Evaluation of the Microsemi CRP, an automated hematology analyzer for rapid 3-part WBC differential and CRP using whole blood.

INTRODUCTION: We evaluated the basic performance of Microsemi CRP, an unique automated hematology analyzer which can simultaneously measure CBC including 3-part WBC differential (3-Diff) and CRP using whole blood treated with EDTA-2K anticoagulant. METHOD: We found that it produced generally the acceptable results for all parameters performed (repeatability, reproducibility, linearity, interference effect, carry over, and correlation) using control materials, fresh human whole bloods, and serum samples. RESULTS: CBC data examined using Microsemi CRP showed the good correlation with the previous model, Micros CRP200 (r ≧ 0.9), and also those obtained using the routine analyzer, ADVIA 2120i (r ≧ 0.989). Concerning the 3-Diff, both GRA (%) and LYM (%) showed the excellent correlation coefficient between Microsemi CRP and Micros CRP200 (r ≧ 0.992) as well as ADVIA 2120i (r ≧ 0.957). MON (%) showed good correlation between Microsemi CRP and Micros CRP200 (r = 0.959), but lower correlation between Microsemi CRP and ADVIA 2120 i (r = 0.471). CRP data showed the good correlation with HITACHI7600 (r ≧ 0.997) and Micros CRP200 (r ≧ 0.997). CONCLUSION: From these findings, we concluded that Microsemi CRP seemed the convenient laboratory analyzer in the setting of point of care testing (POCT) especially at NICU or primary care unit.

Self-monitoring of home blood pressure with estimation of daily salt intake using a new electrical device.

We investigated a simple device to monitor daily salt intake at home and examined the relationship between salt excretion and morning blood pressure in order to enable patients to better manage daily salt intake and hypertension. The correlation between 24-h urinary salt excretion and measured value with salt monitor from overnight urine was significant (n = 224, r = 0.72, P < 0.001). A total of 46 volunteers participated for more than 3 weeks by measuring daily salt intake and morning blood pressure. The relationship between predicted daily salt excretion and blood pressure was examined with use of 3-day moving average. Mean salt excretion and systolic blood pressure (SBP) significantly decreased by the end of the trial (i.e., salt excretion decreased from 158+/-31 to 149+/-30 mmol/day and SBP from 137+/-17 to 133+/-16 mm Hg). Of 46 participants, 18 (39%) had a significant correlation between predicted daily salt excretion and blood pressure (r > 0.4, P < 0.05, n > 21), indicating sodium sensitivity. An additional 17% had a positive correlation that did not reach statistical significance (0.2 < r < or = 0.4), and the remaining 44% had no correlation (r < or = 0.2). Mean decrease in blood pressure per decrease in salt (g) (17 mmol) intake in the 18 participants with a significant correlation was 3.3 mm Hg (SBP) and 1.5 mm Hg (diastolic blood pressure), which was higher than that reported for other studies. Hypertensive patients not using medication showed the largest decrease. We conclude that daily monitoring of salt intake and morning blood pressure will be useful for management of hypertension.

Prevalence of Cryptosporidium parvum infection and pattern of oocyst shedding in calves in Japan.

Cryptosporidium parvum infection and the pattern of oocyst shedding were observed in calves. A total of 480 fecal samples were collected from 30 calves (age, < or =30 days) over a period of 10 months from June 1998 to March 1999. A sucrose centrifugal flotation technique revealed 28/30 (93%) calves were passing Cryptosporidium oocysts. Oocyst shedding was first detected on the sixth day after birth, with 8% of the calves testing positive. This rate increased day by day and reached approximately 80% by day 15. Oocyst shedding varied from 1 to 13 days, with a mean of 7 days. Calves infected with C. parvum had a significantly higher rate of diarrhea (33%) than non-infected calves (8%) (P<0.05), suggesting C. parvum infection as the likely cause. The mean number of oocysts excreted by calves < or =30 days old was approximately 6x10(7) per gram of feces. These results indicated that one calf would excrete some 6x10(11) oocysts in the first month after birth, taking both the quantity of feces in a day and the period of excretion into consideration. Accordingly, it is clear that calves are important in the spread of cryptosporidiosis to calves and humans.

Mapping of interneurons that contribute to food aversive conditioning in the slug brain.

To determine the distribution of neurons that contribute to memory formation induced by odor-taste associative conditioning in the slug's brain, we examined neuronal activity of the central nervous system of the slug Limax marginatus using a fluorescent activity marker [Lucifer yellow (LY)]. When LY was injected into the body cavity just after the conditioning, many of the procerebral (PC) interneurons were labeled. The PC lobe was considered to play important roles in the olfaction of the slug, because the olfactory afferent fibers from both the inferior and the superior tentacular noses innervate it. Such strong dye-uptake activity of PC interneurons was not observed when LY was injected just after unpaired control treatment. Thus, it was suggested that enhancement of dye-uptake activity upon conditioning was caused by the association of a conditioning stimulus (CS) with an unconditioned stimulus (UCS). The distribution patterns of PC interneurons that were labeled by LY after conditioning showed a characteristic feature: They usually formed a belt-shaped cluster parallel to the dorsoventral axis. This feature of the distribution was maintained when different odors were used as a CS. Furthermore, the number of the clusters reflected the number of CS odors but not the number of conditioning sessions. From these observations, we considered that enhancement of the neural activity involving dye uptake in each belt-shaped cluster contributed to formation of each odor memory.

Eruptive clear cell hamartoma of sweat duct.

A histologically unique condition of the sweat duct was found in a miliaria-like papular eruption on the extensor surfaces of extremities of a 41-year-old female patient. The hypertrophic epithelium of the sweat duct wall consisted of extensively vacuolated clear cells. There was mild hyperkeratosis around the sweat duct orifice. The eccrine gland cells showed only partial vacuolization. Since this condition is not related to any of the other known benign changes of the eccrine duct, we report it with a presumptive descriptive diagnosis of eruptive clear cell hamartoma of sweat duct.

Time-resolved electron microscopic analysis of the behavior of myosin heads on actin filaments after photolysis of caged ATP.

The interaction between myosin subfragment 1 (S1) and actin filaments after the photolysis of P3-1-(2-nitrophenyl)ethyl ester of ATP (caged ATP) was analyzed with a newly developed freezing system using liquid helium. Actin and S1 (100 microM each) formed a ropelike double-helix characteristic of rigor in the presence of 5 mM caged ATP at room temperature. At 15 ms after photolysis, the ropelike double helix was partially disintegrated. The number of S1 attached to actin filaments gradually decreased up to 35 ms after photolysis, and no more changes were detected from 35 to 200 ms. After depletion of ATP, the ropelike double helix was reformed. Taking recent analyses of actomyosin kinetics into consideration, we concluded that most S1 observed on actin filaments at 35-200 ms are so called "weakly bound S1" (S1.ATP or S1.ADP.Pi) and that the weakly bound S1 under a rapid association-dissociation equilibrium with actin filaments can be captured by electron microscopy by means of our newly developed freezing system. This enabled us to directly compare the conformation of weakly and strongly bound S1. Within the resolution of deep-etch replica technique, there were no significant conformational differences between weakly and strongly bound S1, and neither types of S1 showed any positive cooperativity in their binding to actin filaments. Close comparison revealed that the weakly and strongly bound S1 have different angles of attachment to actin filaments. As compared to strongly bound S1, weakly bound S1 showed a significantly broader distribution of attachment angles. These results are discussed with special reference to the molecular mechanism of acto-myosin interaction in the presence of ATP.

Elastic filaments in situ in cardiac muscle: deep-etch replica analysis in combination with selective removal of actin and myosin filaments.

To clarify the full picture of the connectin (titin) filament network in situ, we selectively removed actin and myosin filaments from cardiac muscle fibers by gelsolin and potassium acetate treatment, respectively, and observed the residual elastic filament network by deep-etch replica electron microscopy. In the A bands, elastic filaments of uniform diameter (6-7 nm) projecting from the M line ran parallel, and extended into the I bands. At the junction line in the I bands, which may correspond to the N2 line in skeletal muscle, individual elastic filaments branched into two or more thinner strands, which repeatedly joined and branched to reach the Z line. Considering that cardiac muscle lacks nebulin, it is very likely that these elastic filaments were composed predominantly of connectin molecules; indeed, anti-connectin monoclonal antibody specifically stained these elastic filaments. Further, striations of approximately 4 nm, characteristic of isolated connectin molecules, were also observed in the elastic filaments. Taking recent analyses of the structure of isolated connectin molecules into consideration, we concluded that individual connectin molecules stretched between the M and Z lines and that each elastic filament consisted of laterally-associated connectin molecules. Close comparison of these images with the replica images of intact and S1-decorated sarcomeres led us to conclude that, in intact sarcomeres, the elastic filaments were laterally associated with myosin and actin filaments in the A and I bands, respectively. Interestingly, it was shown that the elastic property of connectin filaments was not restricted by their lateral association with actin filaments in intact sarcomeres. Finally, we have proposed a new structural model of the cardiac muscle sarcomere that includes connectin filaments.

Density values of the corpus callosum in schizophrenic brains.

Density values of the corpus callosum and ventricular brain ratio (VBR) were obtained by coronal computerized tomography (CT) in 16 chronic schizophrenic patients and 16 normal controls. The corpus callosum of schizophrenic brains showed no apparent density loss which could indicate a dysfunctional brain. A significant ventricular dilatation was found in the patient group compared with age-matched controls. This finding was compatible with several former observations with transverse scanning.

Lesions of the mediodorsal thalamic nucleus do not change thresholds for hypothalamic defensive attack in cats.

Electrolytic lesions of the mediodorsal thalamic nucleus (nMD) were made in 10 cats to evaluate their effects on thresholds for hypothalamic defensive attack and emotional reactivity to noxious stimuli. Only 2 cats showed minimal threshold changes after the lesions, one of which was accompanied by an increased emotional reactivity to noxious stimuli; however, this was attributed to damage to the fornical column. The nMD appeared not to be involved directly in the central mechanism of defensive attack.

Computerized tomographies of 34 patients at the chronic stage of acute carbon monoxide poisoning.

The brains of 34 patients at the chronic stage of acute carbon monoxide poisoning (CO poisoning) were examined using computerized tomography (CT). Ventricular and sulcal dilatations were measured quantitatively, with picture analysis of CT for the measurement of ventricular dilatation. Significant ventricular and sulcal dilatations were found in all cases of the CO group compared with age-matched controls, and bilateral low density areas in the globus pallidus were seen in 9 of the patients. There were significant correlations between duration of initial unconsciousness and the ventricular dilatation or cortical atrophy. Such dilatations were considered to be due to the cerebral damage in the acute stage.

Serological differentiation of strains of group B streptococci by the soft-agar technique.

Soft-agar technique was applied for serological determination of strains of group B streptococci. With representative type strains, their colonial morphology was converted from diffuse- to compact-type growth only by the addition of homologous rabbit anti-serum in the medium and no conversion of colonial morphology was observed by heterologous rabbit antisera. Twenty seven out of 30 fresh isolates obtained from human clinical specimens showed diffuse-type growth in soft-agar medium and were subjected to this examination. Twenty-one strains reacted with a single antiserum, 5 strains showed reactivities to two different antisera although reaction to an antiserum were significantly higher than those of the other antiserum and no reaction was observed with a strain. Twenty single and 5 major serotypes determined by this technique were coincided with those differentiated by Lancefield's precipitin method. From these experimental results, soft-agar technique was regarded being available for serological typing of strains of group B streptococci.

Comparison of three blood-clotting substances in Staphylococcus aureus strains.

Besides the two well-known blood-clotting substances, coagulase and clumping factor, a third one has been identified from the staphylococci which is a cell surface polysaccharide, is alkali stable, and induces compact-colony formation in serum soft agar. Using some 97 clinical strains of Staphylococcus aureus, we found that in production and activity the substances were distinctly different.

Relation of colonial morphologies of strain of Streptococcus pneumoniae in soft-agar to the encapsulation.

Among 40 fresh isolates of Streptococcus pneumoniae 12, 25 and 3 strains, respectively, exhibited large round, small round and compact colonial morphologies in soft-agar medium. Every large round strain possessed a capsule, almost half of the small round strains had capsules, while all of the large round type growth showed very high mouse virulence and 1.0 mg of these organisms was capable of absorbing a minimal amount of passive protective antibody in rabbit antiserum, prepared with the homologous strain, against challenge infection with homologous organisms in mice. Its variant showing compact type growth in soft-agar was mouse avirulent and a similar amount of the mouse passive protective antibody could not be absorbed with 100 mg of these organisms. These experimental results indicate that the soft-agar technique can be used for the identification of encapsulated strains of Streptococcus pneumoniae.

Passive protection by human serum in mice infected with encapsulated Staphylococcus aureus.

The occurrence and nature of passive protective antibody in 100 samples of human serum was investigated in mice challenged with strains of Staphylococcus aureus capsular types A (Smith diffuse strain) and B (strain NS58D). Sixty of the sera passively protected mice against the capsular type-A strain, three against type B, and one against both types. Rabbit antisera against human IgG, IgA and IgM could remove the protective activity from a human serum of high potency, and the activity was also sensitive to 2-mercaptoethanol. Absorption with Smith surface antigen removed protective activity and reduced the concentration of IgG 7-fold, IgA 2.7 fold and of IgM 3-fold more than in a non-protective serum. Consequently, the protective activity of human serum is believed to be associated with antibodies to the S. aureus capsular antigen in the three immunoglobulin classes.

Electronmicroscopic finding of surface structure and capsular swelling reaction of a strain of Klebsiella pneumoniae.

To investigate a capsular swelling reaction of the strain K-9 of Klebsiella pneumoniae, possessor of large capsule, ultra-thin sections of the organisms were stained with uranyl acetate and lead citrate and were treated with rabbit antiserum. The ultra-thin sections of above stained organisms showed spike-like structure in outmost layer of the cell wall. When they were treated with rabbit antiserum, the outmost layers of the organisms was surrounded by a zone of oval to round polymorphous vesicular structure which covered the spike-like appendages. These findings were assumed being capsular swelling reaction of these organisms. However, when they were treated with rabbit antiserum which was previously absorbed with the polysaccharide surface antigen, polymorphous vesicular structures did not appear and sharp spiked structures issuing from the cell wall were exposed.