Myxobolus spp. (Cnidaria: Myxozoa) in introduced yellow perch Perca flavescens (Mitchill, 1814).

We surveyed introduced yellow perch Perca flavescens (Mitchill, 1814) from the Willamette River, OR, USA, to determine if these fish have co-introduced myxosporean parasites. Mature parasite myxospores were observed in brains of 3/19 fish, and were morphologically and molecularly consistent with Myxobolus neurophilus (Guilford 1963), a parasite known from yellow perch in their native range. We identified another Myxobolus species from the gill filaments of 1/22 fish. The spores from the gill filaments were oval-shaped, 11.7 (10.7-12.3) µm long × 8.6 (7.7-9.0) µm wide × 5.2 (4.6-5.6) µm thick, with two oval-shaped polar capsules 5.7 (5.1-6.5) µm × 2.7 (2.4-3.2) µm, each containing a polar tubule with 8-9 turns. Small-subunit ribosomal DNA sequences from each of four plasmodia were identical, and 4.0% different (over 1800 nucleotides) from the closest known myxosporeans. Interestingly, these sequences had overlapping peaks in their chromatograms, which suggested that DNA from multiple species was present. Hence, we isolated and sequenced three individual myxospores and found that they too had mixed chromatograms, which indicated presence of at least two sequence types of small-subunit ribosomal DNA in each spore (GenBank accession MK592012, MK592013), a rare character among described myxosporeans. The spore morphology, morphometry, tissue tropism, and DNA sequence supported a diagnosis of a novel species, Myxobolus doubleae n. sp. This parasite is unknown from yellow perch in its native range, despite extensive historical surveys, which suggests that introduced yellow perch might have acquired an endemic Myxobolus species via spillback from another fish host.

Host-parasite relationship of Ortholinea lauquen sp. nov. (Cnidaria: Myxozoa) and the fish Galaxias maculatus in northwestern Patagonia, Argentina.

Galaxias maculatus (Jenyns, 1842) is a widespread freshwater fish and an important component of the economically important whitebait fisheries across the Southern Hemisphere. We report a new myxosporean parasite (Cnidaria: Myxozoa) infecting the kidney of G. maculatus from northwestern Patagonia (Argentina). Ortholinea lauquen sp. nov. was characterized using myxospore morphology, morphometrics and small subunit rDNA (ssrDNA) sequence data. Our ssrDNA phylogenetic analyses showed that O. lauquen sp. nov. is a member of the oligochaete-freshwater urinary tract clade and basal to a clade containing 4 different spore morphotypes (Chloromyxum, Myxidium, Zschokkella, Hoferellus). We explored host-parasite relationships at the macro- and microscale by analyzing the distribution, tissue tropism and pathology of O. lauquen sp. nov. Prevalence was relatively low (7%) by microscopy, but PCR detection revealed hidden levels of infection (49%), with the highest detection in lakes Morenito and Moreno (63-90%, Río Negro Province). The only locality negative by both microscopy and PCR was the Caleufu River (Neuquén Province), suggesting differences in fish life history traits (landlocked vs. potamodromous) or preference of the putative obligate invertebrate host for lentic habitats. O. lauquen sp. nov. sporulates in the renal tubules and occasionally in the glomerular space. The plasmodia frequently occluded the tubule lumina, and cellular necrosis and disintegration of the epithelium were observed. O. lauquen sp. nov. could represent a potential threat to G. maculatus culture under intensive farming conditions.

CRISPR/Cas9 DNA cleavage at SNP-derived PAM enables both in vitro and in vivo KRT12 mutation-specific targeting.

CRISPR/Cas9-based therapeutics hold the possibility for permanent treatment of genetic disease. The potency and specificity of this system has been used to target dominantly inherited conditions caused by heterozygous missense mutations through inclusion of the mutated base in the short-guide RNA (sgRNA) sequence. This research evaluates a novel approach for targeting heterozygous single-nucleotide polymorphisms (SNPs) using CRISPR/Cas9. We determined that a mutation within KRT12, which causes Meesmann's epithelial corneal dystrophy (MECD), leads to the occurrence of a novel protospacer adjacent motif (PAM). We designed an sgRNA complementary to the sequence adjacent to this SNP-derived PAM and evaluated its potency and allele specificity both in vitro and in vivo. This sgRNA was found to be highly effective at reducing the expression of mutant KRT12 mRNA and protein in vitro. To assess its activity in vivo we injected a combined Cas9/sgRNA expression construct into the corneal stroma of a humanized MECD mouse model. Sequence analysis of corneal genomic DNA revealed non-homologous end-joining repair resulting in frame-shifting deletions within the mutant KRT12 allele. This study is the first to demonstrate in vivo gene editing of a heterozygous disease-causing SNP that results in a novel PAM, further highlighting the potential for CRISPR/Cas9-based therapeutics.

Erection of Ceratonova n. gen. (Myxosporea: Ceratomyxidae) to encompass freshwater species C. gasterostea n. sp. from threespine stickleback (Gasterosteus aculeatus) and C. shasta n. comb. from salmonid fishes.

Ceratonova gasterostea n. gen. n. sp. is described from the intestine of freshwater Gasterosteus aculeatus L. from the Klamath River, California. Myxospores are arcuate, 22.4 ± 2.6 µm thick, 5.2 ± 0.4 µm long, posterior angle 45° ± 24°, with 2 sub-spherical polar capsules, diameter 2.3 ± 0.2 µm, which lie adjacent to the suture. Its ribosomal small subunit sequence was most similar to an intestinal parasite of salmonid fishes, Ceratomyxa shasta (97%, 1,671/1,692 nucleotides), and distinct from all other Ceratomyxa species (<85%), which are typically coelozoic parasites in the gall bladder or urinary system of marine fishes. We propose erection of genus Ceratonova to contain both intestinal, freshwater species and reassign the salmonid parasite as Ceratonova shasta n. comb.

Complete life cycle of Myxobolus rotundus (Myxosporea: Myxobolidae), a gill myxozoan of common bream Abramis brama.

The life cycle of Myxobolus rotundus Nemeczek, 1911, a myxosporean parasite of the gills of common bream Abramis brama L., was studied under laboratory conditions. Mature Myxobolus spp. spores from plasmodia in the gills of wild bream were used to infect naïve oligochaete worms in a flow-through system of aquaria. Triactinomyxon-type actinospores were released from the oligochaetes 1 yr later and allowed to continually flow into a tank containing uninfected bream fry. The gills of the fry were checked for development of plasmodia in squash preparations 3 d postexposure, and then at weekly intervals for 8 wk. Tissue samples were fixed at each time point. Developing plasmodia were first observed 17 d post-exposure (Day 17). Mature spores were collected from plasmodia on Day 56 and were added to plastic dishes containing parasite-free Tubifex tubifex oligochaetes. Second-generation actinospores were released from these worms 8 mo post-exposure, and were morphologically identical to first-generation spores. Myxospores obtained from the bream fry were morphologically identical to those identified in wild bream as M. rotundus. Small subunit ribosomal RNA gene sequences obtained from first- and second-generation actinospores and the bream fry myxospores were 100% similar to M. rotundus spores from the original wild fish.

Renal myxozoanosis in weedy sea dragons, Phyllopteryx taeniolatus (Lacepède), caused by Sinuolinea phyllopteryxa n. sp.

Renal myxozoanosis was diagnosed histologically in 11 captive, wild caught, adult weedy (common) sea dragons, Phyllopteryx taeniolatus, from three separate public aquaria in the United States. Myxozoan spores were visible in wet mounts of kidney tissue and were associated with renal tubular dilatation and tubular epithelial cell hypertrophy. Light and electron microscopy revealed spore morphology consistent with the genus Sinuolinea. Spores were spheroidal, slightly dorso-ventrally compressed, length (L) 17.1 x width (W) 16.4 x thickness (T) 15.6 microm, with two shell valves joined at a distinct, sinuous sutural ridge, and had two nearly spherical polar capsules, L 5.5 x W 5.0 microm, with five to seven turns of the polar filament. There were no extra-valvular ridges or protrusions. DNA sequencing required the design of three new primers that yielded 1740 bp of 18S ribosomal DNA sequence. The parasite was determined to be novel based on morphological and molecular data, and was given the name Sinuolinea phyllopteryxa after its vertebrate host.

Expanded geographical distribution of Myxobolus cerebralis: first detections from Alaska.

The parasite responsible for salmonid whirling disease, Myxobolus cerebralis, was introduced to the USA in 1958. It has since spread across the country causing severe declines in wild trout populations, but has never been documented from Alaska. However, while assessing the risk of introduction of M. cerebralis into the state, we detected the parasite using a species-specific polymerase chain reaction (PCR) assay. Testing of 180 hatchery rainbow trout, Oncorhynchus mykiss (Walbaum), by pepsin trypsin digest (PTD) and quantitative PCR (QPCR) revealed 14 positive samples. Infection was confirmed by sequencing the parasite 18S rRNA gene and by a nested PCR assay based on the same gene. Sequence comparison of M. cerebralis from several locations demonstrated the Alaska isolates were genetically distinct and therefore not false-positives arising from contamination during processing. We were unable to visually identify myxospores, indicating that either infection was light or mature spores had not formed. A reference set of fish samples spiked with known numbers of myxospores verified the QPCR and PTD results. This paper presents DNA sequence data from the Alaska M. cerebralis isolates, provides a brief history of the fish and facility of origin, and discusses implications of different testing methods on asymptomatic fish populations.

The life cycle of Chloromyxum auratum (Myxozoa) from goldfish, Carassius auratus (L.), involves an antonactinomyxon actinospore.

The myxozoan parasite Chloromyxum auratumHallett, Atkinson, Holt, Banner & Bartholomew, 2006, was shown experimentally to have a two-host life cycle which involved a previously undescribed antonactinomyxon actinospore stage. Myxospores obtained from gall bladders of naturally infected feral goldfish, Carassius auratus (L.), were used to infect samples of mixed species of oligochaete worms obtained from the same locality as the fish: Fern Ridge Dam, Oregon, USA. After some 110 days post-exposure, actinospores were detected from the water above the oligochaetes. The 18S rDNA sequence of these actinospores was identical to the original myxospores. Spore release was sporadic, of low intensity and short duration, which confounded efforts to identify the host oligochaete species and infect naïve fish. This is the first life cycle that incorporates an actinospore of the collective group Antonactinomyxon, and the first life cycle demonstrated in the laboratory for a species of Chloromyxum.

Can patients sexually harass their physicians?

It is the fate of certain fashionable legal terms that capture the attention of the media to have their usage expanded beyond the contexts for which they were originally designed. Such is the case with the term sexual harassment. Essentially, it describes situations in which a powerful person attempts to influence an individual's economic or academic status based on his or her response to sexual comments or behaviors. Title VII and Title IX of the US Code contain federal laws that prohibit discrimination based on sex in the workplace and in the education system, respectively. Accordingly, sexual harassment that occurs within the context of the employment or academic arena is prohibited under Title VII and Title IX and has evolved to apply to hostile work or academic environments that do not per se influence an individual's economic or academic status.

Grieving and loss in parents with a schizophrenic child.

OBJECTIVE: The purpose of this study was to compare grief among parents who had an adult child with schizophrenia and parents who had "lost" an adult child through death or a head injury that resulted in an organic personality disorder. METHOD: Twenty-five parents from each group were studied. Self-report scales that assessed grief, depression, anxiety, and substance abuse were administered. RESULTS: There were significant differences with regard to grieving reactions and substance abuse. The parents with schizophrenic children had more ongoing grieving, and the parents of children with head injuries had more substance abuse. CONCLUSIONS: Parental loss of a child through schizophrenia leads to a pattern of chronic grief.

Electroconvulsive therapy revisited.

Electroconvulsive therapy (ECT) remains an extremely valuable tool in the treatment of certain psychiatric disorders. Despite the sensationalism of the past decade, ECT is the treatment of choice in severe depression that is unresponsive to other therapies or in patients who are overtly suicidal. In addition, ECT is particularly valuable in the elderly population, who are at risk for serious complications from the side effects associated with drug therapy. This report describes a case of recurrent depression in which ECT was successfully used.

Hypoalbuminemia may predispose infants to necrotizing enterocolitis.

Numerous risk factors for necrotizing enterocolitis (NEC) including prematurity, bowel ischemia, pathogenic bacteria, and hyperosmolar feedings have been proposed. Recent studies have demonstrated feeding intolerance and bowel dysfunction in children with hypoalbuminemia. No association between hypoalbuminemia and NEC has been suggested. The records of 45 patients with NEC and complete documentation of prenatal and birth histories were reviewed. A control (CONT) group of 90 children matched for maternal age (+/- 1 year), parity, gestational age (+/- 1 week), birth weight (+/- 20 g), type of delivery, sex, race, type of initial feeding, and perinatal stress was compiled. While all other measured parameters were similar in the two groups, premorbid albumin was significantly lower in the patients who subsequently developed NEC (P less than .001). These data suggest that newborns with hypoalbuminemia may have an increased risk of developing NEC.

Clinical characteristics, pathophysiology, and management of hydrocarbon ingestion: case report and review of the literature.

Accidental ingestion of hydrocarbons is an important cause of childhood poisoning. Due to the number of hydrocarbon products available as solvents, fuels, and cleaning agents, increased awareness is necessary on the part of health caretakers. The scope of complications involving the respiratory system in petroleum products ingestion is frequently overlooked. Physicians may thus apply standard therapeutic modalities used in treating common poisonings to the child who drank a petroleum distillate. Prompt recognition of presenting symptoms and understanding of pathophysiology are important to planning and providing treatment. The two cases of hydrocarbon ingestion reported in this paper illustrate the wide spectrum of problems associated with this condition. The pathophysiology, current management, and a review of the literature of hydrocarbon ingestion are presented.