Vertically transmitted microbiome protects eggs from fungal infection and egg failure.

BACKGROUND: Beneficial microbes can be vertically transmitted from mother to offspring in many organisms. In oviparous animals, bacterial transfer to eggs may improve egg success by inhibiting fungal attachment and infection from pathogenic microbes in the nest environment. Vertical transfer of these egg-protective bacteria may be facilitated through behavioral mechanisms such as egg-tending, but many species do not provide parental care. Thus, an important mechanism of vertical transfer may be the passage of the egg through the maternal cloaca during oviposition itself. In this study, we examined how oviposition affects eggshell microbial communities, fungal attachment, hatch success, and offspring phenotype in the striped plateau lizard, Sceloporus virgatus, a species with no post-oviposition parental care. RESULTS: Relative to dissected eggs that did not pass through the cloaca, oviposited eggs had more bacteria and fewer fungal hyphae when examined with a scanning electron microscope. Using high throughput Illumina sequencing, we also found a difference in the bacterial communities of eggshells that did and did not pass through the cloaca, and the diversity of eggshell communities tended to correlate with maternal cloacal diversity only for oviposited eggs, and not for dissected eggs, indicating that vertical transmission of microbes is occurring. Further, we found that oviposited eggs had greater hatch success and led to larger offspring than those that were dissected. CONCLUSIONS: Overall, our results indicate that female S. virgatus lizards transfer beneficial microbes from their cloaca onto their eggs during oviposition, and that these microbes reduce fungal colonization and infection of eggs during incubation and increase female fitness. Cloacal transfer of egg-protective bacteria may be common among oviparous species, and may be especially advantageous to species that lack parental care.

Towards the understanding and prediction of material changes during micronisation using atomic force microscopy.

In this study we aim to explore the potential links between the mechanical properties, micronisation behaviour and surface energy of carbamazepine polymorphs using atomic force microscopy (AFM) measurements of material properties at the nanoscale. Carbamazepine Forms I, II and III were prepared and confirmed using X-ray powder diffraction (XRPD). AFM measurements of indentation hardness, Young's modulus and surface energy were made on the starting material. In addition, the surface energy was measured immediately after micronisation and after storage for four weeks. Carbamazepine polymorphs could be ranked by Young's modulus and hardness. Surface energy measurements showed an increase after micronisation in all cases, and a varying relaxation after storage for four weeks. Form I showed a smaller particle size distribution, indicating more complete micronisation. A promising correlation was observed between the hardness/Young's modulus ratio and the micronisation behaviour, in terms of particle size reduction and surface energy change. The results show potential for the predictive capacity of such an approach, and help to provide a greater understanding of material behaviour and properties during micronisation.

Management of thoracic empyema in childhood: does the pleural thickening matter?

AIMS: To determine the clinical course and long term outcome of empyema treated without decortication. METHODS: Fourteen consecutive admissions to one hospital were studied; radiological resolution and lung function were subsequently followed. The children were aged 2-14 years. All were treated with intravenous antibiotics and chest drain only. RESULTS: All patients had extensive pleural thickening evident on chest x ray examination at the time of discharge, which resolved entirely over a period of 2-16 months. Lung function was measured in 13 children, and showed no evidence of restrictive or obstructive deficit: mean (SD) values as per cent predicted for height were: forced expiratory volume in one second (FEV1) 107.5 (9.6), forced vital capacity (FVC) 95.5 (8.8), total lung capacity (TLC) 98.6 (20.7). Individual children all had values in the normal range (80-120 per cent predicted). CONCLUSION: Results suggest that decortication is not necessary in children to prevent long term problems with pleural thickening, and should be undertaken on the basis of the clinical picture (failure of fever resolution), rather than radiological appearance.

Optimizing pretransfusion antibody detection and identification: a parallel, blinded comparison of tube PEG, solid-phase, and automated methods.

BACKGROUND: The ideal pretransfusion testing strategy identifies maximal significant antibodies at minimal cost. Objectives of this study were to compare the characteristics of three testing methods and determine their optimal incorporation into the following generic testing sequence: 1) screen, for antibodies 2) if results are positive, use primary identification method, 3) if results are inconclusive, use secondary identification method. STUDY DESIGN AND METHODS: A total of 2000 consecutive, unselected, coded specimens were tested with three screening methods-PEG IAT, manual and automated solid phase red cell adherence assay (SPRCA). Of 202 positive results, 187 were of sufficient volume and were tested with both PEG and manual SPRCA identification panels, yielding 82 with significant antibodies, plus one that was negative by both methods found on retrospective review of nonstudy results. Calculations were made on the 1985 volume-sufficient specimens, simulating the possible testing permutations. RESULTS: Manual SPRCA was the most sensitive antibody screen (67/83 = 81%) and the least specific (1840/1902 = 97%); automated SPRCA was the least sensitive (54/83 = 65%) and most specific (1883/1902 = 99%); and PEG was intermediate in both sensitivity (64/83 = 77%) and specificity (1860/1902 = 98%). Of the identification panels, manual SPRCA identified more antibodies than PEG (67 versus 66) but had more inconclusive results (41 versus 20). Of overall strategies, manual SPRCA screening with either sequence of identification methods identified the most antibodies (66). The combination of PEG screen, PEG identification, and manual SPRCA identification identified slightly fewer antibodies (63) but had the lowest reagent cost, total (reagent plus labor) cost, and cost per antibody identified. The sequence of automated SPRCA screening with manual SPRCA identification, and PEG identification had the lowest hands-on time. CONCLUSIONS: The most cost-effective pretransfusion strategy is PEG screen with PEG identification, plus manual SPRCA identification when PEG identification is inconclusive.

Towards structural determination of the water-splitting enzyme. Purification, crystallization, and preliminary crystallographic studies of photosystem II from a thermophilic cyanobacterium.

A photosystem II preparation from the thermophilic cyanobacterium Synechococcus elongatus, which is especially suitable for three-dimensional crystallization in a fully active form was developed. The efficient purification method applied here yielded 10 mg of protein of a homogenous dimeric complex of about 500 kDa within 2 days. Detailed characterization of the preparation demonstrated a fully active electron transport chain from the manganese cluster to plastoquinone in the Q(B) binding site. The oxygen-evolving activity, 5000-6000 micromol of O(2)/(h.mg of chlorophyll), was the highest so far reported and is maintained even at temperatures as high as 50 degrees C. The crystals obtained by the vapor diffusion method diffracted to a resolution of 4.3 A. The space group was determined to be P2(1)2(1)2(1) with four photosystem II dimers per unit cell. Analysis of the redissolved crystals revealed that activity, supramolecular organization, and subunit composition were maintained during crystallization.

The diagnosis and significance of visceral pleural invasion in lung carcinoma. Histologic predictors and the role of elastic stains.

Invasion of the visceral pleura is an important component of lung carcinoma staging, and in some studies is an independent prognostic indicator. Evaluation of invasion by H&E may be indeterminate. Elastic stains can be helpful but are performed rarely. We reviewed all lung carcinoma resections from 1993 for 13 histologic features potentially predictive of pleural invasion. Of 57 resections, 20 were indeterminate by H&E. Verhoeff-Van Gieson (VVG) stain revealed invasion in 8 cases, increasing the pathologic stage in 1. VVG stain was negative in 12 cases, 2 of which had been falsely reported as positive, decreasing the stage in 1. Angiolymphatic invasion and single-cell spread were significant predictors of invasion. Absence of both or the presence of intervening aerated parenchyma predicted lack of involvement in all cases. Elastic stains can provide prognostically important information, changing the pathologic stage in 4% of lung carcinoma resections overall and in 10% of cases indeterminate by H&E for pleural invasion.

The generation of human dendritic and NK cells from hemopoietic progenitors induced by interleukin-15.

Interleukin-15 (IL-15) is a pleiotropic cytokine that induces the generation and differentiation of lymphoid cells and shares many biological activities with IL-2. We have shown here the development of dendritic cells (DC) from human CD34+ hemopoietic precursor cells cultured for 2-4 weeks with IL-15 alone. DC generated with IL-15 have typical morphological, immunocytochemical, phenotypic, and functional characteristics of mature DC. Dual flow cytometry analysis performed weekly demonstrated increasing co-expression of CD1a or CD83 with HLA-DR, CD80, CD86, IL-2R alpha, beta, and gamma. Two populations of cells were distinguished among CD34+ progeny. Small and medium-size cells were mainly natural killer (NK) cells (72.6-85.2% CD56+) and low numbers of DC (9.1-21.3% CD1a+). Large cells were mostly DC (75.4-95.4% CD1a+). Isolated CD34+ cells did not express IL-2R subunits but after 2-3 days in culture with IL-15, they were found to express IL-2Rgamma. Induced expression of IL-2Rgamma on CD34+ cells may explain the primary mechanism of IL-15-regulated differentiation of hemopoietic precursor cells. Thus, our data suggest that IL-15 stimulates CD34+ cells to differentiate into NK and DC and may represent a new growth and survival factor for lymphoid DC.

Interleukin-2-induces development of denditric cells from cord blood CD34+ cells.

Dendritic cells (DC) have been shown to develop along a myeloid or lymphoid lineage of differentiation propagated from bone marrow or early thymic precursor cells with hematopoietic cytokines. In our study, we have induced growth and differentiation of DC from cord blood CD34+ cells initiated in interleukin-2 (IL-2) alone or in IL-2 + stem cell factor (SCF) + tumor necrosis factor alpha (TNF-alpha)-supplemented medium and cultured with IL-2 or IL-2 + SCF for 28-35 days. Dendritic morphology and antigenic phenotype of DC grown with IL-2 were characteristic for DC cultured in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF). Growth and differentiation of DC was followed by an increase in expression of MHC II and co-stimulating molecules CD80 and CD86. We have also shown the expression of the IL-2 receptor (IL-2R) gamma-chain in CD34+ cells after 2-3 days of culture with IL-2 alone. The co-expression of the IL-2R alpha, beta, and gamma subunits in both DC cultured with IL-2- or GM-CSF-containing cocktail of cytokines was also shown. The time curve for induction of IL-2R demonstrated low levels of subunit expression at the beginning of culture. The number of CD1a cells co-expressing CD25, CD122, and CDgamma increased to about 24-68 and to 78-95% after 21 and 28-35 days, respectively. Development of natural killer cells was shown along with DC. The proportion of CD56+ cells and cytotoxicity increased in a time-dependent manner.

Attenuation of antipsychotic-induced hyperprolactinemia with clozapine.

Hyperprolactinemia is a well-known consequence of conventional antipsychotic therapy. The atypical antipsychotic clozapine is reported to lack this effect. We describe a case of attenuated serum prolactin levels after conversion to clozapine therapy in an adolescent. A 13-year-old female patient developed hyperprolactinemia with galactorrhea and amenorrhea while receiving thioridazine 300 mg daily. These symptoms continued throughout 3 years of treatment with haloperidol 10 mg daily and then fluphenazine 10 mg daily. Subsequently, after an incomplete improvement in her psychiatric symptoms and hyperprolactinemia on thioridazine 150 mg and bromocriptine 15 mg daily, the patient was changed to clozapine at age 16. Clozapine 150 mg twice daily improved her psychiatric status and corrected her serum prolactin concentrations after 2 weeks; bromocriptine was able to be discontinued. We recommend systematic evaluation of atypical neuroleptics as alternative treatments for refractory hyperprolactinemia induced by conventional antipsychotics.

Longitudinal analysis of abnormal involuntary movements in long-term clozapine-treated patients.

As part of a prospective efficacy and safety monitoring system, patients receiving clozapine are assessed monthly for dyskinetic events (DE), using the Abnormal involuntary Movement Scale (AIMS). Longitudinal analysis of 45 patients revealed 20 with baseline DE, 7 who developed emergent DE after a negative baseline assessment, and 18 patients with no DE symptoms throughout treatment with clozapine. Eight of the 20 patients with baseline DE were assessed to resolution of symptoms, with an average time of 261 +/- 188 days; 5 were evaluated until complete resolution of symptoms (AIMS = 0), with an average time of 691 +/- 462 days. The average time to onset of DE in emergent DE patients was 238 +/- 179 days, and the average time to resolution was 347 +/- 179 days after diagnosis. Four patients attained complete resolution with an average time of 629 +/- 293 days after diagnosis. It appears this emergent type of dyskinesia is different from other currently described dyskinesias. Overall, of the 27 patients having DE at any point in treatment, 15 of 27 (56%) had resolution of symptoms and 10 of 27 (37%) had complete resolution of DE. Clinicians should be aware of the utility of clozapine in dyskinesia and the extended time frame of response.

Acetylcholinesterase staining in the auditory brainstem nuclei of the leopard frog, Rana pipiens.

We examined the distribution of acetycholinesterase (AChE) in the brainstem auditory nuclei of leopard frogs (Rana pipiens). Numerous AChE-labeled somata were observed in caudal brainstem nuclei including the dorsolateral nucleus, superior olivary nucleus, and superficial reticular nucleus. At midbrain levels, AChE-labeled somata were observed in the magnocellular, principal and laminar nuclei of the torus semicircularis, though, most were located in the magnocellular nucleus. Neuropil labeling, while present throughout the auditory brainstem, was particularly pronounced in the principal nucleus of the torus. These findings suggest that the auditory system of anurans, like that of birds and mammals, comprises, in part, distinct cholinergic pathways.

Staphylococcus epidermidis bacteremia from transfusion of contaminated platelets: application of bacterial DNA analysis.

Septicemia is a rare complication of platelet transfusion. A case is reported of transfusion-associated septicemia in a 66-year-old man who received a 10-unit pool of platelets. During transfusion, he experienced rigors, wheezing, dyspnea, and fever. A total of four blood cultures drawn 10 and 36 hours after discontinuation of the transfusion grew Staphylococcus epidermidis. Culture of the residual platelet pool yielded S. epidermidis with a colony count of 10(5) organisms per mL. Strain identity of all four blood isolates and the platelet pool isolate was confirmed by gel electrophoresis of EcoRI and HindIII restriction digests of whole-cell DNA. There have been 31 prior reported cases of platelet transfusion-associated septicemia, of which 9 have been caused by coagulase-negative staphylococci. Systemic reactions to platelet transfusions should prompt consideration of transfusion-associated bacteremia as the cause.

Warthin's tumor with malignant lymphoma. DNA analysis of paraffin-embedded tissue.

The authors report a case of malignant lymphoma, small lymphocytic type, involving the lymphoid stroma of a Warthin's tumor of the parotid gland. This was confirmed by the presence of a monoclonal immunoglobulin heavy chain gene rearrangement, demonstrated by Southern blot hybridization of DNA extracted from paraffin-embedded tissue. Similar techniques showed only germline immunoglobulin gene bands in two control cases of Warthin's tumor.

Dermatophilus congolensis and "hairy" leukoplakia.

The authors report the first human case (to our knowledge) of infection of the oral mucosa by Dermatophilus congolensis. Septate branching filaments morphologically identical to those of D. congolensis were identified in the lingual epithelium of a male homosexual employed as an animal handler. This actinomycete is the cause of dermatophilosis, a proliferative exudative dermatitis affecting many animal species. Clinical features suggested "hairy" leukoplakia (HL), a hyperkeratotic tongue lesion for which human papillomavirus (HPV) and Epstein-Barr virus (EBV) have been implicated as etiologic agents. Immunoperoxidase staining for HPV capsid antigen was negative. Direct immunofluorescent staining with a conjugate specific for D. congolensis identified the bacterial structures as those of this species while excluding morphologically similar organisms.

Caffeine content of common beverages.

Tea, coffee, carbonated and chocolate beverages were analyzed for caffeine, and results compared in terms of usual serving sizes. Significant differences in caffeine levels were found to result from the preparation method of coffee or brewing time of tea. It is possible for a cup of tea, instant coffee, or can of cola beverage to have similar caffeine content (55 to 65 mg.); however, the mean values per cup of black tea (28 to 46 mg.) are considerably lower than for brewed coffee (107 to 151 mg.). Caffeine is readily absorbed and can have pharmacologic effects on adults or on children who consume quantities of cola beverages or chocolate. Both preparation method and quantity of beverage consumed should be considered in taking dietary histories or estimating caffeine intake.

Preventive effect of estrogen on postmenopausal bone loss.

Follow-up studies of bone mineral content in the radius were done in 82 postmenopausal women 4 to 10 years after the first examination. These patients were subdivided into four groups depending on the type of menopause (artificial or natural) and estrogen administration (treated or untreated). Bone mineral mass and combined cortical thickness decreased significantly in both groups of untreated women. Both mineral loss per year for the untreated women was -9.1 mg/sq cm for castrates and -6.9 mg/sq cm for those with a natural menopause. In neither group was the rate of loss correlated with age. The change in bone mineral mass per year in the estrogen-treated subjects (mean +3.25 mg/sq cm) differed significantly from that of untreated subjects (mean -7.99 mg/sq cm). The findings suggest that postmenopausal osteoporosis could be prevented by estrogen treatment.

Variability of creatinine excretion of normal, phenylketonuric and galactosemic children, and children treated with anticonvulsant drugs.

Creatinine per minute excretion rates in individual voidings varied as much as 300-500 percent of the 24-hour value for several children. Creatinine excretion rates were significantly more variable for children treated with anticonvulsant drugs than for normal children. Riboflavin-creatinine ratios determined on individual voidings were variable but adequate although total riboflavin was low. No time of day was found when creatinine excretion was representative of the 24-hour value. The creatinine height index when calculated from timed individual voidings during a 24-hour period varied for some children from 0.5 to more than 1.5 times published norms.