorate, Egito e então cultivadas no laboratório até a geração F1. Os valores LC50 foram 32,42, 47,17, 71,37, 83,36, 86,06 e 152,94 ppm para o feno grego (Trigonella foenum-grecum), amêndoa da terra (Cyperus esculentus), mostarda (Brassica compestris), olíbano (Boswellia serrata), rocket (Eruca sativa) e salsa (Carum ptroselium), respectivamente. Os óleos testados alteraram alguns aspectos biológicos do C. pipiens, por exemplo os períodos de desenvolvimento, estados de crisálida, e emergências de adultos. As concentrações mais baixas de óleo de olíbano e feno grego causaram extraordinário prolongamento da duração larval e pupal. Dados também mostraram que o aumento das concentrações foi diretamente proporcional à redução no estado de crisálida e emergências dos adultos. Notável decréscimo no estado de crisálida foi conseguido com o óleo de mostarda a 1000 ppm. Emergência de adulto foi diminuída no óleo de amêndoa da terra e feno grego a 25 ppm. Além do mais, os óleos de plantas testados, exibiram várias anormalidades morfológicas nas larvas, pupas e estádios adultos. Consequentemente, o óleo de feno grego foi o óleo mais potente e o maior causador de malformação em ambos estádios larval e pupal. Potencial dos óleos de plantas aplicados mostraram excelente resultado no controle do C. pipiens.