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Today it is widely accepted that molecular mechanisms triggering cancer initiate with a genetic modification. However, a genetic alteration providing the aberrant clone with a growing advantage over neighboring cells is not sufficient to develop cancer. Currently, tumors are considered a heterogeneous population of cells and an extracellular matrix (ECM) that make up a characteristic microenvironment. Interactions between tumor cells and cancer microenvironment define cancer progression and therapeutic response. To investigate and clarify the role of ECM in the regulation of cancer cell behavior and response to therapy, the decellularization of ECM, a widely used technique in tissue engineering, has been recently employed to develop 3D culture model of disease. In this review, we briefly explore the different components of healthy and pathological ECM and the methods to obtain and characterize the ECM from native bioptic tissue. Finally, we highlight the most relevant applications of ECM in translational cancer research strategies: decellularized ECM, ECM-hydrogel and 3D bioprinting.

Objective Healthcare agencies recommend limited use of aerosol‐generating procedures to mitigate disease (COVID‐19) transmission. However, total dispersion patterns of aerosols, particularly respirable droplets, via dental ultrasonic units is unclear. The purpose of this study was to characterize and map total spatter, droplet and aerosol dispersion during ultrasonic scaling in simulated and clinical contexts. Methods Ultrasonic scaling was performed on dental simulation units using methylene blue dye‐stained water. All resultant stain profiles were photoanalysed to calculate droplet size and travel distance/direction. Airborne particle concentrations were also documented 0–1.2 m (0–4ft.) and 1.2–2.4 m (4–8ft.) from patients during in vivo ultrasonic scaling with a saliva ejector. Results Stain profiles showed droplets between 25 and 50µm in diameter were most common, with smaller droplets closer to the mouth. In‐vivo particle concentrations were uniformly low. The smallest (10 µm, PM10+) particles were most common, especially within 1.2 m (4ft.) of the patient. Respirable particles (PM2.5) were uncommon. Conclusions Tests showed the highest concentration of small droplets in zones nearest the patient. While uncommon, particles were detected up to 2.4 m (8ft.) away. Furthermore, observed particle sizes were consistent with those that can carry infectious agents. Efforts to mitigate the spread of inhalable aerosols should emphasize proximate regions nearest the procedure, including personal protective equipment and the use of evacuation devices.

Interest in global health is increasing among neurology residents. However, funding, time, and, recently, COVID-19 travel restrictions remain barriers to widespread participation. To meet this need, we instituted virtual global neurology morning reports with the objectives of (1) improving knowledge about neurologic diseases common in sub-Saharan Africa and (2) developing clinical reasoning skills through consideration of diagnostic and therapeutic limitations in resource-limited settings. Interactive case-based sessions were presented from Zambia via videoconference by a Johns Hopkins faculty member or Zambian neurology trainee. An anonymous cross-sectional survey was conducted among Johns Hopkins neurology residents. Of eligible participants, 69% (n = 30) completed the survey, 66% of whom were female, and 33% reported prior in-person global health experience. Although most participants did not anticipate a career in global health, the majority (85%) reported that exposure to global health was important. All but 1 participant (96%) reported satisfaction with the global neurology morning reports, with 100% reporting that they were useful to their clinical knowledge and 86% reporting that they were useful to their clinical practice. All respondents felt that morning reports should continue, and 69% ranked the educational value of the experience in the top quartile of the residency curriculum. Resident satisfaction with and perceived utility of global neurology morning reports were high, although the majority did not plan to pursue global neurology opportunities as part of their career. Remote learning opportunities may increase interest in global health among neurology residents.

Purpose To analyze the genetic variability in a variable number of tandem repeats (VNTR) in the thymidylate synthase (TS) enhancer promoter region and assess the influence of functional alterations in mismatch repair genes by analyzing constitutional and tumoral DNA from patients with colorectal adenocarcinoma with a high microsatellite instability (MSI-H) or microsatellite stability (MSS) status. Patients and methods Patients who underwent surgery for colorectal adenocarcinoma were selected from the colorectal database of our institute and, on the basis of MSI status, assigned to a study group and a control group: group A, MSI-H; group B, MSS. Microsatellite status was investigated using the Bethesda recommended panel (BAT-26, BAT-25, D2S123, D5S346, D17S250). In MSI-H patients an additional analysis was made of the microsatellite loci D18S61 and D18S58, both mapping in the region containing the TS gene (18p11.2–11.32). Based on the number of altered microsatellites (≥2, 1, or 0), tumors were considered as having high (MSI-H) or low (MSI-L) instability or microsatellite stability (MSS), respectively. Genotyping for thymidylate synthase promoter polymorphism was carried out on constitutional and tumor DNA of each patient by PCR amplification of the polymorphic region. Results MSI-H was found in 55 patients (group A) and MSS in 50 patients (group B). In none of the MSI-H patients was microsatellite instability found in the additional D18S61 and D18S58 loci. In five group A and ten group B cases the analysis was not performed because constitutional DNA and/or tumoral DNA were not amplifiable. Homozygotes for the triple repeat variant (3R/3R) displayed only the large PCR product, homozygotes for the double repeat variant (2R/2R) displayed only the smaller PCR product, while heterozygotes (2R/3R) displayed both the larger and smaller PCR products. In 3/50 (6%) group A patients and 5/40 (12%) group B patients repeat variations were found in tumoral DNA. Conclusion Our findings demonstrate that there is genetic homogeneity between constitutional and tumoral DNA but do not support the hypothesis that mismatch repair genes are involved in VNTR recombinant events in TS gene variability.

The aim of this paper was to compare [⁹⁰Y]-PET and SPECT imaging quantification for dosimetric applications in targeted radionuclide therapy.Imaging studies were carried out by SPECT-CT and PET equipment performing phantom tests first. [⁹⁰Y]-SPECT and PET scans were compared in terms of sensitivity, minimum detectable activity concentration, recovery coefficients (RCs) and system spatial resolution (FWHM). Quantitative evaluations by PET and SPECT acquisitions were then assessed in patients who received therapeutic activity of [⁹⁰Y]-DOTATOC directly injected into the surgical cavity by locoregional route in glioma treatment and by systemic route in neuroendocrine tumour patients who underwent intravenous infusion. Finally 3D-dose distributions by SPECT and PET images were obtained.Sensitivity was proven to be about fivefold higher for SPECT than for PET. To obtain a good-quality PET imaging, the minimum detectable activity concentration was determined to be equal to 1 MBq/mL compared with 0.05 MBq/mL that was sufficient to assess adequate SPECT imaging. RCs were 100% for volume ≥ 25.5 mL for PET and ≥ 110 mL for SPECT. FWHM was 7 mm for PET and 19 mm for SPECT scans. With regard to locoregional therapy, excellent imaging was obtained with both PET and SPECT. On the contrary, systemic administration did not permit us to obtain suitable PET imaging. PET and SPECT images were affected by considerable noise, whose influence is much more important in the quantitative evaluation of dose volume histograms rather than in the visual interpretation of images.[⁹⁰Y]-activity quantification is feasible by SPECT and PET imaging. For clinical applications, SPECT-CT is the best technique for visualizing the radiopharmaceuticals following systemic infusion, while both SPECT and PET scans are effective in analyzing locoregional distribution. Nevertheless PET study demonstrates the best spatial definition.

Ciclosporin A (CsA)-induced gingival overgrowth is attributed to an exaggerated accumulation of extracellular matrix, which is mainly due to an increased expression of transforming growth factor-β1 (TGF-β1). Herein, the in vitro investigation of effects of overexpression of Smad7, a TGF-β1 signaling inhibitor, in the events associated with CsA-induced extracellular matrix accumulation was performed.The effects of Smad7 were assessed by stable overexpression of Smad7 in fibroblasts from normal gingiva. Smad7-overexpressing cells and control cells were incubated with CsA, and synthesis of type I collagen, production and activity of MMP-2 and cellular proliferation were evaluated by ELISA, zymography, growth curve, bromodeoxyuridine incorporation assay and cell cycle analysis. The effects of CsA on cell viability and apoptosis of fibroblasts from normal gingiva were also evaluated. Western blot and immunofluorescence for phospho-Smad2 were performed to measure the activation of TGF-β1 signaling.Although the treatment with CsA stimulated TGF-β1 production in both control and Smad7-overexpressing fibroblasts, its signaling was markedly inhibited in Smad7-overexpressing cells, as revealed by low levels of phospho-Smad2. In Smad7-overexpressing cells, the effects of CsA on proliferation, synthesis of type I collagen and the production and activity of MMP-2 were significantly blocked. Smad7 overexpression blocked CsA-induced fibroblast proliferation via p27 regulation. Neither CsA nor Smad7 overexpression induced cell death.The data presented here confirm that TGF-β1 expression is related to the molecular events associated with CsA-induced gingival overgrowth and suggest that Smad7 overexpression is effective in blocking these events, including proliferation, type I collagen synthesis and MMP-2 activity.

The aim of this study is to evaluate the association between physical activity level and physical fitness with erectile dysfunction in men aged 40-75 years. We examined 180 men aged 40-75 years. The individuals were evaluated for age, presence of dyslipidemia and smoking and for anthropometric parameters for the characterization of body mass index. For assessing the level of physical fitness, a test was performed to measure the indirect maximum oxygen consumption. The evaluation of erectile function was made by the International Index of Erectile Function questionnaire and assessment of physical activity level by the International Physical Activity questionnaire in its short version. This study showed that younger men with higher physical activity and better physical fitness are less likely to suffer from erectile dysfunction. Multivariable analysis through logistic regression showed that age (odds ratio (OD)=1.15; 95% confidence interval (95% CI)=1.07-1.23), physical activity (OD=10.38; 95% CI=3.94-27.39) and physical fitness (OD=4.62; 95% CI=1.75-12.25) were independent variables associated with erectile dysfunction. This study reinforces the concept that healthy habits have a direct effect on erectile function.

This study assessed whether obesity significantly affects femoral shape. Femora of 121 white men were divided into two weight classes based on body mass index (BMI) of the deceased. Five external anteroposterior (AP) and mediolateral (ML) measurements were taken at consistent percentages of diaphyseal length. These were then subject to statistical tests. After controlling for age, multivariate statistics show a significant (p0.05) effect of BMI on the femur, with the greatest significance in ML measurements. T-tests confirm these dimensions are significantly larger in the overweight (p0.05). The effect of BMI on size-transformed and shape-transformed variables was also evaluated, with ANOVA results showing a significant BMI effect on ML size (p0.05), but not shape. Significant size-transformed ML variables were then subject to discriminate function analyses with a cross-validation correction. Results show a correct classification rate of 88% in normal weight and 77% in overweight individuals.

This report summarizes the proceedings of a workshop organized with the purpose of bringing together many of those with substantial experience in this troublesome area of pathology for an active interchange of ideas, opinions, problems, and solutions. Recognition was given the fact that current knowledge and technical capabilities are woefully inadequate for dealing with the diagnostic questions now being asked. Until such time as these inadequacies can be remedied, a very conservative approach to the interpretation of ultrastructural studies is advocated.

Chromosomal instability is increasingly appreciated as a key component of tumorigenesis in humans. A combination of abnormal telomere shortening and cell-cycle checkpoint deficiency has been proposed as the initial lesions causing destabilizing chromatin bridges in proliferative cells. We examined the participation of the different types of end-to-end fusions in generating instable karyotypes in non-transformed human breast epithelial cells. We concluded that short dysfunctional telomeres represent an initiating substrate for post-replicative rejoining of sister chromatids and are likely to make an important contribution to the formation of chromosomal rearrangements and the amplification of chromosome arm segments in breast epithelial cells. We propose that there is a chronological order in the participation of the different types of end-to-end fusions in the generation of chromosomal instability. Thus, intrachromosomal post-replicative joining would proceed mainly in the early stages after overcoming growth arrest, when telomere dysfunction is limited and affects only one chromosome end in a cell. The absence of a second substrate for end joining will conduct the cell with the uncapped chromosome to replicate its DNA and fuse the uncapped sister chromatids after replication. Later, since telomeres shorten progressively with each DNA replication round, the uncapping will affect many more chromosome ends, and fusions between the uncapped ends from different chromosomes will be produced. While the fusion of sister chromatids will produce chromosome segment amplification and terminal deletions in the daughter cells, interchromosomal fusion will produce unbalanced rearrangements other than chromosome segment amplifications.

At present, as a result of the introduction of tension-free techniques, the general opinion is that the treatment of inguinal hernia needs day-care surgery. Nevertheless day-care surgery requires a specific organization of the hospital and of the territorial services, after the discharge of the patient. These facilities are not always available all over the country. The aim of this paper is to present the results of the treatment of inguinal hernia performed by day-care surgery in an unspecialized hospital.From April 1999 to April 2000, 28 patients (25 M), median age 54 years (range 25-85), affected by inguinal hernia, underwent ernioplasty operation according to Trabucco technique, by only one skilled surgeon, in an unspecialized hospital. Five patients were affected by inguinal hernia of type I of Nyhus, 9 of type II, 10 of type IIIa and 4 of type IIIb. Twenty-five patients underwent spinal anaesthesia and 3 general anaesthesia due to failure of the previous one or for contraindications due to spine diseases or due to patient's request. All patients remained in the hospital the night after the operation. After discharge, patients were controlled in the outpatient ambulatory every other day for 10 days. After that, patients were required to submit to physical examination only if they were symptomatic. At present, average follow-up is of 18.1+/-6.3 months.The following unspecific complications prevalently due to spinal anaesthesia were observed: 2 (7.1%) acute urinary retentions, 1 (3.6%) headache, 1 (3.6%) acute hypotension, 2 (7.1%) feverets, 1 (3.6%) vomiting, and 1 (3.6%) influenza. The specific complications were: 2 (7.1%) transient neuralgias, 2 (7.1%) ecchimosis and 1 (3.6%) infection of the wound. Average intensity of postoperative pain (VAS) was of 2.8+/-1.2. The average abstention from work was of 12.5 days. Until now, any recurrence has been observed.In spite of the few cases, our results are similar to those of specialized hospitals organized for day-care surgery. Anyway, in the management of these patients in unspecialized hospitals, according to our opinion one night in the hospital after the operation is required for the easiness of the patient and of the surgeon.

Alagille syndrome (arteriohepatic dysplasia) is a major cause of intrahepatic cholestasis in infancy. The present report describes a patient with Alagille syndrome who presented with hematuria and IgA nephritis 7 years after an orthotopic liver transplantation and immunosuppression. This patient suggests that glomerular lipidosis is not an inherent feature of the Alagille syndrome, and that IgA nephritis may develop in spite of ongoing immunosuppressive treatment.

The syndrome of resistance to thyroid hormone is associated with diverse mutations in the ligand-binding domain of the thyroid hormone beta receptor, localizing to three clusters around the hormone binding cavity. Here, we report three novel resistance to thyroid hormone mutations (S314C, S314F, and S314Y), due to different nucleotide substitutions in the same codon, occurring in six separate families. Functional characterization of these mutant receptors showed marked differences in their properties. S314F and S314Y receptor mutants exhibited significant transcriptional impairment in keeping with negligible ligand binding and were potent dominant negative inhibitors of wild-type receptor action. In contrast, the S314C mutant bound ligand with reduced affinity, such that its functional impairment and dominant negative activity manifest at low concentrations of thyroid hormone, but are more reversible at higher T3 concentrations. The degree of functional impairment of mutant receptors in vitro may correlate with the magnitude of thyroid dysfunction in vivo. Modelling these mutations using the crystal structure of thyroid hormone receptor beta shows why ligand binding is perturbed and why the phenylalanine/tyrosine mutations are more deleterious than cysteine.

The site of leakage in a patients with rhinorrhea of various origin may be difficult to identify. The aim of our paper is to evaluate the contribution of cisternography in combination with single photon emission tomography (SPECT) to identify the fistulous track. From 1/1/1992 to 30/11/1997 we studied 20 patients with rhinorrhea posing a challanging diagnostic problem as to identification of the leakage site. Two mls of Indium DTPA (In 111) were injected into the subarachnoid space by the lumbar route. The tracer was followed by planar scintigraphy until it reached the cranial base and subsequently the SPECT acquisition started. A fistula was demonstrated in all of our cases including patients with no active leakage at the time of examination, patients with no bone defects on thin sliced CT scanning or patients with a normal MRI. At surgery the fistulous track was confirmed in all but two cases when a bilateral fistula was operatively identified only on one side. In conclusion whenever a CT scanning fails to demonstrate significant bone defects and MRI does not localize a fistulous track, SPECT cisternography via the lumbar route proved in our experience to be a reliable examination for a precise diagnosis.

Resistance to thyroid hormone (RTH) is almost invariably associated with mutations of the thyroid hormone (TH) receptor beta (hTR beta) gene and is inherited as an autosomal dominant disease. Mutations of hTR beta identified in patients affected by RTH cluster generally at two spots of the ligand binding domain. We investigated whether an Italian kindred with RTH had a mutation in the thyroid hormone (TH) receptor beta gene. Blood samples were obtained from the available family members for biochemical and genetic analyses. Thyroid function tests in basal conditions, and in the case of the propositus also following incremental doses of T3, were performed. Exon 4 to 10 of hTR beta gene were amplified using the polymerase chain reaction (PCR) and the mutation was identified by direct sequence analysis. The affinity constant of this mutated receptor for T3 was measured by in vitro transcription-translation and was then compared with that of wild type. We identified a heterozygous G to A transition at nucleotide 1037 of exon 8 at codon 251, resulting in a glycine (G) to glutamic acid (E) substitution (G251E) in the patient affected by RTH and in his affected offspring, but not in the normal family members. This novel mutation represents a de novo mutation since both parents of the index case were unaffected and did not have this genomic mutation. When expressed in vitro, the mutant protein (G251E) showed a marked decrease of the affinity for T3, suggesting an impaired ligand-dependent transactivation activity of this mutant receptor. In vivo studies with incremental doses of L-T3 demonstrated a reduced sensitivity to TH in the index case, in particular at the pituitary level where the thyrotrophs' activity was not completely inhibited even by 200 micrograms/day of L-T3. G251E mutation represents the fourth mutation described up to now in exon 8 of hTR beta among the subjects affected by RTH. A third cluster of mutations of the c-erbA beta gene located proximally with respect to the other two so far described begins to emerge in RTH patients.

The authors report a case of urachal fistula diagnosed after repeated omphalitis, this disease was associated with coronal hypospadias. Diagnosis was done thanks to a fistulography; subsequently, during the operation, to evidence the fistula, this was injected with methylene blue, this expedient showed that the fistula was longer than that disclosed from fistulography. To carry out a complete extirpation of the fistula the authors were bound to make a cystotomy. The course after operation was good and the patient discharged after a few days. After 5 years the patient is in perfect health. This case allowed some anatomic, clinical and semeiotic considerations.

Byler disease (ByD) is an autosomal recessive disorder in which cholestasis of onset in infancy leads to hepatic fibrosis and death. Children who have a clinically similar disorder, but are not members of the Amish kindred in which ByD was described, are said to have Byler syndrome (ByS). Controversy exists as to whether ByD and ByS (subtypes of progressive familial intrahepatic cholestasis [PFIC]) represent one clinicopathological entity. The gene for ByD has been mapped to a 19-cM region of 18q21-q22. PFIC caused by a lesion in this region, including ByD, can be designated PFIC-1. Examination of haplotypes in siblings with ByS in two unrelated non-Amish families showed that the gene(s) responsible for their disorder(s) did not lie in the PFIC-1 candidate region. On light microscopy and transmission electron microscopy (TEM), liver tissue differed between Amish children with PFIC-1, who had coarsely granular bile and at presentation had bland intracanalicular cholestasis, and the children with ByS in the two non-Amish families, who had amorphous or finely filamentous bile and at presentation had neonatal hepatitis. Bile acid composition of bile also differed: In the Amish children with PFIC-1 and in one ByS family, the proportional concentration of chenodeoxycholic acid (CDCA) in bile was low compared with normal bile; in the other ByS family, it was only slightly reduced. Genetic analysis and light microscopy and TEM of liver may help distinguish PFIC-1 from other forms of ByS.

The whole male population with indwelling catheter of the Trento health district has been included in a survey of the quality of the home nursing services. The epidemiological and in depth analysis of the characteristics and motivation of the high degree of heterogeneity of medical prescribing behaviors and of the nursing practices suggest that an important effort of rationalization (on the basis of pre adopted protocols) is both needed and possible.

The authors describe 9 cases of rhabdomyolytic acute renal failure (ARF) as a complication of cardiopulmonary bypass.Retrospective research between June 1992 and March 1994.Department of Cardiac Surgery.931 consecutive patients undergoing major cardiac surgery.Patients affected by rhabdomyolytic ARF were treated with pharmacological therapy and/or plasmapheresis/continuous arteriovenous hemofiltration. In seven patients indirect cannulation of the femoral artery was used.Incidence, risk factors of syndrome results obtained with pharmacological treatment, CAVH and plasmapheresis were evaluated. Statistical analysis was performed with ANOVA, Tukey Kramer test and chi2 test (p0.05 as significant).The syndrome occurred in 0.96% (9/931 patients) of the total cases; 11.3% (6/53 -p0.0000) in patients undergoing a direct femoral artery cannulation for cardiopulmonary bypass and 9.5% (2/21, p0.01) in patients in which the aortic balloon pump was used. Six patients develop acute anuric renal failure and underwent plasma exchange and hemodialysis (1 case) or CAVH (5 cases); 3 patients underwent early medical treatment and developed developed acute renal failure (ARF) with preserved diuresis. Early medical therapy appeared to prevent the evolution towards anuric ARF. The indirect cannulation of the femoral artery does not seem to produce a rhabdomyolytic ARF syndrome. In patients with direct femoral artery cannulation risk factors appear to be: arteriopathy (p0.001), prolonged extra corporeal circulation (p0.001), low cardiac output syndrome (p0.001), continuous i.v. infusion of epinephrine (p0.0001).Rhabdomyolytic acute renal failure is a severe complication, early identification of patients ¿at risk¿ is most important. The preventive measures and the therapy adopted proved efficient.

An uncommon case of traumatic rupture of chordae tendineae with tricuspid regurgitation is described. The early diagnosis, in a patient with polydistrectual injuries, was done by transesophageal echocardiography. This technique allowed accurate diagnosis of the type of lesion suggesting the opportunity to perform an echocardiogram, possibly transesophageal, in all patients with blunt thoracic trauma, even in the absence or with subtle clinical manifestations of cardiovascular involvement.

In a controlled, randomized study, we evaluated duration of neuromuscular block in 80 patients undergoing routine abdominal surgery. Forty patients were aged 18–50 yr (control group) and 40 patients were more than 65 yr (elderly group). All patients had normal plasma creatinine concentrations. After induction of anaesthesia, patients were allocated randomly to receive either atracurium 0.5 mg kg−1 or vecuronium 0.1 mg kg−1 to facilitate tracheal intubation. Monitoring of the evoked response of the adductor pollicis muscle to supramaximal single twitch ulnar nerve stimulation every 10 s was performed and measured with a strain gauge. Repeat doses of atracurium 0.1 mg kg−1 or vecuronium 0.02 mg kg−1 were administered when the adductor pollicis response recovered to 25% of the control twitch height. We found that the duration of action of the initial dose of atracurium was similar in the control and elderly groups, and it did not vary after repeated doses. However, the initial dose of vecuronium caused a significantly longer period of clinical block in the elderly group compared with the controls, and the duration of action of repeated doses was longer in the elderly group. We conclude that as there is a risk of prolonged effect of vecuronium in the elderly, monitoring of neuromuscular function is recommended in this group. Alternatively, atracurium should be preferred for prolonged surgery in elderly patients.

Postischemic septal rupture has always been evaluated, in respect of surgical indication, as regards the time lapse between infarct and rupture, interval between rupture and operation, extension of myocardial damage and general risk factors such as age, sex and associated pathologies. But in fact the surgeon is dealing with a two sided problem, the MI and the rupture, and thus surgical results depend upon both the residual ventricular function after MI and the consequences of volume overload on a damaged muscle. Surgical indication could not be based on a single criterion only. Extension of the MI alone is not fully predictive of operative mortality because, aside the reperfusion injury, the repair further jeopardizes viable myocardium and alters ventricular geometry; although the shunt appears unrelated to mortality it certainly interferes with operative outcome at least because of the time elapsed between rupture and repair. So far an index which could correlate the extension of myocardial damage and the entity of the shunt with each other was not available. Patients with septal rupture follow an emergency protocol and are often insufficiently investigated but every patients has a least one echo-Doppler evaluation or even a ventriculography while one or more ECGs are always available. With the presumption that the Qp/Qs is roughly indicative of the right ventricular volume overload and that ecg signs of myocardial infarct are always reliable, we have reviewed among our 24 patients with septal rupture those where a full ecg tracing and a quantitative Echo or angiographic evaluation of the shunt were available.(ABSTRACT TRUNCATED AT 250 WORDS)

DA/DAPI and C chromosomal heteromorphic sites (1q, 9q, 15p, 16q, Yq) in a sample of 136 unrelated individuals from the Garfagnana valley (Tuscany, Italy) have been analyzed quantitatively and qualitatively. The variations in length, between-homolog heteromorphisms, and intensity of fluorescence of the heterochromatic bands have been compared in two subsamples of the population (upper and middle valley) individualized according to geodemographic criteria. DA/DAPI heterochromatin differed significantly from C heterochromatin, showing a lower average amount and a higher variability at each site. This suggests a differential staining of DNA of the two banding systems. Furthermore, DA/DAPI heterochromatin was less uniformly distributed in the population than C heterochromatin and the regions 16q and Yq discriminated better between subsamples. The variations of DA/DAPI fluorescence at the 15p site demonstrated an excess of homomorphic individuals in the upper valley, which could be related to the mating structure of the population living in this area.

Blood group systems ABO, KEL, MN, RH, secretor status for ABH blood group substances, red cell enzymes ACP1, ESD, AK1, 6-PGD, PGM1 subtypes, serum markers GC, TF, PI, AHSG, ORM1 and chromosomal heteromorphism Q, C, DA/DAPI were examined in a population sample of the Garfagnana, a semi-isolated mountainous area in the province of Lucca (Tuscany, Italy). The total sample was subdivided and analysed according to the more recent historical events which have occurred in Garfagnana. The observed phenotype and gene frequencies were discussed in the context of other population samples from the same province and of the same region. As a whole, the results concur in pointing out a certain degree of heterogeneity within the area and a high differentiation from the rest of Tuscany. Such peculiar genetic features of the population have to be related to the historical, geographic and cultural isolation of the Garfagnana.

A 3-year-old girl of Navajo heritage had intractable diarrhea beginning at 4 days of age and resulting in long-term hyperalimentation. Investigation before multivisceral transplantation included biopsies of the rectum, stomach, duodenum, and liver. The diagnosis of microvillus inclusion disease was established by documentation of microvillus inclusions in duodenal epithelial cells. A trial of somatostatin therapy was ineffective in controlling the diarrhea. Subsequently, a multivisceral organ transplant provided a unique opportunity to establish the gastrointestinal extent of involvement of this disease. Ultrastructural microvillus inclusions were identified in the duodenum, jejunum, ileum, and colon, but not in the gallbladder. A few inclusions also were documented in gastric antral epithelial cells. Alkaline phosphatase stains performed on paraffin-embedded material showed a few inclusions in the antrum of the stomach and many inclusions throughout the small intestine, primarily in surface epithelial cells but also in upper crypt cells.

The Authors report a case of peritonitis due to perforation of a single small bowel metastasis in a woman affected by right upper lobar lung carcinoma still with neck's lymphnodes and cerebellar metastasis. They underline the rarity of the finding and analyze its clinical, therapeutic, prognostic problems.

Post-infarction free-wall, papillary muscle and septal myocardium rupture yields a lowering mortality, not only due to the improved surgical technique but also to a more careful selection of surgical candidates. The value of pericardial decompression in cases of free-wall rupture is discussed. Echocardiographic evidence of blood in the pericardium after a myocardial infarction is not a direct indication for a diagnostic sampling, decompression or surgery. Pericardiocentesis is not a risk-free procedure and should be limited to patients with life-threatening acute tamponade. Clinical results of 34 patients operated upon for myocardial rupture, regardless of adopted technique or timing of surgery, confirm that the decision about a patient's operability should follow a careful evaluation of the following risk factors in this order: infarct size, age, cardiogenic shock, multiorgan failure, rupture site, cardiac failure and associated diseases.

Ten patients with advanced ovarian carcinoma (5 in FIGO stage III and 5 in stage IV), following the failure of repeated chemotherapy courses, were submitted to i.p. radioimmunotherapy (RIT). Several antibodies (HMFG1, HMFG2, H17E2, B72.3) were employed. The antibody choice was made on the basis both of immunohistochemistry and immunoscintigraphy results. The mean dose administered in each single course was 17 mg, 3 patients underwent two treatments and 2 patients were injected three times. The isotope employed was 131I (mean dose 92 mCi). The early or late untoward effects were negligible. All patients developed anti-mouse antibodies (HAMA) which reduced, during the further RIT, the MoAbs' effective half-life. The estimated radiation dose delivered to the tumour ranged from 2910 to 11,000 cGy. Both wholebody and liver radiation burden resulted in tolerable limits while the thyroid irradiation was high. In 3 patients the RIT result was ineffective; in 2 cases a temporary ascitis staunch was obtained. In 2 cases the tumour growth was slower than expected and in 3 patients instrumental evidence of partial tumour reduction, still lasting after 4–11 months, was observed.

Type IV glycogenosis is due to branching enzyme deficiency and is usually manifested clinically by progressive liver disease with cirrhosis and hepatic failure between the second and fourth years of life. We describe a 5-year-old boy who, following an acute febrile illness at 2 years of age, was first noted to have hepatomegaly with mildly elevated serum transaminase levels. Liver biopsy revealed hepatic fibrosis with periodic-acid Schiff-positive, diastase-resistant inclusions in hepatocytes and fibrillar inclusions characteristic of amylopectin by electron microscopy. Enzymatic assay revealed deficient hepatic branching enzyme activity with normal activity of glucose-6-phosphatase, debranching enzyme and phosphorylase activities. During the succeeding 3 years, he grew and developed normally with apparent resolution of any clinical evidence of liver disease and only intermittent elevation in serum transaminase levels associated with fever and prolonged fasting. Repeat liver biopsy at 4 years of age showed persistence of scattered hepatocellular periodic-acid Schiff-positive, diastase-resistant inclusions, but no progression of hepatic fibrosis in spite of persistent deficiency of hepatic branching enzyme activity. Skeletal muscle and skin fibroblasts from the patient also showed deficient enzyme activity. Skin fibroblasts from both parents exhibited half the normal control activity, suggesting a heterozygote state. This is the first documented patient with deficiency of branching enzyme but without evidence of progressive hepatic disease. This patient, coupled with reports of other patients with late onset hepatic or muscle disease with branching enzyme deficiency, suggests that the defect resulting in Type IV glycogen storage disease is more heterogenous and possibly more common than previously suspected.

Plasma levels of lactoferrin (LF) have been found to be increased in a few patients with cystic fibrosis (CF). This study was aimed at investigating plasma LF levels in children with CF (26 cases) and in controls (C) (19 cases). Plasma LF was measured by a radioimmunoassay method. Plasma LF levels were not significantly different in CF and in C, even though 10 CF patients showed LF levels above the mean + 2 SD value of the controls. Neither the duration of the disease nor the age of the controls was correlated with LF or with the exocrine pancreatic capacity. A significant relationship between the presence of an acute lung inflammation and LF levels was found. This study shows that LF is increased in CF only in the presence of an acute inflammatory state. Further studies are necessary to establish the usefulness of an LF assay as an index of the presence of an acute inflammatory process.

SUMMARY Single-dose pharmacokinetics of azlocillin, cefoperazone and ceftazidime were studied in 17 patients with cystic fibrosis (CF). All patients had broncho-pulmonary infections caused by Pseudomonas aeruginosa. Three groups of five, six, and six patients were treated with azlocillin, cefoperazone, or ceftazidime, respectively. The size of the single dose was 133mg/kg for azlocillin, 66u.7mg/kg for ceftazidime and 66.7 mg/kg for cefoperazone. The clearance values for the three antibiotics calculated from the single-dose data were, on the average, higher than the values previously reported for normal subjects. After the first dose, the patients received a repeated-dose treatment with the same antibiotic. During the first 5 days of therapy, a complete postural drainage of sputum was obtained four times a day for each patient. Cefoperazone could be measured in 47 (39.2%) of the 120 sputum samples assayed while ceftazidime was shown to be present in all 120 sputum samples examined. Azlocillin was not detected in any of the 100 sputum samples assayed.

Based on a survey of 47 cases of left colonic occlusive cancer operated on at the Emergency Surgery Department-University of Rome "La Sapienza", the authors concluded that in such condition, when the patient is not seriously ill, the Hartmann procedure is a valid alternative to the anastomosis-resection because: a) it solves intestinal obstruction in a short time with consequent improvement of general conditions; b) it allows to perform the second oncologically radical operation early with less risk of neoplastic spread. The latter procedure, therefore, is carried out under better conditions, relative to the stage of the disease and the status of the patient.

Thirteen patients with a diagnosis of primary colorectal tumour and 68 patients previously operated for colon cancer underwent an immunoscintigraphy carried out with monoclonal antibodies anti-CEA F(ab')2, labelled with 131I or 111In. These studies led to the detection of all primary tumours and of most of their associated lesions (7/8). In this group 15 neoplastic deposits previously undetected were demonstrated, allowing an improvement in the patient staging before the operation. In the group of the 68 operated patients, immunoscintigraphy succeeded in imaging 96 out of 121 known lesions: the best outcomes were obtained for abdominal and pelvic recurrences as well as for lymph nodes lesions. The lowest levels of sensitivity were observed for liver metastases. An enhancement of the immunoscintigraphy sensitivity was obtained when the radiopharmaceutical was injected intraperitoneally. In 50 patients, this alternative administration route was successful in detecting 91/107 cancer deposits and, in particular, liver metastases were found out in 36/42 cases.

Six human T lymphocyte clones, characterized as to cytolytic activities and several surface markers, were investigated for possible morphologic correlates of function. Cytotoxic T lymphocyte clones and natural killer-like clones had basically similar morphology, but cytotoxic T lymphocytes tended to cluster in groups and had many more lipid bodies, whereas natural killer-like cells had fewer intercellular contacts and had more and larger electron-dense bodies. Dense bodies were also quite prominent in noncytotoxic clones. The latter were distinctive in having a minority of very large cells (10 to 16 microns) with many microvilli, scattered among the majority of 5- to 8-microns diameter cells.

A light- and electron-microscopic review of 40 cases diagnosed at Children's Hospital of Pittsburgh as ganglioneuroblastoma, neuroblastoma, or small round cell tumor-probably neuroblastoma disclosed four cases that contained abundant glycogen. Two were unquestionable neuroblastomas by electron microscopy; one was primary in the adrenal gland, the other in the mediastinum. In the third case, a paraspinal tumor, the light-microscopic appearance was suggestive or neuroblastoma, but no catecholamine granules or neural processes were demonstrated in the material available for electron microscopy. The fourth case was an undifferentiated malignant tumor in the pectoralis muscle of a 12-year-old girl. By electron microscopy, neural processes were demonstrated and the tumor was classified as peripheral neuroblastoma. Of the remaining 36 cases, electron microscopy readily indicated a diagnosis of neuroblastoma or glangioneuroblastoma in 35 of them. In the other case, the tissue had been fixed in formalin and only a few catecholamine granules were found after an extensive search.

Technetium-99m and/or 111In labelled F(ab')2 fragments of a melanoma associated MoAb 225.28S were injected i.v. in 80 patients affected by stage I to IV malignant melanoma. Seventy five percent of metastatic lesions already documented by other methods were detected by immunoscintigraphy, which was also capable of detecting a certain number of unknown metastases. However, we observed a lower percentage of positive scans in liver, lung and skin because of the poor tumour to background ratio. In some patients, subcutaneous (s.c.) injection allowed us to visualize documented metastases undetected by i.v. administration. An equal amount of non-specific F(ab')2 fragments (MoAb 4C4) injected s.c. as a negative control showed no positive scans. Clinical studies and chromatographic patterns of patient serum samples suggest that the s.c. route of administration offers, with respect to the i.v. route, the advantage of reducing vascular background and aspecific accumulation in liver, probably because of retention of possible contaminants by the lymphatic system.

We have investigated the effectiveness of seven new beta-lactam antibiotics, azlocillin, piperacillin, ceftazidime, cefsulodin, cefoperazone, latamoxef (moxalactam), and cefotaxime, against acute pulmonary exacerbations caused by Pseudomonas aeruginosa in cystic fibrosis. Three hundred and fifty-five strains of Ps aeruginosa isolated from 310 sputum cultures (190 cystic fibrosis patients) were tested for susceptibility to the drugs by determination of minimal inhibitory concentrations (MIC). The highest activity was shown by ceftazidime (6% resistant strains) followed by cefsulodin and piperacillin (15 and 16% resistant strains); very low activity was found for cefotaxime and latamoxef (moxalactam). Ceftazidime was the most active drug against 32 pseudomonas isolates that were resistant to both carbenicillin and aminoglycosides (78% susceptible). A randomized, double-blind trial of azlocillin, piperacillin, ceftazidime, cefsulodin or cefoperazone was performed in 111 cystic fibrosis patients with predominant and susceptible pseudomonas in their sputum. Results were evaluated by a clinical, radiological and bacteriological scoring system: the best results were obtained with ceftazidime, followed by cefsulodin and piperacillin. However, pseudomonas was eradicated in only 22 (23%) of the cases with the most active drugs and persisted or reappeared in all the cases 1 to 3 months later. Ceftazidime always eradicated Staph. aureus and Haemophilus influenzae associated with pseudomonas. Similar eradication occurred nearly always with cefsulodin but rarely with the other drugs. No serious drug reaction occurred but a later fever and rash with piperacillin, transient diarrhoea with cefoperazone, vomiting with cefsulodin, and very frequent eosinophilia with ceftazidime should be mentioned. These five drugs offer, in varying degree, alternatives to traditional anti pseudomonas antibiotics in cystic fibrosis pulmonary infections, but they should be used only against well-proven resistant strains. Ceftazidime is best and cefotaxime and latamoxef (moxalactam) least useful.

Despite sufficient evidence to the contrary, fibrillar nuclei continue to be claimed by some to represent paramyxovirus. In a review of the electron-microscopic material, fibrillar nuclei were found in a variety of tissues and situations where a viral etiology is unlikely. Fibrillar nuclei were most often found in postmortem and formalin-fixed material. These nuclei were also experimentally produced in postmortem human lung with formalin fixation followed by a deionized water rinse. It is concluded that fibrillar nuclei do not represent virus, but chromatin, and it is believed that this chromatin appearance is related to cell injury and to tissue processing. It is also believed that fibrillar nuclei occur with much greater frequency than realized. These nuclei are usually ignored during examination of specimens by light and electron microscopy because these specimens are usually being selectively screened for other changes and perhaps because a clear understanding of their significance is still lacking.

The clinical and pathologic findings of two patients with malignant ectomesenchymomas of soft tissue are reported. Malignant ectomesenchymomas are a composite of ganglion cells or neuroblasts and one or more malignant mesenchymal elements, usually rhabdomyosarcoma. Our first case was composed of ganglioneurosarcoma plus rhabdomyosarcoma, the second composed of neuroblastoma plus rhabdomyosarcoma. The name is derived from the suggestion that they arise from pluripotential ectomesenchyme. An English language literature search revealed 11 other cases that arose in soft tissue and had adequate clinicopathologic data. Of the 13 cases, 10 occurred in infants, three occurred in adults, and nine were males. Six patients were alive and free of disease at last follow-up (range, 0.6-12 years; mean, 3.4 years), four patients had died of tumor (within 0.5-3.3 years; mean, 1.3 years), one case had died of Adriamycin (doxorubicin) toxicity, and two cases had been lost to follow-up. Complete surgical resection is the mainstay of treatment and chemotherapy appears to be important.

Four small round cell malignant tumors of bone occurring in children are described. There was no catecholamine secretion and the clinical, radiologic, and biopsy diagnosis in each was Ewing's sarcoma. Glycogen was sparse both on imprints and in tissue sections. The tumors, when extensively sampled, had areas of a lobular growth pattern and Homer Wright rosettes. The rosettes were always focal and varied in complexity from case to case; they were rudimentary in one instance and markedly fibrillar in the most obvious instance. Neuron-specific enolase was demonstrated in tissue sections and in longterm cell cultures from three of the tumors. The cultured cells put out moderately long beaded processes in serum-free medium but had no catecholamine fluorescence. Electron microscopy of the tumor rosettes and the cultured cells showed processes containing aggregates of microtubules and only one case had rare neurosecretory granules. This study suggests that some small round cell tumors of bone and soft tissue in children, which present as Ewing's sarcoma, are neuroectodermal in nature.