Mutations in MYH9 result in the May-Hegglin anomaly, and Fechtner and Sebastian syndromes. The May-Heggllin/Fechtner Syndrome Consortium.

The autosomal dominant, giant-platelet disorders, May-Hegglin anomaly (MHA; MIM 155100), Fechtner syndrome (FTNS; MIM 153640) and Sebastian syndrome (SBS), share the triad of thrombocytopenia, large platelets and characteristic leukocyte inclusions ('Döhle-like' bodies). MHA and SBS can be differentiated by subtle ultrastructural leukocyte inclusion features, whereas FTNS is distinguished by the additional Alport-like clinical features of sensorineural deafness, cataracts and nephritis. The similarities between these platelet disorders and our recent refinement of the MHA (ref. 6) and FTNS (ref. 7) disease loci to an overlapping region of 480 kb on chromosome 22 suggested that all three disorders are allelic. Among the identified candidate genes is the gene encoding nonmuscle myosin heavy chain 9 (MYH9; refs 8-10), which is expressed in platelets and upregulated during granulocyte differentiation. We identified six MYH9 mutations (one nonsense and five missense) in seven unrelated probands from MHA, SBS and FTNS families. On the basis of molecular modelling, the two mutations affecting the myosin head were predicted to impose electrostatic and conformational changes, whereas the truncating mutation deleted the unique carboxy-terminal tailpiece. The remaining missense mutations, all affecting highly conserved coiled-coil domain positions, imparted destabilizing electrostatic and polar changes. Thus, our results suggest that mutations in MYH9 result in three megakaryocyte/platelet/leukocyte syndromes and are important in the pathogenesis of sensorineural deafness, cataracts and nephritis.

Analysis of living cells grown on different titanium surfaces by time-lapse confocal microscopy.

In this study we have combined fluorescence- and reflection-confocal laser scanning microscopy for the simultaneous visualization of living cells and surface topography beneath them. To this purpose we have designed a specific flow chamber and we have tested it with osteoblasts grown on an opaque, thick support, made of smooth or sandblasted titanium. Cells were loaded with Calcein-AM or tetramethylrhodamine methyl ester (TMRM), two probes employed as indicators of cell viability/morphology and mitochondrial membrane potential, respectively. Besides the acquisition of stacks of confocal sections, the system allowed also vertical views and faithful three-dimensional reconstruction of the samples. Confocal microscope implemented with our flow chamber proved to be a promising tool for time-lapse investigation of cell-biomaterial interactions.

Parasuicide in Rovigo (North of Italy) during the period 2000-2005.

INTRODUCTION: The greatest predictor of eventual suicide is parasuicide, which includes both suicide attempts and deliberate self-harm with no intent to die. The rate of parasuicide is reckoned to be at least ten times the suicide rate. METHODS: An observational study of the population of Rovigo Public Health Unit has been carried out to investigate parasuicide cases that presented to a general hospital in the six-year period from 1st January 2000 to 31st December 2005. RESULTS: An incidence of 36.39 parasuicides/100,000/year referred to a general hospital has been estimated, with a majority of female and young subjects. The principal method used was drug poisoning (59.1%); the more frequent diagnoses are mood and personality disorders. Method of attempt distribution is different for age and gender (p < 0.001), while diagnosis distribution is different only for age (p < 0.001). Comparison between method of attempt and diagnosis distribution indicates a significant difference (p < 0.01). In 36.4% of cases there was no contact with the Mental Health Service after parasuicide. DISCUSSION: The present study confirms that parasuicide is more common in females and younger people and that the more probable diagnoses are mood and personality disorders. The finding of a high number of subjects without any previous contact with the Mental Health Service and, especially, after parasuicide, claims attention on primary and secondary prevention of suicidal behaviour. CONCLUSIONS: The results appear to be in line with those from literature on parasuicide in Western populations.

Thrombocytopenia, giant platelets, and leukocyte inclusion bodies (May-Hegglin anomaly): clinical and laboratory findings.

PURPOSE: May-Hegglin anomaly is a rare hereditary condition characterized by the triad of thrombocytopenia, giant platelets, and inclusion bodies in leukocytes. Clinical features and the pathogenesis of bleeding in this disease are poorly defined. PATIENTS AND METHODS: From 1988 to 1996 we studied 15 new May-Hegglin anomaly patients from 7 unrelated Italian families. In addition to clinical examination and routine laboratory testing, we measured bleeding time, platelet aggregation and release reaction, and platelet staining for tubulin, and performed ultrastructural study of polymorphonuclear leukocytes. RESULTS: Although the mean age of our patients was 33 years, May-Hegglin anomaly had not been previously recognized in any of them. Bleeding diatheses ranged from severe to absent, and platelet count from 26 to 178 x 10(9)/L. No correlation was found between bleeding tendency and platelet count. Previous therapy with corticosteroids, high-dose immunoglobulins, and splenectomy had no effect on platelet count or bleeding diathesis. Desmopressin infusion greatly shortened the bleeding time in the most severely affected patient. The in vitro function of platelets was normal except for the absence of shape change in all subjects and defective response to epinephrine in 8 of 15 patients. Platelet tubulin was distributed unevenly instead of being organized in a circumferential band at the cell periphery. CONCLUSION: The diagnosis of May-Hegglin is easily missed, and its frequency is probably underestimated. A qualitative defect of platelets may be responsible for mild bleeding diathesis even in the absence of thrombocytopenia, while severe bleeding results from both qualitative and quantitative platelet defects. May-Hegglin anomaly should be suspected whenever a patient has a low platelet count or a bleeding diathesis of unknown origin.

Comparison of annexin V and calcein-AM as early vital markers of apoptosis in adherent cells by confocal laser microscopy.

Although morphological criteria for apoptosis are in general reliable, no systematic comparison of the techniques employed thus far has yet been performed. In this study, using confocal laser microscopy, we compared the performance of annexin V-FITC and calcein-AM for early detection of apoptosis in living adherent cells. Experiments were carried out on two distinct cell lines, PC 12 and NIH3T3, endowed with different shape and adhesion properties. The apoptotic process was followed for a prolonged period in the same cells of a predetermined field by means of a special flow chamber. Our results show that both probes allowed the detection of apoptotic cells in either cell line. However, some cells that clearly exhibited apoptotic changes on calcein visualization were annexin-negative. In NIH3T3 cells, annexin negativity of apoptotic cells was correlated with the preservation of cell shape and adhesion properties. These findings show that, at least in PC12 and NIH3T3 cells, annexin might be less sensitive than calcein-AM for early apoptosis detection and, for NIH3T3 cells, suggest that phosphatidilserine exposure is in some way linked to changes in cell shape and/or adhesion to culture substrate. (J Histochem Cytochem 46:895-900, 1998)

Beta-TG and plasma catecholamines levels after sympathetic stimuli in hypertensives and patients with peripheral vascular disease.

The influence of the sympathetic nervous system on platelet functions in vivo is still controversial. The aims of our study were to compare the response to various sympathetic stimuli in normal subjects and in patients with essential hypertension (HT) or peripheral vascular disease (PVD) and to evaluate any correlations among plasma levels of catecholamines, beta-thromboglobulin (beta-TG) and platelet factor 4 (PF4). In basal conditions beta-TG and PF4 values in the HT patients were higher than those observed in the controls of the same age but lower than those of the PVD patients. Although the different sympathetic stimuli (90 degrees tilting, handgrip, treadmill test, bicycle test) caused a significant increase of the plasma epinephrine (E) and norepinephrine (NE) levels, they did not modify the beta-TG and PF4 levels in any of the groups studied. The platelet activation indices, therefore, regardless of the basal values, do not seem to be influenced by sympathetic stimulation.

A comparison of accuracy of computer treatment planning systems in brachytherapy.

An intercomparison was made between dose-rate point calculations with five commercial computer treatment planning systems used in brachytherapy. Sixteen Italian radiotherapy medical physics institutions performed a twofold test, respectively for point and linear sources, reproducing actual clinical implants and the results were compared to references values. The test was designed in order to determine the errors introduced separately by computation and by implant reconstruction via orthogonal films. The results show that for point sources a better accuracy can be achieved than for linear sources. It is also shown that a large variation of results exists between computation systems and the variability is larger for linear sources. The digitizing procedure adds a large amount of error. In the whole set of calculation points, the overall percent difference between computed and reference dose-rate values is larger than +/- 5% in 18% cases for point and in 52% cases for linear sources, with data input by digitizer. It seems that relevant errors do occur when computing actual dose rates, resulting in clinically relevant inaccuracies in the calculated absorbed dose and in its relative effectiveness.

Quality assurance in radiotherapy: the importance of medical physics staffing levels. Recommendations from an ESTRO/EFOMP joint task group.

The safe application of ionising radiation for diagnosis and therapy requires a high level of knowledge of the underlying processes and of quality assurance. Sophisticated modern equipment can be used effectively for complicated diagnostic and therapeutic techniques only with adequate physics support. In the light of recent analyses and recommendations by national and international societies a joint working group of representatives from ESTRO (European Society for Therapeutic Radiology and Oncology) and from EFOMP (European Federation of Organisations for Medical Physics) was set up to assess the necessary staffing levels for physics support to radiotherapy. The method used to assess the staffing levels, the resulting recommendations and examples of their practical application are described.

Platelet activation indices and apolipoproteins in hypertensive patients.

We have studied the platelet activation indices beta-thromboglobulin (beta-TG and platelet factor 4(PF4), triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), low-density lipoprotein (LDL), very-low-density lipoprotein (VLDL) and apolipoprotein (A1, A2, B, C2, C3, E) profiles of 22 untreated essential hypertensive subjects (WHO stages 1 and 2) and 22 controls, to see if there might be some causal relationship between lipoprotein abnormalities and greater platelet activation. The results showed the patients had both greater platelet activation than the controls, as demonstrated by higher plasma beta-TG levels (P less than 0.01) and lower apolipoprotein A2 levels (P less than 0.05). However there were no significant correlations between the platelet activation indices and the plasma levels of apolipoproteins, lipoproteins or lipids in either group.

An in vitro model for the assessment of manganese neurotoxicity.

PC12 (undifferentiated and differentiated) and C6 cells have been used to investigate kinetics, morphological and functional endpoints following exposure to MnCl(2) and manganic transferrin (Mn-Tf). [Mn](i) in undifferentiated (non-differentiated cells) exposed to both free (MnCl(2)) and bound Mn (Mn-Tf), was three- to fivefold lower as compared to differentiated (differentiated) PC12 cells and higher by one order of magnitude as compared to glial C6 cells. Exposure to both MnCl(2) and Mn-Tf was followed by time- and dose-dependent morphological changes characteristic of apoptosis, which was never observed in Mn-exposed C6 glial cells. Results from cell viability assays were consistent with apoptotic response rates quantified by cell count. Threshold concentrations for undifferentiated and differentiated PC12 cells were 10(-6) and 10(-5)m, respectively. Thus, despite their greater ability to accumulate Mn, differentiated PC12 cells are less sensitive to Mn-induced apoptosis. This model might be relevant to neuronal degeneration induced by Mn occurring in the developing brain and possibly in clinical manganism. Such critical doses at the cellular level seem to be consistent with Mn levels (5x10(-6)m) recorded in the basal ganglia of monkeys chronically exposed to Mn and developing clinical signs of manganism.

Prognostic value of hyponatremia in patients with severe chronic heart failure.

In order to evaluate the incidence and the prognostic value of hyponatremia (hypoNa) in patients (pts) with severe chronic heart failure (SCHF), the authors studied 161 consecutive pts (113M, 48F ages sixty-seven +/- ten) with SCHF in NYHA class III-IV. The cause of SCHF was ischemic in 64 pts, hypertensive in 39, valvular in 14, alcohol-related in 3, and idiopathic in 41. Pretreatment hypoNa (less than 135 mmol/L) was found in 64/161 pts (40%) (Group I); Na+ was less than 125 in 10 pts, 125-130 in 19, and 131-135 mmol/L in 35; 42/64 pts (66%) of Group I were in NYHA class IV at admission. In the pts with pretreatment Na+ less than 125 mmol/L, hypoNa was persistent and refractory to high-dose furosemide (less than 500 mg/day) and water restriction. Cardiovascular mortality of Group I pts was 69% within twenty-four months (34 pts died of low-output syndrom and 10 suddenly). All pts with Na+ less than 130 mmol/L died within six months. The 20 pts who normalized Na+ are alive, and in NYHA class II-III (follow-up: twenty-six +/- fifteen, six to sixty months). Pts without hypoNa were 97/161 (Group II), and 58/97 (60%) are alive (follow-up: thirty +/- eighteen, five to fifty-eight months), whereas 39 pts died (27 suddenly, 9 of low-output syndrome, and 3 of extracardiac disease) within twenty-four months. The mortality rate of Group II was significantly lower (40% vs 69%, p less than 0.001) compared with Group I. The two groups were similar for age, sex, and cause and duration of SCHF.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of high-dose furosemide in refractory congestive heart failure.

High-dose firosemide is considered effective in primary renal sodium retention but is not generally recommended in congestive heart failure. In order to evaluate efficacy and safety of high-dose furosemide (greater than 500 mg/day), the authors studied 20 patients (pts) resistant to therapy (including furosemide less than 500 mg/day) selected from 161 pts admitted for chronic heart failure. All refractory pts (15 men and 5 women, mean age sixty +/- 12 years) were in NYHA class IV and showed hyponatremia (130 +/- 5 mEq/L) and impaired renal function (BUN 31 +/- 14 mg/dL, serum creatinine 1.3 +/- 0.3 mg/dL and BUN/creatinine ratio 23 +/- 7). In addition to digitalis, dopamine, angiotensin-converting enzyme inhibitors, or vasodilators, IV high-dose furosemide (775 +/- 419 mg/day, 500-2000) was given for ten +/- five days under daily clinical and laboratory monitoring. Three pts died of low-output syndrome while 16 pts were upgraded to NYHA class III and 1 pt to class II; a mean weight reduction of 7.3 +/- 2.9 kg in ten + five days (0.80 +/- 0.4 kg/day) and a mean diuresis increase of 88 +/- 57% occurred. The maximal dose of furosemide did not correlate with serum creatinine but did correlate with BUN/creatinine ratio (r = 0.78, p less than .001). Pts were discharged on with chronic heart failure, and 43% in the subgroup in NYHA class IV with hyponatremia. High dose furosemide was effective for rapid removal of excess water and salt in "furosemide-resistant" congestive heart failure. The relationship between renal impairment and maximal furosemide doses seems to confirm the role of renal pharmacokinetics in the appearance of furosemide resistance.

Measurements of radiocesium transfer to milk and calculation of resulting dose in Brescia, Italy, following the Chernobyl accident.

Results are presented of several measurements on components of the cows' milk chain performed at our Medical Physics Service after the Chernobyl accident. Values were obtained for Cs isotope transfer coefficients, namely, for cows' diet-milk and diet-feces transfers. Other measured parameters were the effective half-life of Cs in milk and the 134Cs:137Cs ratio. In addition, an evaluation of Cs contribution to the absorbed dose to population from milk is performed.

[The autonomic nervous system in myocardial infarction]. / Il sistema nervoso autonomo nell'infarto miocardico.

The occurrence of alterations in the autonomic control of cardiovascular function has long been known. Nevertheless, only in the last 15 years with the development and utilization of techniques such as heart rate variability and baroreflex analysis, unforeseeable amounts of information have been collected. The appraisal of alterations in the autonomic control mechanisms in the acute and post-acute phase of myocardial infarction has not only confirmed the presence of an increased sympathetic and of a reduced vagal modulation in most of post-myocardial infarction patients. It has also allowed the clinically relevant identification of those patients with an increased arrhythmic and total cardiac mortality. Most the high-risk patients are indeed characterized by a marked reduction in heart rate variability that can be detected using different methodologies. For example, with time-domain analysis of heart rate variability, it has been shown that an SDNN < 70 ms identifies patients at risk. By using spectral analysis, it has been possible to realize that the loss or marked attenuation of those rhythmical components that reflect autonomic modulation is also a characteristic of the high-risk patients. A decreased responsiveness of the sinus node to autonomic inputs may also explain the reduction in baroreflex sensitivity that characterizes patients with an increased arrhythmic mortality. In conclusion, the appraisal in each patient of the extent of the alterations in the autonomic control mechanism is nowadays a critical component of the clinical assessment not only for risk stratification but also in order to guide the prescription of new pharmacological and non-pharmacological therapies.

[The approach to the infradiaphragmatic and adrenal aorta]. / Vie di accesso all'aorta pararenale e sottodiaframmatica.

The progressive selection of the patient reserved to open surgery is determined by the preference of the endovascular option on the treatment of abdominal aortic aneurysm. This fact probably will cause a reduction of the number of cases singularly approachable by the infrarenal way while will prevail the need of control and clamping of the infradiaphragmatic and adrenal aorta. The approach to the infradiaphragmatic and adrenal aorta introduces technical and physiopathologic problems related to the hold interconnection between the vessel and the surrounding structures, particularly those visceral. The choice of the approach is determined by the surgical gesture that is had to perform. The simple supraceliac aortic clamping or the wrapping of bypass on the visceral arteries can be made by an anterior approach, supramesocolic transperitoneal, while the correction of an juxtarenal or adrenal aneurysm can be performed by a submesocolic approach. The extraperitoneal approach offers a complete vision of this part of aorta, without the pancreas or left renal veins interposition, but it allows a bad control of the iliac and right renal arteries. The surgeon has to have familiarity with both the approaches, to be able to perform his own intervention in the simplest and effective way.

Heart rate and systolic blood pressure in patients with persistent atrial fibrillation. A linguistic analysis.

BACKGROUND: During atrial fibrillation (AF), ventricular response is highly irregular and thus the beat-to-beat variation of blood pressure is increased because of variations in filling time and in contractility. OBJECTIVES: Aim of the present study is to investigate the short-term dynamics of RR and SAP series in patients with AF, during rest and tilt, and after restoration of sinus rhythm. METHODS: We computed symbolic sequences of the three phases, as they retain important features of the dynamics generated by the underlying control system. Then we applied a method based on rank order statistics of symbolic sequences to investigate the profile of different types of dynamics. The linguistic distance (range 0-1) between sequences represents a measure of similarity to assess whether the different physiological states are reflected on the dynamics of RR and SAP series. RESULTS: The distance between rest and tilt phases is 0.06 ± 0.02 for RR series, meaning they are very similar, while it is 0.21 ± 0.13 for SAP series, showing a difference in the short-term dynamics. RR mean decreases during tilt (738 ± 164 vs. 692 ± 152 ms, p <0.05, rest vs. tilt), while mean SAP is not significantly different (101 ± 20 vs. 104 ± 14 mmHg, rest vs. tilt). Comparing AF and sinus rhythm, both RR and SAP series result different in terms of the computed distance. CONCLUSIONS: SAP short-term dynamics seem to significantly change when comparing rest and tilt phases, while RR series remain unchanged. Moreover, RR mean but not SAP series significantly decreases during tilt.