Body weight variation is not an independent factor in the determination of functional hypothalamic amenorrhea in anorexia nervosa.

OBJECTIVE: Functional hypothalamic amenorrhea (FHA) is one of the foremost manifestations in anorexia nervosa (AN), but a subset of patients have menses despite marked weight loss and underweight. The aim of our study was to investigate parameters potentially influencing FHA in AN. DESIGN AND METHODS: In this observational retrospective study, we selected 114 female patients with AN who completed a 12 months semi-residential rehabilitation program and a subsequent 12 months outpatient follow-up. We divided our sample into three groups: "Group 0" patients who experienced FHA and recovered their menses, "Group 1" persistent FHA, "Group 2" never experienced FHA, and looked for clinical and hormonal correlations. RESULTS: At the enrollment, the BMI was higher in Group 2 than in Group 1 (p = 0.0202), but the last follow-up weight was higher in Group 1 (p < 0.0001) despite persistent amenorrhea. At logistic regression, the higher BMI at which patients experienced amenorrhea was the main prediction factor for persistent FHA. Notwithstanding comparable leptin levels at admission, they improved significantly at discharge only in Groups 0 and 2 (p = 0.0054 and p = 0.0104, respectively). FT3 at admission was significantly higher in Group 2 than in Group 0 (p = 0.0249). CONCLUSIONS: FHA does not correlate strictly with body weight variations in AN patients, indicating a multifactorial origin, likely including an individual predisposition. Higher FT3 levels identify patients who continue having menses at extremely low BMI. AN patients with persistent FHA constitute a subgroup in whom estroprogestins should be considered after significant weight recovery to prevent prolonged tissue hypoestrogenism.

Mixed-lineage leukemias and phenotypic shifts occurring in relapsed cases of acute T lymphoblastic lymphomas.

Specimens from 19 patients with NHL were also phenotyped at the onset of the disease; among them, 9 were studied in the relapse phase. The analysis was carried out with monoclonal antibodies directed against T and myeloid cells; at diagnosis, all cases presented an immature thymic phenotype. When analyzed at relapse, phenotypic changes were observed: intra-lineage dedifferentiations (6 cases); mixed-lineage lymphoid and myeloid (2 cases), and pure myeloid relapses (1 case). The molecular analysis of the TCR-genes configuration showed a germ-line pattern at onset and relapse in Case 9 and a modification of the rearrangement patterns during the evolution of the disease in Case 6. These data point out that the relapse is often accompanied by intra-lineage modifications resembling dedifferentiation and, more rarely by a myeloid switch. The phenotypic follow-up of these patients may be important to the implementation of chemotherapeutic protocols that are more adequate for the biological evolution of the disease.

Peripheral neuropathy in macroglobulinemia: incidence and antigen-specificity of M proteins.

Peripheral neuropathy was found in 12 (46%) of 26 patients with macroglobulinemia. The neuropathy was subclinical in two. Anti-myelin-associated glycoprotein (MAG) activity was found in six (50%) patients with neuropathy. Sural nerve biopsies showed demyelination and IgM deposits on the myelin sheath. In one patient who had no anti-MAG activity, the serum IgM bound to peripheral myelin by indirect immunofluorescence and to several protein bands in peripheral nerve and other tissues by immunoblot. In the other five patients with neuropathy, we found no binding of M proteins to nerve components, but in three patients there were endoneurial IgM deposits in nerve biopsy. Peripheral neuropathy may be related to the antigen-specificity of M proteins.

Monocytic differentiation induced by 1,25 dihydroxyvitamin D3 in myeloid cells: an ultrastructural immunocytochemical study.

We have studied, by ultrastructural morphology and immunocytochemistry, the alterations that occur in cells from the HL60 leukaemia cell line and from patients with CGL following incubation in vitro with 1,25(OH)2D3 for 2-5 days. The main morphological changes observed were in the nuclear shape, the development of autophagic vacuoles and the appearance of a population of small granules in the cytoplasm. These changes were associated with a significant reduction in MPO activity and increased expression of membrane antigens detected by the monocyte-specific McAb FMC17 and FMC32, as shown by the IGM at EM level, and a decrease in granulocyte-specific antigens demonstrated by the McAb FMC10. These observations suggest that promyelocytes and myelocytes could transform into monocyte-like cells and that this remodelling of cells was associated with autophagic digestion of cellular structures.

Characterization by ultrastructural cytochemistry of normal and leukemic myeloid cells reacting with monoclonal antibodies.

Six monoclonal antibodies (McAb) of the FMC series with specificity for granulocytic or monocytic cells were investigated by means of the immunogold method in combination with the myeloperoxidase reaction at ultrastructural level. This method allowed a precise characterization of the level of myeloid maturation at which the antigens detected by the McAb were expressed and confirmed their specificity for the two myeloid lineages. In addition, it facilitated the identification of granulocytic and monocytic cell components in cases of acute myeloid and chronic myelomonocytic leukemia. Blast cells did not react with any of the McAb used, and the reactivity in more mature cells was parallel to the development of cytoplasmic granules. The pattern of reactivity was similar in normal and leukemic cells, thus suggesting that these McAb could be applied to studies of normal myelopoiesis and to the classification of leukemia.

Biphenotypic leukaemia: a case of mixed T lymphoblastic and myeloblastic leukaemia.

A case of mixed acute leukaemia with T lymphoblastic, myeloblastic, and monocytic components is described. The use of immunological markers, ultrastructural morphology, cytochemistry, immunochemistry, and combined techniques, simultaneously detecting two markers in individual cells, made it possible to define the different blast cell populations.

The radiation dose to the lens in radiology of the orbit.

This paper describes research into measurement and reduction of the radiation dose to the lens during various examinations, namely skull and orbit, optic canal and optic strut, superior and inferior orbital fissure, localisation of foreign bodies in the eye, calcifications, orbital fractures, macrodacryography and orbital venography. Using rare-earth screens and high-sensitivity films, without an antiscattering grid, and with an added filtration of 0.5 mm Cu, it is possible to reduce the radiological risk during all investigations involving skull, orbit and eyeballs, while maintaining a good image quality. Particularly in those examinations with direct magnification (macrodacryography and venography, foreign bodies in the eye, orbital fractures), the dose to the lens is very low: less than 0.2 mGy/radiograph.

Light and electron microscopic study of cyclosporin A-induced gingival hyperplasia.

Four transplant (3 kidney, 1 bone marrow) patients with cyclosporin A (CyA)-induced gingival hyperplasia are described. Light and electron microscopic findings of gingival biopsies showed in all patients that, in addition to an increase of collagen, CyA induced in the subepithelial space an enormous infiltration of morphologically normal plasma cells in different stages of maturation. These data, together with the reversibility of the lesion upon discontinuation of the drug, suggest that individual hypersensitivity is probably the most acceptable explanation of CyA-induced gingival hyperplasia. This hypothesis is also discussed in relation to the CyA-suppression of the functions of some T-lymphocyte subsets.

Ultrastructural alterations of megakaryocytes in some hematologic disorders.

Bone marrow and peripheral blood samples of patients affected by hematologic disorders involving the megakaryocytic line were examined at the electron microscope. While in idiopathic thrombocytopenic purpura the megakaryocytes appeared almost normal, in primary thrombocythemia there were various ultrastructural alterations interpreted as a counterpart of ineffective thrombocytopoiesis. In one patient with blastic chronic myeloid leukemia and in another with acute megakaryoblastic leukemia, the electron microscope study was very useful in the identification of immature megakaryocytes. In fact, analysis of various ultrastructural parameters allowed some blast cells to be attributed to the megakaryocytic series. Furthermore, platelet demarcation system abnormalities and alterations of the circulating thrombocytes confirm the hypothesis that there is a block in megakaryocytic maturation.

Morphologic, immunologic, and cytogenetic characteristics of secondary acute unclassifiable leukemia in Hodgkin's disease.

Blast cells from five cases of secondary unclassifiable leukemia following therapy for Hodgkin's disease were studied by cytochemical, immunological and cytogenetic analyses. Cytochemical and immunological reactivity were in accordance with poorly differentiated, myeloid blasts. The four cases in which karyotype analysis was performed showed specific chromosomal abnormalities. No evidence of multiple lineage involvement was found. Problems in classifying these cases of secondary ANLL were due to the high grade of undifferentiation of the blast cells. Their low cytochemical reactivity with markers of myeloid differentiation was similar to what may be observed in patients with acute undifferentiated leukemia or with chronic myeloid leukemia in blast crisis.

Usefulness of x-ray in the detection of complications and side effects after laparoscopic sleeve gastrectomy.

BACKGROUND: Laparoscopic sleeve gastrectomy (LSG) is a surgical procedure which reduces the gastric volume causing a feeling of early fullness while decreasing hunger due to a reduced secretion of ghrelin. This leads to a considerable loss of body weight. The purpose of this study was to assess the usefulness of early x-ray examination and subsequent x-ray follow-up in the detection of postoperative complications and long-term functional recovery. METHODS: From March 2010 to April 2011, 101 consecutive patients underwent LSG for morbid obesity and were subsequently included in this retrospective study. All patients were submitted to early x-ray examination 1-3 days after surgery and x-ray follow-up 3-6 months after surgery to detect the presence and persistence of surgical and functional complications. RESULTS: Early postoperative x-ray examination detected one case of suture leakage 1 day after surgery as well as one abscess and one gastric fistula in two patients who had become symptomatic 9 and 10 days after surgery. CONCLUSIONS: Early x-ray examination showed that complications were mainly functional and rarely surgical. Subsequent follow-up showed that functional disturbances were significantly reduced over time. X-ray is an easy and reliable method for detecting complications and side effects of LSG. However, in view of the extremely low incidence of surgical complications revealed at the early x-ray examination, this procedure may not be required as a routine examination in all operated patients and should be performed only in patients who become symptomatic and those considered at risk of developing more serious complications.

[Causes and incidence of short-life keratitis caused by dacryography. Optimization of protective parameters]. / Cause e incidenze delle cheratopatie fugaci da dacriografia. Ottimizzazione dei parametri protettivi.

This study was aimed at investigating the changes occurring in superficial corneal epithelium (localized or diffuse, dotted or linear) as observed in 300 eyes after macrodacryography with iodate contrast media (iodized oil and water soluble non-ionic agents). In our opinion, the causes of iatrogenic short-life keratitis are: needle injury, the deposition of iodate contrast medium on the cornea and the reduction of palpebral winking, favoring dry eye, due to superficial anesthesia. The pharmacologic protection of the cornea by means of high-viscosity drugs allows both the number and the degree of keratitis to be markedly reduced. Therefore, contrast media must be chosen on the basis of anamnestic and clinical data, as well as of patient's symptoms, focusing mainly on the characteristics of the various agents--i.e., density, concentration, viscosity.

A case of Waldenström macroglobulinemia terminating in acute lymphoblastic leukemia.

A case of acute lymphoblastic leukemia occurring in a patient with Waldenström macroglobulinemia more than 6 years after the onset of the disease is reported. At the time of acute transformation, bone marrow and peripheral blood lymphoid populations were almost entirely represented by lymphoblasts; the serum monoclonal peak had disappeared and no cells bearing surface or intracytoplasmic immunoglobulins were found. This observation suggests that the blast crisis might be derived from a dedifferentiative process within the lymphatic clone of the chronic phase.

Alkaline phosphatase activity in phagocytizing granulocytes: a backscattered electron imaging study.

The scanning electron microscope (SEM) in the backscattered electron imaging (BEI) mode has been used to detect alkaline phosphatase activity in normal human PMNs using a lead method. Unfortunately commercial scanning electron microscopes do not allow resolution of the small and dispersed lead depositions in the secondary granules. However, polymorphonuclear leukocytes (PMNs) previously incubated with latex particles, show well evident lead precipitates allowing identification of different phases of the phagocytic sequence. In fact the finding of irregular depositions arranged around a central not-emitting zone might be due to the secondary granules gathering near the phagocytic vacuole. Moreover, homogeneous, round, well defined precipitates, having a diameter slightly larger than latex particles, probably corresponding to vacuoles filled with the enzyme after granules coaslescence and fusion.

[Role of sophisticated radiological technics in the diagnosis of lacrimal fistulas]. / Il ruolo delle techniche radiologiche sofisticate nella diagnostica delle fistole lacrimali.

Lacrimal fistulae are rare and not well known by radiologists. Four cases are illustrated, two congenital (a skin-canaliculous and a canaliculous-palpebral fistula) and two acquired (a rare case of sac-orbital fistula and a canaliculous-palpebral fistula arisen during dacryography). Fistulae are to be distinguished from pedunculated lacrimal diverticulous of which an interesting and rare case is presented. After a careful examination of the pathogenesis and symptomatology of lacrimal fistulae the authors agree that only macrodacryography carried out by means of sophisticated techniques (seriography, magnification, subtraction) enables a precise demonstration of the originating point, of the course and outlet (cutaneous and internal) and provides therefore indispensable data for surgery.

Morphological heterogeneity of Leu7, Leu11 and OKM1 positive lymphocyte subsets: an ultrastructural study with the immunogold method.

The morphological features of normal peripheral blood lymphocytes reactive with three monoclonal antibodies (MoAb) against natural killer (NK) cells, Leu7, OKM1 (CD11b) and Leu11 (CD16) and with two anti-T cell MoAb, CD4 and CD8, have been analysed at ultrastructural level by an indirect immunogold method. Cells having the features of large granular lymphocytes (LGL) but also lymphocytes displaying different morphological characteristics (non LGL; e.g. high nucleo-cytoplasmic ratio and few cytoplasmic organelles) were seen reactive with each of the MoAb investigated. Leu7 identified a higher proportion of LGL (60-80%) than OKM1 (10-95%) and Leu11 (20-48%), and with a stronger binding. A distinct granular structure, recognized as parallel tubular arrays, was more characteristic of the Leu7+, CD8+ LGL and was less frequently seen in the OKM1 and Leu11 positive LGL subpopulation in four out of the five donors investigated. It is of interest that the Leu11 and OKM1 positive subsets, which correspond functionally to cells with greater NK function, had relatively less LGL than the Leu7 positive subsets, raising the issue of the true morphology of NK cells in man. The existence of a minority of CD4 positive LGL was confirmed. Our findings demonstrate that there is a degree of morphological heterogeneity within the normal NK lymphoid population as defined by the membrane phenotype and that certain variability among normal individuals regarding the proportion and structural features of the NK subpopulations may be present.

[Does a iatrogenic damage caused by dacryography exist?]. / Esistono danni iatrogeni da dacriografia?

This study was aimed at investigating the changes in corneal epithelium caused by dacryocystography. Such changes can be either chemical--from iodate contrast media (iodized oil and water-soluble non-ionic cm)--or physical--from needle or catheter. Twenty-five patients (50 eyes) were studied using BUT, Schirmer tests, biomicroscopy, and pachymetry. Follow-ups immediately after dacryocystography demonstrated superficial keratitis, which was less severe with water-soluble non-ionic cm. Such lesions were hardly ever observed in patients who had been treated with dacriosol ointment on the cornea just before the X-ray exam. Follow-ups at 24 hours showed the superficial keratitis to have disappeared in all patients, independent of the kind of cm employed and of the ophthalmic ointment applied to the cornea.

Immunohistochemical evaluation of bone marrow involvement in hairy cell leukemia during interferon therapy.

Bone marrow biopsies from 7 patients afflicted with hairy cell leukemia were studied with a panel of monoclonal antibodies at different intervals during alpha recombinant interferon therapy. Monoclonal antibodies Dako-LC and F 8.11.13 were used because they are highly reactive with hairy cells, and 82H3 and LeuM 1 were also utilized to identify respectively the residual hemopoietic and myeloid tissue. All antibodies are reactive on sections of formalin-fixed, paraffin-embedded biopsy material. Before therapy a uniform hairy cell infiltrate was present and very little tissue was visible even after immunohistochemical staining. During therapy, identification of hairy cells in routine biopsies was difficult, since they were mixed with normal cells and the bone marrow was generally hypoplastic. Immunohistochemical stains seem to allow better identification of hairy cells and a more precise estimate of the degree of repopulation by normal bone marrow cells. It was evident, especially after immunocytochemical analysis, that interferon drastically reduced the extent of the infiltrate and allowed recovery of normal hemopoiesis, but did not produce complete remissions.

Simultaneous demonstration of membrane antigens and cytochemistry at ultrastructural level: a study with the immunogold method, acid phosphatase and myeloperoxidase.

We describe a method for electron microscopy that combines myeloperoxidase or acid phosphatase cytochemistry with the labelling of cell surface antigens with monoclonal antibodies and colloidal gold. This technique was tested in samples of normal mononuclear cells, leukaemic T cells with a mature or immature phenotype and an acute myeloid leukaemia. This method allows the demonstration at a single cell level of ultrastructural morphology, cytochemical reaction and the presence of a membrane antigen. It will improve further the analytic power of electron microscopy in the characterization of leukaemic cells particularly in cases of mixed leukaemias and in the study of normal haemopoietic differentiation with monoclonal antibodies.

Distinct morphophenotypic features of chronic B-cell leukaemias identified with CD1c and CD23 antibodies.

Morphological criteria usually applied to diagnose various subtypes of B-cell chronic lymphoid leukaemia are largely subjective. Immunophenotyping of 61 relevant cases using a selected panel of monoclonal antibodies (mAb), showed that CD1c and CD23 mAb were able to separate B-cell chronic lymphocytic leukaemia (B-CLL) from other chronic B-cell lymphoproliferative diseases. Lymphocytes of B-CLL were CD1c-, CD23+, whereas those of other types of chronic B-cell leukaemia were CD1c+/-, CD23-, and CD38/-. Non-B-CLL cases had a significantly higher amount of large peroxidase-negative (unstained) cells analyzed with an automated blood cell counter (Technicon H6000). This type of volumetric assessment allowed a separation between typical and "atypical" B-CLL, which otherwise were both CD1c-, and CD23+. These combinations of phenotypic markers corresponded to well-defined haematopathologic entities, conventionally diagnosed on peripheral blood (PB) and bone marrow smears, and on histologic sections of lymph nodes and spleen.