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Molecular typing of Plasmodium falciparum from Giemsa-stained blood smears confirms nosocomial malaria transmission.

In this report, we describe the partial molecular characterisation of Plasmodium falciparum isolates obtained from two individuals who were involved in a probable case of accidental malaria transmission after admission to a hospital in the metropolitan area of São Paulo, Brazil. Molecular analysis of polymorphic stretches of the merozoite surface protein 1 and 2 genes using PCR-typing and nucleotide sequencing revealed that the two isolates were identical and that the identified msp-1 gene was different from all others published to date. Additional anamnestic data supported our findings and made all other possible routes of infection unlikely. The methodology used here is simple to perform and needs as little as one Giemsa-stained blood smear as starting material.

Adult↗

Comparison of nuclear shape in aspirated and histologic specimens of prostatic carcinoma.

Nuclear shape analysis of histologic sections of operative specimens has separated patients with clinically localized prostatic carcinoma with good and poor prognoses. In order to become more useful clinically, nuclear shape should be evaluable preoperatively but is distorted by core biopsy. We used nuclear morphometry to compare histologic and cytologic specimens obtained from 20 patients who underwent radical prostatectomy for clinically localized prostatic carcinoma. Neoplastic nuclei from cytologic aspirates prepared by the Papanicolaou and Diff-Quik techniques and standard histologic sections stained with hematoxylin and eosin were digitized. Air-dried nuclei of the Diff-Quik preparation had a nuclear area of 104.2 microns 2 +/- 28.2 SD. These nuclei were nearly twice as large (paired t test, P < .001) as in the Papanicolaou (55.6 +/- 13.0 microns 2) and histologic (55.8 +/- 12.7 microns 2) preparations. However, nuclear shape was not affected. Nuclear roundness factor (deviation from circularity) and boundary curvature were similar (paired t tests, P > .05). All nuclear shape descriptors of cytologic smears fell within intraobserver variations of measurements in histologic sections. Nuclear shape is not altered in Papanicolaou- and Diff-Quik-stained cytologic specimens and should be tested for preoperative assessment of outcome in clinically localized prostatic carcinoma.

Aged↗

Assessment of neonatal conjunctivitis with a direct immunofluorescent monoclonal antibody stain for Chlamydia.

Chlamydial conjunctivitis was diagnosed by direct immunofluorescent monoclonal antibody staining of conjunctival smears in 46 of 100 consecutive neonates with conjunctivitis. The remaining 54 infants had bacterial conjunctivitis most commonly caused by Staphylococcus, Streptococcus, and Haemophilus species. A comparison of the direct test with chlamydial cultures demonstrated a sensitivity of 100% and a specificity of 94%, whereas Giemsa stain had a sensitivity of 42% and a specificity of 98%. Nineteen percent of infants treated with oral erythromycin for chlamydial infections in accordance with Centers for Disease Control guidelines had clinical and laboratory evidence of persistent chlamydial conjunctivitis. We conclude that Chlamydia trachomatis is a major cause of neonatal conjunctivitis that can be effectively and rapidly diagnosed by direct immunofluorescent monoclonal antibody staining of conjunctival smears. Further evaluation of treatment regimens appears to be warranted.

Antibodies, Bacterial↗

[An unusual heteromorphic bivalent in the common vole (Microtus arvalis) from Byelarus].

Electron microscopic analysis was carried out on the synaptonemal complexes of ten male common voles (Microtus arvalis) caught of 1990 in Byelorussia. In the early pachytene stage of spermatocytes of four males heteromorphic bivalent has been found in one of five large autosomes. In the central region of the bivalent one of the lateral elements is in the form of a D-loop, characteristic of insertion/deletion heterozygotes. However, high-resolution G-band staining of mitotic chromosomes from fibroblasts showed no significant differences in the G-band patterns between homologues.

Animals↗

Helicobacter pylori in gastric biopsy specimens. Comparison of culture, modified giemsa stain, and immunohistochemistry. A retrospective study.

Antral biopsy specimens of 302 different endoscopic investigations of 200 patients with non-ulcer dyspepsia were studied for the presence of Helicobacter pylori in order to determine the most sensitive detection method. Part of the biopsy was cultured, and part stained using a modification of the Giemsa stain, and with an immunoperoxidase technique using a polyclonal rabbit anti-H. pylori antiserum. Cross-reactivity of this antiserum with other Campylobacter species was minimal. Material from 244 investigations was studied using all three detection methods. Culture was positive in 44 per cent, Giemsa in 78 per cent, and immunoperoxidase in 89 per cent of these biopsy specimens. Only five positive Giemsa stains with negative immunoperoxidase stain were found, whereas in 32 cases a negative Giemsa stain with a positive immunoperoxidase stain was seen. In the latter cases, the bacterial load was very low. The specimens revealed bacteria only sporadically, always confined to the deep layers of the gastric pits. Culture results correlated significantly with the bacterial load observed in the Giemsa stain. It is concluded that culture of H. pylori is the least sensitive detection method, whereas immunoperoxidase staining is the most sensitive. For daily practice the modified Giemsa stain, however, appears to be sufficient to diagnose the presence of the micro-organism.

Adolescent↗

G-banding and chromosome condensation in the ant, Tapinoma nigerrimum.

Well-defined G-bands were obtained on metaphase chromosomes from Tapinoma nigerrimum using trypsin and warm 2 x SSC in sequence. The G-banded pattern allowed the identification of all chromosomes. Evidence for asynchronous condensation of the chromosomes of this species is provided. Different banding patterns were obtained when metaphase chromosomes were stained with DA/DAPI alone and with DA/DAPI after a standard G-banding procedure. The G-banding phenomenon is discussed using the result obtained.

Animals↗

Chromosomal aberrations induced in human lymphocytes by high-LET irradiation.

High linear energy transfer (LET) particles are more efficient than sparsely ionizing radiations in inducing chromosomal aberrations, in particular complex rearrangements. We analysed R-banded chromosome rearrangements in human lymphocytes irradiated with several ions having a wide range of LET (31.3-1435 keV/micron). The frequency of chromosome breaks unrejoined or inferred from observed rearrangements, and of complex rearrangements induced by a single particle, increased with the LET up to about 100-150 keV/micron and seemed to level off for higher LET values. Additional study was focused on damage induced by oxygen ions of three different energies. Significant cell cycle delay, and multiple chromosome rearrangements and breaks were demonstrated using Giemsa and Fluorescence-plus-Giemsa stainings, coupled with chromosome painting. Damage increased with the fluence and the LET, but at the higher LET damage decreased for fluences > 10(7) particles/cm2. Cell death and G2 block might be involved in this phenomenon. Chromosome 1 painting exhibited a high frequency of breaks and complex rearrangements, which would not have been detected using a standard staining. Complex rearrangements were induced by as few as one particle per cell nucleus and may be considered as a biological fingerprint of high-LET irradiation.

Azure Stains↗

A carmine-Giemsa staining technic for meiotic prophase chromosomes of the genus Beta L.

For a detailed study of chromosome morphology in meiotic prophase stages of Beta species, a special double staining technic has been developed. It consists of combined maceration-staining in an ethanol-hydrochloric acid-carmine mixture followed by poststaining of the squashed material in a diluted Giemsa solution. The technic yields well-spread prophase meiotic nuclei showing detailed structures both in weaker stained chromosome segments and in threadlike chromatin structures. This technic proved to be especially favorable for stages which are difficult to interpret, such as pachytene, schizotene and diffuse stages.

Azure Stains↗

Wuchereria bancrofti: the staining of the microfilarial sheath in giemsa and haematoxylin for diagnosis.

Indigenous cases of bancroftian filariasis have recently been identified in the country for the first time. The diagnosis of such infections depends on demonstrating the characteristic sheathed microfilariae of Wuchereria bancrofti in the blood. Giemsa, a stain recommended by many authors, was found to be unsuitable as the microfilarial sheath, an important distinguishing feature, does not take up the stain. In contrast, the sheath stains clearly in haematoxylin. The differences in the staining reaction of the microfilariae in the two stains is demonstrated. The need to establish the presence of the sheath is emphasized, especially since the microfilariae of Dipetalonema perstans, the widespread filarial infection in the country, are unsheathed.

Azure Stains↗

47,X,i(Xq),Y karyotype in Klinefelter's syndrome.

This is a case report of 47,X,i(Xq),Y in a 24-year-old infertile male with Klinefelter's syndrome. C staining indicated that this isochromosome X had a single small centromere. BUdR incorporation revealed the isochromosome X to be late replicating.

Azure Stains↗

Cytochemical heterogeneity of the C-band in human chromosome 1.

The heterogeneity of the C-band of human chromosome 1 has been evaluated using several selective staining methods: C-banding (CBG), distamycin A plus 4-6-diamidino-2-phenylindole (DA/DAPI) and Giemsa G-11 pattern following the treatment with the restriction endonucleases AluI and HaeIII. The bands produced by each method are characteristic but not identical. The total C-band is resistant to AluI treatment. The bands induced by HaeIII and the one stained by DA/DAPI are markedly similar but smaller than the C-band. The G-11 technique stains yet smaller regions than those of HaeIII and DA/DAPI. Depending on the expression of staining properties, the C-band of chromosome 1 usually consists of three subdivisions: the proximal, intermediate and distal regions, suggesting an extremely heterogeneous nature. The staining variations between different regions are further substantiated by studies of a reciprocal translocation where the proximal region and the remaining C-band of chromosome 1 are separate.

Azure Stains↗

Diagnosis of malaria by acridine orange fluorescent microscopy in an endemic area of venezuela.

Fluorescent (acridine orange) microscopical examination of capillary centrifuged blood (quantitative buffy coat [QBC] analysis) and Giemsa stained thick blood smears (GTS) were compared for diagnosis of malaria in blood specimens from adults living in malaria transmission areas of the States of Bolivar and Amazonas in southeastern and south Venezuela, respectively. Of a total of 198 GTS examined, 95 subjects (48%) showed parasitaemia. Among the 95 blood films with a positive GTS, 94 were judged positive by the QBC. However, positive QBC tubes were found in 29 out of 103 blood specimens with a negative GTS. Thus, relative to a GTS standard, the sensitivity and specificity of the QBC-test was 99.2% and 72%, respectively. Young trophozoites of Plasmodium vivax and P. falciparum could not be distinguished with certainty. It is confirmed that the QBC offers many advantages compared with the standard diagnosis of malaria parasites, specifically in the speed of staining and ease of interpretation. However, in places where P. falciparum and P. vivax occur, species and stage differentiation should be confirmed with the GTS.

Acridine Orange↗

Quick and reversible staining methods for G- and Q-bands of chromosomes.

Staining G-bands of chromosomes can be carried out in 2 1/2 minutes using a mixture of Giemsa solution with a special diluting preparation. The staining is completely reversible without any damage to the chromosomes. Moreover, an improved method for staining Q-bands with atebrin and mounting the preparation in sucrose is described. The whole process is performed in one minute and the preparations keep for weeks at laboratory temperature.

Azure Stains↗

Cytology of polychrome-stained equine synovial fluid smears. Comparison with clinical findings, histologic specimens, Wright-Giemsa-stained smears and outcome.

Polychrome-stained equine synovial fluid specimens from 34 normal joints and 129 joints with clinical abnormalities were examined cytologically. The smears from joints with abnormalities were categorized as within normal limits (4.7%), slight abnormality (27.9%), proliferative synovitis (21.7%), neutrophilic pattern (20.2%), elongated cell pattern (10.1%), other moderate to marked abnormality (11.6%) and unsatisfactory (3.9%). Cytologic abnormalities that were not restricted to a single category included spindle cells, crystals, stellate cells and cartilage fragments. Multinucleate cells and mononucleate cells with dense cytoplasm and a delicate periphery were seen in smears from cases with clinical diagnoses of osteochondrosis or fracture; interpretation of these cells as osteoclasts and their mononucleate precursors was supported by positive staining with tartrate-resistant acid phosphatase. Smears within the same cytologic category were not found to correspond with a single clinical diagnosis. The identification of several cytologic patterns in cases with the same clinical diagnosis suggests that multiple stages of disease were sampled. Except in cases with the cytologic neutrophilic pattern, there was not a consistent relationship between the histologic features in synovial biopsy specimens and the cytologic findings; the morphologic variation within synovial membrane sections and between sections from different locations was sometimes marked. When compared with air-dried, Wright-Giemsa-stained smears, the polychrome-stained smears were more sensitive in the detection of cytologic abnormalities and were less often falsely negative or unsatisfactory. Following surgery, cases with clinical diagnoses of osteochondrosis (29 cases) and fracture (25 cases) were analyzed according to clinical outcome and cytologic category. While 80% of the horses with proliferative synovitis in cytologic specimens were sound, only 67% of those with the elongated cell pattern, 50% of those with slight abnormality and 33% of those with other moderate to marked abnormality were sound. A statistically significant relationship (P less than .02) was found in cases with a diagnosis of osteochondrosis: animals with a proliferative synovitis pattern were almost three times as likely to be sound as compared to those with slight abnormality. These findings indicate that polychrome-stained equine synovial fluid smears (1) provide information that is different from that found in corresponding histologic sections and (2) are superior to air-dried, Wright-Giemsa-stained smears for cytologic examination. The polychrome-stained equine synovial fluid smears were found to provide information supportive of clinical, radiographic and prognostic data.

Animals↗

Evolution of human cytogenetics: an encyclopedic essay. III. The second decade after 1956: banding techiques.

Unequivocal establishment of the correct diploid chromosome number in 1956 started the modern era of human cytogenetics. The next impetus came when the peripheral blood leukocyte culture technique for the chromosome preparation was described in 1960. Discovery of special staining procedures - banding techniques - in early seventies not only saved it from early senescence but played decisive roles in broadening the horizons of modern human cytogenetics.

Acridines↗

R-banding produced by DNase I digestion of chromomycin-stained chromosomes.

A distinct reverse (R-) banding pattern was produced on human chromosomes by digesting chromosome spreads with pancreatic deoxyribonuclease I (DNase I) in the presence of an excess of chromomycin A3 (CMA), followed by staining with Giemsa. The banding pattern corresponds with that obtained by chromomycin A3 fluorescence, and bands which fluorescence brightly with chromomycin appear darkly with Giemsa. The same relationship was observed in two plants, Scilla siberica and Ornithogalum caudatum, which have contrasting types of heterochromatin. Chromomycin bright C-bands stained darkly with the CMA/DNase I technique, whereas chromomycin negative C-bands appeared lightly stained. The digestion patterns are thought to reflect the variation in chromomycin binding capacity along the chromosome with R-bands and dark C-bands being sites which preferentially bind the antibiotic.

Azure Stains↗

Squamous cell carcinoma of the conjunctiva. Value of exfoliative cytology in diagnosis.

Cytologic examination is of potential value to the practicing ophthalmologist in confirming the clinical impression of cancer of the eye. Scrapings of conjunctival lesions can be prepared in the office, stained with Giemsa stain and rapidly examined. Carcinoma in situ and squamous cell carcinoma of the bulbar conjunctiva were presented as illustrations of the usefulness of exfoliative cytology in clinical ophthalmology.

Adult↗