The role of peripheral blood smear examination in the evaluation of suspected platelet-related disorders in children: A practical approach and an illustrated review.

Platelets, along with coagulation factors and vasculature, represent the three main compartments of hemostasis. Upon investigation of a suspected hemostasis disorder, platelet count, size and morphology often offer important clues to the diagnosis or help narrow the differential diagnosis. In this review, we describe a general approach to diagnosing platelet disorders, starting with easily obtained data such as findings of complete blood count (CBC) and microscopic review of a stained peripheral blood smear. We discuss general findings that help separate consumptive from underproduction thrombocytopenia. We further touch on inherited thrombocytopenia disorders after classifying them into those associated with small, normal sized or large platelets. Illustrative microscopic images are provided where contributory. We conclude with a suggested algorithmic step-by-step approach to investigating a suspected platelet disorder in children.

Comparison of aflatoxigenic and nonaflatoxigenic isolates of Aspergillus flavus using DNA amplification fingerprinting techniques.

Aspergillus flavus is a filamentous fungus that produces mycotoxins in many food and feed crops, such as maize (Zea mays L.). Isolates were analyzed for toxin production by nucleic acid profiles in an attempt to differentiate aflatoxigenic from nonaflatoxigenic isolates. A total of 41 aflatoxigenic and 34 nonalfatoxigenic isolates were included in the study. The isolates were evaluated initially using DNA amplification fingerprinting (DAF) without clear resolution of the groups. A weak association of aflatoxigenic isolates was observed, as evidenced by their clustering in 18 of 81 trees recovered from maximum parsimony analysis of binary characters derived from arbitrary signatures from amplification profiles (ASAP) data; nonaflatoxigenic isolates exhibited a pattern of paraphyletic laddering. Up to five markers unambiguously supported the aflatoxigenic isolate grouping, but the presence of alternative conflicting topologies in equally parsimonious trees precluded the observation of meaningful statistical support. With additional markers for genome of A. flavus, this method could be used to resolve toxigenic from nontoxigenic strains. This additional work could resolve aflatoxigenic isolates of A. flavus present on maize plants using ASAP, which would reduce labor intense costs and potentially lead to faster determination of resistant cultivars in breeding efforts.

Effects of nematicides on cotton root mycobiota.

Baseline information on the diversity and population densities of fungi collected from soil debris and cotton (Gossypium hirsutum L.) roots was determined. Samples were collected from Tifton, GA, and Starkville, MS containing cotton field soil treated with the nematicides 1,3-dichloroproprene (fumigant) and aldicarb (granules). A total of 10,550 and 13,450 fungal isolates were collected from these two study sites, respectively. Of this total, 34 genera of plant pathogenic or saprophytic species were identified. Pathogenic root fungi included Fusarium spp. (40% of all isolations), Macrophomina, Pythium, Rhizoctonia, and Sclerotium. Fusarium and Rhizoctonia were the most common fungal species identified and included F. oxysporum, F. verticillioides and F. solani, the three Fusarium species pathogenic on cotton plants. Population densities of Fusarium were not significantly different among locations or tissue types sampled. Macrophomina was isolated at greater numbers near the end of the growing seasons. Anastomosis groups of R. solani isolated from roots and soil debris included AG-3, -4, -7, 2-2, and -13 and anastomosis groups of binucleate Rhizoctonia included CAG-2, -3, and -5. Occurrences and frequency of isolations among sampling dates were not consistent. Fluctuations in the frequency of isolation of Rhizoctonia did not correspond with changes in frequency of isolation of the biological control fungus, Trichoderma. When individual or pooled frequencies of the mycobiota were compared to nematicide treatments, no specific trends occurred between treatments, application methods or rates. Results from this study show that use of 1,3-D and aldicarb in cotton fields does not significantly impact plant pathogenic fungi or saprophytic fungal populations. Thus cotton producers need not adjust seedling disease control measures when these two nematicides are used.

Pod and seed mycoflora on transgenic and conventional soybean [Glycine max (L.) Merrill] cultivars in Mississippi.

A 2-year (1999-2000) study was conducted at Starkville and Stoneville, MS to determine if the occurrence of the mycoflora varied on Roundup Ready (transgenic) compared to conventional soybean (Glycine max) cultivars. A total of 7,658 fungal isolates were identified from the pod and seed tissues of four cultivars compared at growth stages R6 and R8. Ninety-nine percent of all fungi isolated were mitosporic fungi and ascomycetes. In both years, total fungal isolates from the two locations were greater from the pod (65%) than from seed (33%) tissues. Isolation frequency from conventional cultivars was 54% compared to 46% for the transgenic cultivars. The most common fungi identified that are reported pathogens of soybean included Alternaria, Cercospora, Cladosporium, Diaporthe, Fusarium and Verticillium spp. When main effects and interactions were compared among the frequency data for the fungal genera, significant differences occurred, but consistent trends were not noted. Isolation frequencies of Diaporthe spp. during the R6 growth stage, were significantly greater on the conventional than on the transgenic cultivars in both years of the study, but only at Starkville. Isolation frequencies from samples taken during the R8 growth stage were similar at both locations in 1999 and 2000. Fusarium spp. isolated at R6 and R8 growth stages from pod and seed tissues were significantly greater on conventional than on transgenic cultivars in 2000. Even though frequencies were often significantly different between the transgenic and conventional cultivars, the data was not consistent between locations, pod and seed tissues, or growth stages. The pod and seed mycoflora of transgenic and conventional soybean cultivars was, therefore, similar in Mississippi.

First Report of Rhizoctonia solani AG-7 on Cotton in Mississippi.

During a field study of the soilborne mycobiota on cotton (Gossypium hirsutum L.) roots, Rhizoctonia solani Kühn AG-7 was isolated from dark brown lesions present on the tap, feeder, and secondary roots onto potato-dextrose agar (PDA) (34 g of medium per liter of distilled water). Isolate identification was confirmed using tester R. solani AG-7 isolates (Carling, University of Alaska) for comparison during anastomosis pairing. To confirm pathogenicity, six AG-7 isolates (65, 66, 67, 68, 69, 70) obtained from cotton roots were tested in a plant growth incubator (18-24°C) by mixing 2.5 ml of 2-week-old cornmeal sand inoculum (3 g cornmeal, 100 g sand, and 20 ml distilled water) with 500 ml of autoclaved soil into each of five replicate polystyrene pots (15 cm wide × 20 cm long) per isolate. Five control pots containing noninfested soil were added for comparison. Five seed of cv. Deltapine 50 were sown into each pot. For all six isolates, mean stand counts (ranging from 0 and 1.3 plants per pot) were significantly lower (P ≤ 0.05) compared with those of plants in noninfested pots (3.8 plants per pot) at 21 days after planting. Furthermore, when the roots of surviving seedlings were evaluated for disease severity, brown, discolored lesions were observed at the base of stems and on roots in infested pots for all six isolates. Six AG-7 cultures of the pathogen were reisolated from symptomatic tissues onto PDA. The experiment was repeated with similar results. This is the first report of AG-7 occurring in Mississippi.

Communicating intrathoracic hydrocele.

Tense ascites may cause herniation of parietal peritoneal reflection into the mediastinum at the gastroesophageal junction. This may produce a mass visible on chest radiograph and computed tomography (CT). This communicating intrathoracic hydrocele may occur in the absence of hiatal hernia and may be confused with other middle mediastinal fluid collections including necrotic tumor, abscess, cyst of foregut origin, or pancreatic pseudocyst. Recognition of this entity in patients with ascites should prevent diagnostic confusion and unnecessary further evaluation.

Hypercalcemia secondary to a pseudoparathormone-secreting ameloblastoma.

An unusual case of hypercalcemia associated with an ameloblastoma occurred. The sequence of recurring ameloblastoma, increased serum calcium levels and urinary cyclic adenosine monophosphate (cAMP) levels, removal of the tumor, and decrease of those levels to normal suggests that the ameloblastoma was secreting a parathormone-like substance that, in turn, was stimulating the production of nephrogenous cAMP and elevating the serum calcium level.