Improved stool concentration procedure for detection of Cryptosporidium oocysts in fecal specimens.

Epidemiologic and laboratory data suggest that coprodiagnostic methods may fail to detect Cryptosporidium oocysts in stool specimens of infected patients. To improve the efficacy of stool concentration procedures, we modified different steps of the Formalin-ethyl acetate (FEA) stool concentration technique and evaluated these modifications by examining stool samples seeded with known numbers of Cryptosporidium oocysts. Because these modifications failed to improve oocyst detection, we developed a new stool concentration technique that includes FEA sedimentation followed by layering and flotation over hypertonic sodium chloride solution to separate parasites from stool debris. Compared with the standard FEA procedure, this technique improved Cryptosporidium oocyst detection. The sensitivities of the two concentration techniques were similar for diarrheal (watery) stool specimens (100% of watery stool specimens seeded with 5,000 oocysts per g of stool were identified as positive by the new technique, compared with 90% of stools processed by the standard FEA technique). However, the most significant improvement in diagnosis occurred with formed stool specimens that were not fatty; 70 to 90% of formed stool specimens seeded with 5,000 oocysts were identified as positive by the new technique, compared with 0% of specimens processed by the standard FEA technique. One hundred percent of formed specimens seeded with 10,000 oocysts were correctly diagnosed by using the new technique, while 0 to 60% of specimens processed by the standard FEA technique were found positive. Similarly, only 50 to 90% of stool specimens seeded with 50,000 oocysts were identified as positive by using the standard FEA technique, compared with a 100% positive rate by the new technique. The new stool concentration procedure provides enhanced detection of Cryptosporidium oocysts in all stool samples.     

Giardia transmission in a swimming pool.

In the fall of 1985, an outbreak of giardiasis occurred among several swimming groups at an indoor pool in northeast New Jersey. Nine clinical cases were identified, eight of whom had Giardia positive stool specimens. All were female; seven were adults (greater than 18 years) and two were children. The attack rate was highest (39 per cent, 5/13) for the ladies lap group who had exposure on one day. These cases had no direct contact with children or other risk factors for acquiring Giardia. Infection most likely occurred following the ingestion of swimming pool water contaminated with Giardia cysts. The source of Giardia contamination was a handicapped child who had a fecal accident in the pool. He was a member of a group that swam at the same time as the ladies lap group. A stool survey of the handicapped group showed that of the 20 persons tested, nine were positive for Giardia, including the specimen from this child. Examination of the pool records showed that no chlorine levels had been taken on the day of the fecal accident and that on the following day the chlorine level was zero. This is the second report of Giardia transmission among swimming pool attendees. It emphasizes the need to maintain appropriate chlorine levels in swimming pools and to institute measures to clear pools after a fecal accident.     

Recommendations for collection of laboratory specimens associated with outbreaks of gastroenteritis

Recent discoveries have implicated a number of "new" (i.e., previously unrecognized) infectious agents as important causes of outbreaks of gastroenteritis. Unfortunately, the ability to detect these agents in an outbreak can be limited by two factors: 1) the lack of appropriate assays-many of which are still in developmental stages and are not readily available to clinical laboratories, and 2) inadequately or improperly collected specimens. At CDC, many newly developed assays are being used for research and for outbreak investigations. The information in this report is especially intended for public health agencies that collaborate with CDC in investigating outbreaks of gastroenteritis. The report provides an update on guidelines and recommendations for the proper collection of specimens to be sent to CDC, gives general background information concerning some recently discovered pathogens, lists some of the tests available at CDC, and provides a list of CDC contacts. The guidelines and the general information provided on causes of outbreaks of gastroenteritis can be also used by public health workers for investigations when specific testing is available and appropriate.     

Efficacy of 1% permethrin for the treatment of head louse infestations among Kosovar refugees

We assessed the prevalence of head louse infestation and the effectiveness of 1% permethrin against head lice in Kosovar refugees. A currently infested case was defined as a person with observable crawling lice (adults or nymphs) or a person with nits on the hair shaft within a quarter-inch of the scalp. Of the 1,051 refugees screened upon arrival in the United States, 107 (10%) were infested. Crawling lice (adults or nymphs) were observed on 62 (6%) of the individuals examined. Refugees with crawling lice were treated with a pediculicide containing 1% permethrin. Of these, 57 were reexamined the next day. Twenty of the 57 individuals were reexamined 7 days after treatment. No crawling lice were found on any of the refugees examined after treatment. We conclude that 1% permethrin treatment was effective in louse control in this refugee population.     

Autochthonous dermal leishmaniasis in Texas.

Over a 2-year period two cases of dermal leishmaniasis of special interest were recognized in Texas. The first occurred in 1972 in a 74-year-old woman who had residences in Dilworth and Gahzales, Gonzales County, and the other was in a 56-year-old man from Kenedy, Karnes County, in 1974. Both cases were biopsy- and culture-positive, and the second patient exhibited anti-Leishmania antibodies by indirect immunofluorescent antibody and direct agglutination tests. Epidemiologic investigation revealed no association between the two cases and suggested that both patients had acquired their infections locally in southern Texas. Moreover, serologic evidence of Leishmania infection was uncovered in a neighbor of the second case and in 3 dogs living nearby. Potential sylvatic reservoirs and arthropod vectors of the disease are resident in the area. Epidemiologic data suggest that dermal leishmaniasis is endemic in south-central Texas.     

Volumetric Analysis of Regional Variability in the Cerebellum of Children with Dyslexia

Cerebellar deficits and subsequent impairment in procedural learning may contribute to both motor difficulties and reading impairment in dyslexia. We used quantitative magnetic resonance imaging to investigate the role of regional variation in cerebellar anatomy in children with single-word decoding impairments (N?=?23), children with impairment in fluency alone (N?=?8), and typically developing children (N?=?16). Children with decoding impairments (dyslexia) demonstrated no statistically significant differences in overall grey and white matter volumes or cerebellar asymmetry; however, reduced volume in the anterior lobe of the cerebellum relative to typically developing children was observed. These results implicate cerebellar involvement in dyslexia and establish an important foundation for future research on the connectivity of the cerebellum and cortical regions typically associated with reading impairment.     

RAGE Deficiency Improves Postinjury Sciatic Nerve Regeneration in Type 1 Diabetic Mice

Peripheral neuropathy and insensate limbs and digits cause significant morbidity in diabetic individuals. Previous studies showed that deletion of the receptor for advanced end-glycation products (RAGE) in mice was protective in long-term diabetic neuropathy. Here, we tested the hypothesis that RAGE suppresses effective axonal regeneration in superimposed acute peripheral nerve injury attributable to tissue-damaging inflammatory responses. We report that deletion of RAGE, particularly in diabetic mice, resulted in significantly higher myelinated fiber densities and conduction velocities consequent to acute sciatic nerve crush compared with wild-type control animals. Consistent with key roles for RAGE-dependent inflammation, reconstitution of diabetic wild-type mice with RAGE-null versus wild-type bone marrow resulted in significantly improved axonal regeneration and restoration of function. Diabetic RAGE-null mice displayed higher numbers of invading macrophages in the nerve segments postcrush compared with wild-type animals, and these macrophages in diabetic RAGE-null mice displayed greater M2 polarization. In vitro, treatment of wild-type bone marrow–derived macrophages with advanced glycation end products (AGEs), which accumulate in diabetic nerve tissue, increased M1 and decreased M2 gene expression in a RAGE-dependent manner. Blockade of RAGE may be beneficial in the acute complications of diabetic neuropathy, at least in part, via upregulation of regeneration signals.Diabetes leads to the development of multiple complications (1–3). Peripheral neuropathy affects 30–50% of all diabetic patients (4–6). Individuals with diabetes are more vulnerable to superimposed thermal and pressure injuries (7–10). Diabetic individuals exposed to either topical application of capsaicin or intracutaneous excision axotomy (punch skin biopsy) displayed a reduction in regenerative rate, even without evidence of neuropathy, and reduced axonal regenerative sprouting and blood vessel growth, respectively, compared with nondiabetic control subjects (11,12). Studies of diabetic animals reported a delay of axonal regeneration after acute sciatic nerve crush compared with nondiabetic mice (13). Evidence suggests that enhanced accumulation of advanced glycation end products (AGEs) may be an important contributing mechanism to the pathogenesis of diabetes complications (14,15). AGEs are a heterogeneous group of molecules that impact cellular properties and gene expression via specific receptors such as receptor for advanced end-glycation product (RAGE) (16–18). RAGE, a pattern recognition receptor, also interacts with multiple members of the proinflammatory S100/calgranulin family and with high-mobility group box 1 protein (HMGB1); both classes of molecules are implicated in inflammation and cellular migration (19,20). These non-AGE ligands may be released by dying cells, and evidence suggests that although RAGE is not intimately involved in innate immune responses, its upregulation and activation by these ligands contribute to sustained inflammation and suppression of repair (21,22).These considerations prompted us to hypothesize that RAGE action in superimposed acute injury to the peripheral nerve, particularly in diabetes, attenuates neurite outgrowth and axonal regeneration via tissue-damaging inflammatory mechanisms. We subjected wild-type (WT) and homozygous RAGE-null mice to acute sciatic nerve crush to dissect the specific contribution of bone marrow RAGE expression. We also subjected WT mice to lethal irradiation and performed reconstitution with bone marrow expressing or devoid of RAGE.     

A multiyear prospective study of the risk factors for and incidence of diarrheal illness in a cohort of Peace Corps volunteers in Guatemala

BACKGROUND: Diarrheal illness is the most common medical disorder among travelers from developed to developing countries and is common among expatriate residents in developing countries. OBJECTIVE: To assess the risk factors for and incidence of diarrheal illness among Americans living in a developing country. DESIGN: Prospective longitudinal study. SETTING: Rural Guatemala. PATIENTS: Cohort of 36 Peace Corps volunteers. MEASUREMENTS: Collection of daily dietary and symptom data for more than 2 years; identification by multivariate Poisson regression analyses of risk factors for clinically defined episodes of diarrheal illness. RESULTS: The 36 Peace Corps volunteers in this study had 307 diarrheal episodes (median, 7 per person), which lasted a median of 4 days (range, 1 to 112) and a total of 10.1% of the 23 689 person-days in the study. The incidence density (episodes per person-year) was 4.7 for the study as a whole, 6.1 for the first 6-month period, 5.2 for the second 6-month period, and 3.6 thereafter. Statistically significant risk factors for diarrheal illness included drinking water whose source (for example, the tap) and, therefore, quality, was unknown to the person; eating food prepared by a Guatemalan friend or family; eating food at a small, working-class restaurant; eating fruit peeled by someone other than a Peace Corps volunteer; drinking an iced beverage; and eating ice cream, ice milk, or flavored ices. The relative risks comparing the presence of these exposures during the first 6-month period overseas with their absence during the second year of residence ranged from 1.90 to 2.67, and the summary attributable risk percentage (that is, the percentage of diarrheal episodes that could be ascribed to the exposures) was 75.4%. Exposures generally were riskier if they occurred during travel elsewhere in Guatemala rather than in the person's usual work area. CONCLUSIONS: Diarrheal illness of mild-to-moderate severity continued to occur throughout Peace Corps service but decreased in incidence as length of stay increased. Various dietary behaviors increased the risk for diarrheal illness, which suggests that avoidance of potentially risky foods and beverages is beneficial.     

Relative Carnitine Deficiency in Autism

A random retrospective chart review was conducted to document serum carnitine levels on 100 children with autism. Concurrently drawn serum pyruvate, lactate, ammonia, and alanine levels were also available in many of these children. Values of free and total carnitine (p < 0.001), and pyruvate (p=0.006) were significantly reduced while ammonia and alanine levels were considerably elevated (p < 0.001) in our autistic subjects. The relative carnitine deficiency in these patients, accompanied by slight elevations in lactate and significant elevations in alanine and ammonia levels, is suggestive of mild mitochondrial dysfunction. It is hypothesized that a mitochondrial defect may be the origin of the carnitine deficiency in these autistic children.