Critical steps during the prilling process of molten lipids: Main stumbling blocks due to pharmaceutical excipient properties.

Prilling by ultrasonic jet break-up is an efficient process to produce perfectly spherical microparticles homogeneous in size. However, the material properties could affect the manufacturability and the final product properties especially with lipid-based excipients which often exhibit complex structural properties. This work presents the characterisation of six lipid-based excipients differing by their melting point and polymorphic behaviour which were used to produce microspheres using a pilot-scale prilling equipment. The experimental results were compared to theoretical calculations, especially the droplet solidification time which is a key-parameter for this process. This work highlighted that monotropic polymorphism of excipients and supercooling effect have a significant impact on process parameters which should be considered with care during formulation design.

Critical parameters involved in producing microspheres by prilling of molten lipids: from theoretical prediction of particle size to practice.

The aim of this work was to identify the key parameters which influence the running of the prilling process with lipid material from the initial melting to the formation of solid microspheres. The microsphere size would essentially result from break-up at the Rayleigh-Weber's wavelength which mostly depends on the liquid properties (mass density, surface tension and dynamic viscosity). After molten liquid extrusion through the nozzle, the cooling rate is very fast and the instantaneous temperature of the liquid jet decreases rapidly of 0.2-0.3 °C during the duration of the droplet formation (1-2 ms). This leads to no significant modification of the physical characteristics of the lipids and only a very slight change in the dynamic viscosity. Consequently, no significant effect on the optimal wavelength λ(W) and on droplet formation can occur. However, coalescence of liquid droplets has been observed during their fall, probably caused by turbulence into the air column, leading to a minor population of larger microspheres.

Pneumocystis jirovecii genotypes and granulomatous pneumocystosis.

This study describes the initial data concerning molecular typing of Pneumocystis jirovecii in a patient having developed granulomatous Pneumocystis pneumonia (PCP). Three types, B(1)a(3), B(1)a(4), B(1)b(2), were identified. All three had been described in reports concerning patients with common diffuse alveolar PCP. The present data show that identical microorganisms can be involved in both granulomatous PCP and diffuse alveolar PCP and that the pathogenesis of the granulomatous response to P. jirovecii may more likely be related to host factors.

[Granulomatous pneumocystis pneumonia]. / Pneumocystose à forme granulomateuse.

INTRODUCTION: Granulomatous pneumocystis pneumonia (PCP) is a rarity whose presentation may be misleading. CASE REPORT: We report the case of a patient suffering from chronic lymphatic leukaemia presenting with acute respiratory insufficiency, with a fatal outcome, due to granulomatous PCP. Broncho-alveolar lavage (BAL) remained negative and the diagnosis could only be made later by open lung biopsy. CONCLUSION: Although infrequent, granulomatous PCP should be recognised because in this situation the standard investigation (BAL) may be negative. New, more sensitive, methods of investigation such as the polymerase chain reaction (PCR) may permit earlier diagnosis.

Expression of TTF-1 and cytokeratins in primary and secondary epithelial lung tumours: correlation with histological type and grade.

AIMS: To assess cytokeratin (CK) and thyroid transcription factor (TTF)-1 expression in primary epithelial lung tumours by comparison with non-pulmonary carcinomas and to correlate it with their histological type and grade. METHODS AND RESULTS: Immunohistochemistry using antibodies against CKs 5/6, 7, 19, 20 and TTF-1 was applied to 165 primary and 37 secondary epithelial lung tumours. CK5/6 is a sensitive and specific marker of lung squamous carcinomas being positive in 100% of cases. CK7 is a common marker of primary lung adenocarcinomas (100% of cases) but with a lower specificity since it is also observed in other primary lung carcinomas (70% of large-cell neuroendocrine carcinomas, 40% of large-cell carcinomas, 23% of squamous carcinomas) but also in 27% of non-pulmonary adenocarcinomas. Addition of an anti-CK20 may be useful to prove or disprove the pulmonary origin of an adenocarcinoma when there is a history of colon cancer. CK19 is ubiquitous but a predominant or exclusive 'dot-like' pattern is very suggestive of high-grade neuroendocrine carcinoma. TTF-1 is a very sensitive and specific marker to document the pulmonary origin of an adenocarcinoma if a thyroid origin is excluded. Its expression in neuroendocrine lung tumours depends on the tumour grade. CONCLUSIONS: Immunohistochemical expression of CKs and TTF-1 may be correlated with histological type and grade of lung primary epithelial tumours and may allow them to be distinguished from non-pulmonary carcinomas.

[Nodular bronchioloalveolar carcinoma at early stage]. / Carcinome bronchioloalvéolaire nodulaire de stade IA.

INTRODUCTION: The aim of our study was to describe the main characteristics of bronchioloalveolar carcinoma (BAC) in the light of the latest WHO classification. The clinical, pathological and radiological features of 16 consecutive resected cases of early BAC (stage IA) were reviewed. METHODS: Retrospective pathology of 249 adenocarcinomas over a 32 months period. RESULTS: Computerised tomography (CT) features which supported a diagnosis of BAC included a peripheral location, irregular margins forming a star pattern, pleural tagging, ground-glass attenuation and an air bronchogram. Immunochemistry was positive in all cases for cytokeratins 7 and 19, EMA and TTF-1, but was only variably positive for ACE, P53 and MIB-1. For 14 out of 16 patients clinical outcome was favourable with no evidence to date of recurrence since surgery. Of the others, one developed extra-thoracic metastases and the other a local recurrence requiring further surgery. CONCLUSION: Recognising these characteristics of BAC is important as early diagnosis and treatment of this condition can be associated with an excellent prognosis.

[Congenital mesothelial cyst of the diaphragm: imaging aspects. Apropos of 2 cases in adults and review of the literature]. / Kyste congénital mésothélial du diaphragme: aspect en imagerie. A propos de 2 observations chez l'adulte et revue de la littérature.

The authors report 2 cases of congenital mesothelial cyst of the diaphragm diagnosed in adults. The differential diagnosis will be discussed and the literature reviewed.

Polymerase chain reaction on cerebrospinal fluid cells in the detection of leptomeningeal involvement by B-cell lymphoma and leukaemia: a novel strategy and its implications.

In the search of B-cell lymphoma/leukaemia dissemination to cerebrospinal fluid (CSF), we used the highly sensitive semi-nested PCR (snPCR) for the analysis of IgH gene rearrangements. This method detects a rearranged IgH gene from a single B lymphocyte which may or may not represent the neoplastic B-cell population. We therefore performed multiple snPCR (three to five) experiments on the same CSF sample, postulating that the detection of a band of the same size and sequence in different PCR runs was highly indicative of a clonal population. 17 consecutive cases with a differential diagnosis of primary (PCNSL) (n=10) or secondary (SCNSL) (n=7) CNS lymphoma or leukaemia were investigated by the new strategy. The clonal nature of the B-cell population was confirmed in 3/10 of suspected PCNSL, and in six other cases the PCR study was indicative of reactive lymphocytosis. One case revealed a clonal B-cell population in the clinical context of an autoimmune disorder. Evidence of clonal B-cell population was found in 4/7 of suspected SCNSL. In one of these cases the detected band and its sequence proved identical to that of the primary nodal lymphoma. We believe that the evaluation of B-cell clonality in CSF requires multiple snPCR amplification on the same sample to compare the size of the products and, if necessary, the DNA sequences to ascertain the diagnosis of malignancy in equivocal cytologic and clinical findings.

[GC12, marker of cells of mesodermal origin. Value and application to cytodiagnosis of serous effusions]. / GC12, marqueur des cellules d'origine mésodermique. Intérêt et application au cytodiagnostic des épanchements de séreuses.

Monoclonal antibody GC12 was examined for its value in cytopathologic diagnosis. Its sensitivity and specificity in staining reactive mesothelial cells in serous effusions was determined using the indirect immuno alkaline phosphatase anti-alkaline phosphatase technique. Smears prepared from 78 serous effusions (15 benign, 54 malignant, 9 atypical) were immunostained with GC12 and five other monoclonal antibodies: KL1 directed against cytokeratins, D33 against desmin, E29 against epithelial membrane antigen, Calam 27 against epithelial surface antigen and NEO 723 reactive with carcinoembryonic antigen. GC12 reacted positively with 80 to 100% mesothelial cells from the 15 benign effusions. Carcinoma cells were negative in 87% of malignant cases. Ovarian carcinomas were specially stained among positive cases. One malignant mesothelioma was positive and one was unreactive with GC12. Moreover, the reactivity of atypical cells in 9 serous effusions with GC12 was helpful to characterize benign and malignant effusions: the immunophenotype GC12-, Desmine-, Calam 27+, EMA+, ACE+ allowed detection of carcinoma cells in 2 cases. It is concluded that GC12 is a good tool for distinguishing carcinoma cells from reactive mesothelial cells in serous effusions.

Acquired localized elastolysis associated with varicose veins.

Localized elastolysis refers to a group of heterogeneous conditions which includes cutis laxa. We report a form of localized and non-inflammatory elastolysis evocative of cutis laxa, localized to the anteromedial aspect of the left thigh and knee, acquired at the age of 8 years and associated with unilateral adjacent venous dilatations appearing at the age of 39 years. Our case is clinically and ultrastructurally compatible with cutis laxa. Localized forms of cutis laxa not preceded by inflammation are exceptional; localization on the leg has never been described. This clinical form of elastolysis calls into question the classification of acquired elastolysis. The adjacent varicose veins also raise the question of the risk of subcutaneous damage in this condition.

[Typing of human papillomaviruses in intra-epithelial malpighian lesions of the cervix uteri: correlation with cytohistologic grade and progression of the lesions]. / Typage des papillomavirus humains (PVH) dans les lésions malpighiennes intra-épithéliales du col utérin: corrélations avec le grade cytohistologique et l'évolution des lésions.

The clinical course of condylomas of the cervix, and more generally of all cervical dysplasias whether associated with koilocytosis or not, remains unpredictable. Based on a series of earlier publications, we undertook a routine typing for human papillomaviruses in intraepithelial malpighian lesions of the cervix. We examined the real usefulness of such typing to determine prognosis of the lesions and correct clinical management. We determined the correlations between human papillomaviruses, as detected by the in situ hybridization method, with the histological grade of 125 cervical lesions (in 106 patients) and the clinical course (mean follow-up = 24 months) in 56% of them. Viruses 16/18 and/or 31/33/55, considered to be potential oncogenes, predominated, whatever the grade, in in situ hybridization positive intraepithelial malpighian lesions (57 out of 59 cases; 97%). In the follow-up cases, there was no difference in the clinical course of the lesions in terms of grade (simple condyloma mild or moderate dysplasia) based on the hybridization results (detection and typing). These findings suggest that routine viral typing cannot be proposed to determine the therapeutic attitude (abstention, destruction) for the management of intraepithelial lesions of the cervix.

[Comparative study of the expression of CEA and a myelomonocytic antigen (CD15) in serous effusions using two monoclonal antibodies NEO 723 and Leu M1]. / Etude Comparative de l'expression de l'ACE et d'un antigène myélomonocytaire (CD15) dans les épanchements des séreuses à l'aide de deux anticorps monoclonaux NEO 723 et Leu M1.

A comparative study of the reactivity of two monoclonal antibodies (MAb), NEO 723 (anti-CEA) and Leu M1 (CD15) was performed by immunocytochemistry on sixty five reactive effusions and sixty two neoplastic effusions, fifty eight due to metastases from carcinomas, two due to disseminations of sarcoma and two due to malignant mesotheliomas. The study of the expected reactivity of NEO 723 and the cross-reactivity of Leu M1 on exfoliated neoplastic cells in effusion fluids showed that the sensitivity of NEO 723 was superior to that of Leu M1 for the detection of carcinomatous metastases, as 78% reacted with NEO 723 versus 38% with Leu M1. Among the positive cases, the mean number of reactive cells was twice as high with NEO 723, while only three of the carcinomas no expressing CEA reacted with Leu M1. The study of the reactivity of benign and malignant mesothelial cells with these two antibodies also confirmed the absence of labelling of these cells. Thus, despite a good specificity for carcinoma, the combination of these two antibodies provides only a minor gain in diagnostic sensitivity (+5%) compared with the use of an anti-CEA antibody alone and a loss of sensitivity (-5%) compared with the combination of an anti-CEA and an anti-EMA antibodies. These results appear to justify the suppression of Leu M1 from the first panel of antibodies screening for carcinomatous cells in favour of a combination of anti-CEA and an anti-EMA antibodies. However, Leu M1 may be useful as a second-line test in order to define the primary tumour responsible for the effusion.

Immunophenotyping of mesothelial cells and carcinoma cells with monoclonal antibodies to cytokeratins, vimentin, CEA and EMA improves the cytodiagnosis of serous effusions.

This paper presents an immunocytochemical study performed on cytocentrifuged deposits from 109 peritoneal and pleural effusions including 20 transudates, 43 malignant metastatic effusions and 46 effusions containing atypical cells, unidentifiable as reactive mesothelial or malignant epithelial cells on the classical morphological criteria. A panel of four monoclonal antibodies (MAb) was used, including KL1 directed to cytokeratins (KER), V9 to vimentin (VIM), NEO 723 to carcinoembryonic antigen (CEA) and E29 to epithelial membrane antigen (EMA). In most transudates the reactive mesothelial cells coexpressed VIM and KER with a ring-like pattern for the latter proteins. In contrast, they were unreactive to anti-CEA and weakly and inconsistently reactive to anti-EMA. In malignant effusions, most carcinoma cells coexpressed EMA, CEA and KER with a predominant diffuse cytoplasmic pattern for the latter. Only a few malignant epithelial cells from five metastatic adenocarcinomas weakly expressed VIM. When used on the 46 effusions with unidentifiable cells, the panel of MAb allowed reactive mesothelial cells and malignant epithelial cells to be distinguished from each other in 39 of 46 cases (85%).

[Reflections on the systematic screening of cervico-vaginal smears rich in polynuclear cells]. / Réflexions à propos des frottis cervico-vaginaux systématiques de dépistage riches en polynucléaires.

The authors sometimes see cervico-vaginal smears with too many white cells. The question is: what is the meaning of these smears, and what must be the physician's attitude? The authors try to answer this question in this paper. They find 8 p. cent of genital tract infections in their observations.