Luis Alberto Palaoro

Utilidad de la técnica de AgNOR en la interpretación de los derrames de cavidades serosas

Background: AgNOR technique detects, using silver salts, argyrophylic proteins of the nucleolar organizer region (NOR). The number and size of NOR reflect cell activity, proliferation and transformation and may help to differentiate benign from malignant cells. Aim: To assess the value of AgNOR assay to differentiate reactive mesothelial cells from malignant cells in serous effusions. Material and methods: Thirty one fluids obtained from 16 pleural, 14 peritoneal and one pericardial effusion, were studied. The fluids were processed with Giemsa and Papanicolau stains and with the AgNOR technique. The number of AgNOR dots were counted (only when it was possible to distinguish each individual dot) and the mean value per nucleus was calculated for each smear. Results: Mesothelial cells had a mean of 4,88 ± 1,5 dots compared with 13,78 ± 3,88 dots in the malignant cells (p<0,001). Conclusions: AgNOR assay can be useful for the differentiation of benign and malignant cells in serous effusions. (Rev Med Chile 2000; 128: 963-8)

In vivo antigenotoxic activity of watercress juice (Nasturtium officinale) against induced DNA damage.

The present study was carried out to investigate the genotoxicity as well as possible protective activity against damage induced by cyclophosphamide (CP) of the aqueous juice of watercress (Nasturtium officinale, W.T. Aiton) in vivo. Male and female Swiss mice 7–8 weeks old (N = 48) were treated by gavage with 1 g kg−1 body weight and 0.5 g kg−1 body weight of watercress juice during 15 consecutive days. Genotoxicity and its possible protective effect were tested by the comet assay in peripheral blood cells and the micronucleus test in bone marrow. In addition, biopsies of the bladder, epididymis and testicles of mice were performed to extend the experimental design. Watercress juice per se did not induce genetic damage according to the comet assay and micronucleus study, exhibiting a protective activity against CP (P < 0.05 and P < 0.001, respectively). The comparative analysis of bladder histological changes obtained in the watercress plus CP group against those treated with CP alone suggests a probable protective effect. Further studies are needed in order to establish the protective role of watercress juice against DNA damage. Copyright

New Biochemical Parameters in the Differential Diagnosis of Ascitic Fluids

Background In the cases of ascitis, it is essential to determine their origin using the parameters obtained by the cytological and biochemical examinations. The aim of this study was to evaluate the usefulness of different biochemical markers and the number of cells in the differential diagnosis of ascitic fluid (AF). Methods One hundred ninety-one cases of AF were studied, who were admitted to the hospital from January 01, 2009 to December 31, 2014. One hundred fifty-two of them were included in the analysis, and the remaining 39 were excluded because they had more than one associated pathology, clotted or hemolyzed. Results The more frequent etiologies of AF were the cirrhosis (29%), the infections (22%) and the neoplasies (19%). Other pathologies reached 16%. Cutoff > 300 cells/mm3 detected the 78% of exudates. The AF/serum (S) of aspartate aminotransferase (AST) (> 0.5), lactate dehydrogenase (LDH) (> 0.6), proteins (PT) (> 0.5), cholesterol (COL) (> 0.4), and alanine aminotransferase (ALT) (> 0.5) correctly detected 80%, 78%, 72%, 70% and 70% of the exudates, respectively. Conclusion We proposed the utilization of a new cutoff of cellular counting, major of 300/mm3, since it would allow improving the detection of exudate ascites, without including the transudate ascites. AST AF/serum ratio (AF/S) showed the major usefulness in the differentiation and characterization of AF; LDH, proteins, cholesterol and ALT might be also acceptable in the above mentioned differentiation. The serum-ascites albumin gradient (SAAG) turned out to be a good marker of portal hypertension associated with cirrhotic processes. Creatine kinase (CK), alkaline phosphatase (ALP), amylase (AMI), total bilirubin (TB), triglycerides (TG) and glucose (GLU) did not allow differentiating exudates from transudates.

A rare case of respiratory disorders associated with two autosomal recessive diseases and male infertility.

The study of nasal ciliary beat frequency (CBF) and ultrastructure may contribute to the understanding of pathognomonic cases of male infertility associated with defects in sperm motility. This study was designed to report a particular case of male infertility, characterized by the association of two respiratory autosomal recessive genetic diseases (alpha-1-antitrypsin deficiency [AAT-D] and primary ciliary dyskinesia [PCD]). A 39-year-old patient with complete sperm immotility, AAT-D, and bronchiectasis was studied in the Laboratory of Male Fertility, the Department of Urology, the Respiratory Center of a Pediatric Hospital, and in the Department of Clinical Medicine of a Rehabilitation Respiratory Hospital. Family history, physical examination, hormonal analysis, microbial assays, semen analysis, nasal ciliary function, and structure study by digital high-speed video photography and transmission electron microscopy are described. A noninvasive nasal biopsy to retrieve ciliated epithelium lining the inferior surface of the inferior nasal turbinates was performed and CBF was determined. Beat pattern was slightly curved and rigid, not wide, and metacronic in all the observed fields analyzed. CBF was 8.2 Hz in average (reference value, 10–15 Hz) Ultrastructural assay revealed absence of the inner dynein arms in 97% of the cilia observed. The final infertility accurate diagnosis was achieved by the study of nasal CBF and ultrastructure contributing to the patient health management and genetic counseling while deciding fatherhood. Beyond this particular case, the present report may open a new field of studies in male infertility, mainly in cases of asthenozoospermia.

Bronchoalveolar Wash: Proposal of a Cutting Point for Lactate Dehydrogenase in the Differential Diagnosis of Pulmonary Diseases

Background: The analysis of bronchoalveolar lavage (BAL) is of great clinical utility in the diagnosis of pulmonary diseases (PDs). The cytological study (total and differential cell count) performed routinely in these samples, has a high orientative power and is diagnostic in some cases. The study of soluble substances has provided little information. This study aimed to determine the cut-off point of lactate dehydrogenase (LDH) activity in the diagnosis of different lung diseases. Methods: Two hundred forty-three BALs were selected from a total of 306 patients with a single respiratory disease: acute pneumonia due to common germs (NCG, n = 126), tuberculosis (TB, n = 20), mycotic pneumonia (MN, n = 12, N = 37) and interstitial diseases (IDs, n = 48). Cytological study and measurement of LDH activity were performed. The mean, standard deviation, sensitivity (S), specificity (Sp) and Youden index (YI) of this enzyme were determined. Student’s parametric and non-parametric Mann-Whitney tests were done (P < 0.05: significant). Results: Comparing the means and standard deviations of LDH in the different PDs, a significant increase of this parameter was observed in the NCG compared to the other PDs: NCG vs. TB (P = 0.003); MN, MD and ID (P < 0.0001). Based on these significant differences observed, the cut-off point for LDH in NCG was evaluated. Different values were analyzed: LDH: ≥ 150 IU/L with S: 70%, Sp: 85% and YI: 0.55; ≥ 130 IU/L with S: 77%, Sp: 80% and YI: 0.57; ≥ 120 IU/L with S: 80%, Sp: 77% and YI: 0.57 and ≥ 100 IU/L with S: 86%, Sp: 74% and YI: 0.60. Conclusions: It is proposed to perform the measurement of LDH activity for the differential diagnosis of NCG in the PD, since its mean value was significantly higher than the rest of the PD, using a cut-off point of LDH ≥ 100 IU/L; it showed a higher S and YI for the diagnosis of NCG screening. Increased LDH activity in NCG could be associated with the high number of leukocytes present in this pathology, superior to the rest of the PD. Measurement of LDH activity along with cell count would contribute to the diagnosis of PD.

Urothelial cells in smears from cervix uteri.

Objectives: To establish the cytological criteria to identify the urothelial cells in cervical smears in order to avoid mistakes in the cytological diagnosis. Materials and Methods: Cervical smears from 34 post menopausal women with vesicovaginal fistulas, advanced bladder prolapse and genital erosive lichen planes (vulvar kraurosis) (Group 1) and transitional cell metaplasia of the cervix (TCM, Group 2) were stained with Papanicolaou technique. The cervical samples were taken during the routine annual examination for prevention of the uterine cancer. Results: The smears of cervix from Group 1 showed urothelial cells from the three layers of the transitional epithelium. The umbrella cells are the bigger ones with relatively large nuclei. Frequently, they are multinucleated with single or multiple nucleoli and a typical “frothy” cytoplasm (cytoplasmic vacuoles). The cells of the Group 2 showed nuclei with oval to spindled shapes, some tapered ends, less cytoplasm than squamous metaplastic cells, powdery chromatin, small nucleoli and nuclear grooves. Conclusions: The umbrella cells may be mistaken for dysplastic cells originating in low grade squamous intraepithelial lesions lesions (LSILs) due to their nuclear and cytoplasm sizes. Therefore, it is important to know the possibility of their appearance in the cervical smears, especially in post menopausal patients in order to avoid a false diagnosis of an intraepithelial lesion. It is unlikely that deeper cells of urothelium would be confused with high grade squamous intraepithelial lesion (HSIL) cells. However, their presence might be a reason of mistake in the diagnosis. TCM is an under-recognized metaplastic phenomenon of the cervix and vagina, which is a mimicker of high-grade squamous intraepithelial lesion. The differential characteristic between umbrella cells, cells from TCM and the deeper urothelial cells, and LSIL and HSIL are detailed in the present paper.