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An essential and highly selective protein import pathway encoded by nucleus-forming phage.
Targeting proteins to specific subcellular destinations is essential in prokaryotes, eukaryotes, and the viruses that infect them. Chimalliviridae phages encapsulate their genomes in a nucleus-like replication compartment composed of the protein chimallin (ChmA) that excludes ribosomes and decouples transcription from translation. These phages selectively partition proteins between the phage nucleus and the bacterial cytoplasm. Currently, the genes and signals that govern selective protein import into the phage nucleus are unknown. Here, we identify two components of this protein import pathway: a species-specific surface-exposed region of a phage intranuclear protein required for nuclear entry and a conserved protein, PicA (Protein importer of chimalliviruses A), that facilitates cargo protein trafficking across the phage nuclear shell. We also identify a defective cargo protein that is targeted to PicA on the nuclear periphery but fails to enter the nucleus, providing insight into the mechanism of nuclear protein trafficking. Using CRISPRi-ART protein expression knockdown of PicA, we show that PicA is essential early in the chimallivirus replication cycle. Together, our results allow us to propose a multistep model for the Protein Import Chimallivirus pathway, where proteins are targeted to PicA by amino acids on their surface and then licensed by PicA for nuclear entry. The divergence in the selectivity of this pathway between closely related chimalliviruses implicates its role as a key player in the evolutionary arms race between competing phages and their hosts.
An essential and highly selective protein import pathway encoded by nucleus-forming phage.
UNLABELLED: Targeting proteins to specific subcellular destinations is essential in prokaryotes, eukaryotes, and the viruses that infect them. Chimalliviridae phages encapsulate their genomes in a nucleus-like replication compartment composed of the protein chimallin (ChmA) that excludes ribosomes and decouples transcription from translation. These phages selectively partition proteins between the phage nucleus and the bacterial cytoplasm. Currently, the genes and signals that govern selective protein import into the phage nucleus are unknown. Here we identify two components of this novel protein import pathway: a species-specific surface-exposed region of a phage intranuclear protein required for nuclear entry and a conserved protein, PicA, that facilitates cargo protein trafficking across the phage nuclear shell. We also identify a defective cargo protein that is targeted to PicA on the nuclear periphery but fails to enter the nucleus, providing insight into the mechanism of nuclear protein trafficking. Using CRISPRi-ART protein expression knockdown of PicA, we show that PicA is essential early in the chimallivirus replication cycle. Together our results allow us to propose a multistep model for the Protein Import Chimallivirus (PIC) pathway, where proteins are targeted to PicA by amino acids on their surface, and then licensed by PicA for nuclear entry. The divergence in the selectivity of this pathway between closely-related chimalliviruses implicates its role as a key player in the evolutionary arms race between competing phages and their hosts. SIGNIFICANCE STATEMENT: The phage nucleus is an enclosed replication compartment built by Chimalliviridae phages that, similar to the eukaryotic nucleus, separates transcription from translation and selectively imports certain proteins. This allows the phage to concentrate proteins required for DNA replication and transcription while excluding DNA-targeting host defense proteins. However, the mechanism of selective trafficking into the phage nucleus is currently unknown. Here we determine the region of a phage nuclear protein that targets it for nuclear import and identify a conserved, essential nuclear shell-associated protein that plays a key role in this process. This work provides the first mechanistic model of selective import into the phage nucleus.
[Necrotizing enteritis and geophagia].
Two cases of intestinal perforation due to necrotising enteritis in patients with pica are presented. The aetiopathogenesis of pica is discussed as well as its role in the development of necrotising enteritis.
Some observations on aneurysms of the proximal internal carotid artery.
The author reports on 41 aneurysms of the proximal internal carotid artery (PICA) demonstrated in 36 patients with subarachnoid hemorrhage. The patients included a striking preponderance of women, and there was a high incidence of multiple aneurysms. In cases with multiple aneurysms the PICA aneurysm was usually found incidentally, a more distal aneurysm on the internal carotid artery being the source of hemorrhage. An infundibulum at the origin of a posterior communicating artery was unusually common in these patients. The origin of the ophthalmic artery is proposed as the angiographic landmark of the level at which the internal carotid artery penetrates the dura mater.
Visceral larva migrans and eosinophilia in an emotionally disturbed child.
Visceral larva migrans, Entamoeba coli, evidence of latent toxoplasmosis and a history of plumbism were found in an emotionally disturbed, retarded child. Patients with pica should be screened for parasitism and other diseases transmitted orally.
Geophagic lead nephropathy: case report.
An adult presenting with anemia and seizures was found to have lead poisoning. Chelation therapy undertaken before the source of exposure was known was accompanied by clinical improvement. Recurrence of an excessive body lead burden despite chelation led to the discovery of pica for lead-contaminated garden soil. Lead nephropathy progressed when the geophagia was resumed.
Epidemiological characteristics and clinical findings in patients with serologically proven toxocariasis.
The enzyme-linked immunosorbent assay (ELISA) using Toxocara canis larva-specific antigens was used to confirm a presumptive clinical diagnosis of visceral larva migrans in patients whose serum specimens were submitted to the Center for Disease Control. The clinical and epidemiological characteristics of patients with diagnostic ELISA titres (greater than or equal to log2 5) were compared with patients who had less or no detectable antibody. The typical patient with serologically confirmed toxocaral visceral larva migrans was about five years of age and resided in the southern half of the USA. Clinical findings were likely to include leucocytosis, eosinophilia, an increased anti-A or anti-B isohaemagglutinin titre, and an elevated serum IgG level. Pica, as ascertained by a physician questionnaire, was not a consistent finding and both sexes were nearly equally represented.
Extraction of lead from printed matter at physiological values of pH.
In an in vitro laboratory study of the extractability of lead in printed matter it was found that dangerous quantities of lead, up to 200 mug, could be extracted from "small" pieces of printed paper at pH values in the range of human gastric fluid. Lead was not extracted at pH values in the range of human saliva. Children who chew printed matter may not be in danger of absorbing lead, but the pica-prone child who swallows printed material may be at risk of absorbing excessive amounts of this toxic metal. The use of printing inks containing high-lead levels should be discouraged.
Lead toxicity and nutritional deficiencies.
Under appropriate conditions, deficiencies of certain minerals and vitamins as well as high intakes of dietary fat increase the toxicity of a given dose of lead in experimental animals. The severity of lead poisoning can also be increased by the consumption of either deficient or excessive levels of protein. Mineral deficiencies appear to have some of the most profound effects on lead toxicity, since the consequences of plumbism can be exaggerated by feeding diets low in calcium, phosphorus, iron, zinc, and in some cases, copper. Evidence for an antagonism between lead and nutritional levels of selenium is inconclusive. Vitamin E deficiency and lead poisoning interact to produce an anemia in rats that is more severe than that caused by either treatment alone. Lead apparently exerts a pro-oxidant stress on the red cell, thereby causing its accelerated destruction. One of the biochemical mechanisms of lead poisoning may be the disruption of normal membrane architecture, thereby leading to peroxidative damage. Epidemiological surveys have suggested a negative correlation between the poor nutritional status of children with regard to calcium and the concentration of lead in blood. Other examples of potential interactions of mineral status and lead poisoning in humans include the hypothesized hazards of soft water to public health in areas with lead plumbing and the possible role of mineral deficiencies in the etiology of pica. Experimental studies have shown that in some situations combined nutritional deficiencies can have an additive effect in potentiating lead toxicity.
Effect of a low chloride diet on lactating Holstein cows.
Twenty primiparous lactating Holstein cows were fed diets supplemented with either .5% sodium chloride or .75% sodium bicarbonate. Those fed the latter diet conserved chloride by reducing the chloride concentration in urine, feces, and, to a small but nonsignificant amount, in milk. The low chloride diet did not affect consumption of feed dry matter and water or milk production and composition. There was no indication of pica or other aberrant taste behavior. One half of the cows in each diet group was given free choice salt block of trace elements. Those fed the low chloride diet consumed more salt (337 versus 149 g/cow per wk). When chloride requirements have been established and data on chloride composition become more widely available, it will be possible to include both sodium and chloride in diet formulations and omit salt. This will reduce the amount of salt needed as either a sodium or chloride supplement.
Clinical, serologic, and epidemiologic characteristics of ocular toxocariasis.
The clinical, serologic, and epidemiologic characteristics of 17 cases of ocular toxocariasis (OT) were studied and compared with those of a control group of 15 cases of other ocular diseases whose differential diagnosis included retinoblastoma. The prevalence and mean titers of Toxocara antibody detected by enzyme-linked immunosorbent assay were greater (P less than 0.005) for patients with OT than for the control group, but not all clinically diagnosed OT cases had detectable antibody. The prevalence of pica was significantly greater in cases than in controls (P less than 0.05). Almost all case and control patients had a history of exposure to pet dogs and cats, but recent exposure to puppies (less than 3 months old) was significantly associated with Toxocara infection in this study group.
Visceral larva migrans. A review and reassessment indicating two forms of clinical expression: visceral and ocular.
Visceral larva migrans is a disease in which the larvae of animal parasites invade human tissues but do not complete their life cycles. The most frequent cause of this illness in children is the dog roundworm, Toxocara canis. A review of the literature, as well as our clinical experience, indicates that there are two forms of clinical expression: one, visceral, and the other, ocular. In general the clinical and laboratory abnormalities (hepatomegaly, recurrent pneumonia, eosinophilia, and hyperglobulinemia) usually associated with visceral disease are absent in children with ocular abnormalities. Conversely, there is a general lack of eye complications in patients with systemic disease. Reasons for these variations in clinical expression are unknown, but immune responses of the host and the antigenic composition of the parasite may be contributing factors.
Morbidity pattern among children below 5 years in an urban Sindhi community.
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alpha-Amylase activity of human neonate and adult saliva.
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Geophagia in response to stress and arthritis.
Geophagia, the consumption of nonnutritive dirt, has previously been shown to be increased when rats have been made acutely ill. The present paper reports two experiments in which geophagia in response to stress and arthritis was measured. In the first experiment, experimental rats were injected with Freund's Complete Adjuvant (FCA), a mineral oil suspension of killed Mycobacterium butyricum. They ate significantly more kaolin (a nonnutritive substance) than controls, suggesting that geophagia is a behavioral measure of illness. In a second study, kaolin consumption was similarly elevated in stressed and/or arthritic animals as well as in control rats living in the same room. Nonstressed controls housed in a remote room ate significantly less kaolin than rats in all other groups. It was concluded that geophagia may occur in response to any homeostatic alteration (stress state). Furthermore, this state can presumably be communicated to naive rats from adjacently-housed stressed animals. Geophagia was postulated to occur in animals who experience a generalized stress reponse since this state may involve gastro-intestinal malaise.
Behavioral differences between two populations of wild rats: implications for domestication research.
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Neuropsychological dysfunction in children with chronic low-level lead absorption.
To investigate the relation between low-level absorption and neuropsychological function, blind evaluations were under-taken in forty-six symptom-free children aged 3-15 years with blood-lead concentrations of 40-68 mug. per 100 ml. (mean 48 mug. per 100 ml.) and in seventy-eight ethnically and socioeconomically similar controls with levels greater than mug. per 100 ml. (mean 27 mug. per (100 ml). All children lived within 6-6 km. of a large, lead-emitting smelter, and in many cases residence there had been lifelong. Mean age in the lead group was 8-3 years and in the controls 9-3. Testing with Wechsler intelligence scales for schoolchildren and preschool children (W.I.S.C. and W.P.P.S.I.) showed age-adjusted performance I.Q. to be significantly decreased in the group with higher lead levels (mean scores, W.I.S.C. plus W.P.P.S.I., 95 v. 103). Children in all ages in the lead group also had significant slowing in a finger-wrist tapping test. Full-scale I.Q., verbal I.Q., BEHAVIOUR, AND HYPERACTIVITY RATINGS DID NOT DIFFER.