In-vitro infection of chronic lymphocytic leukemia cells by Epstein-Barr virus (EBV)

We sought to determine the potential of infecting lymphoid cells from patients with chronic leukemia (CLL) with Epstein-Barr virus (EBV) by testing for EBV receptors (EBVR) by flow cytometry, assessing for infectability of these cells by culturing with B95-8-derived virus, and staining for EB nuclear-associated antigens (EBNA) at various times post-infection. EBVR were present on 54-91% of lymphoid cells in seven cases of CLL and on 46% of prolymphocytic leukemia cells. Dynamic changes regarding EBNA positivity, morphology, and viability occurred post-infection with the virus. On day 2 only a few EBNA-positive lymphoblasts were observed. On days 11-21 positivity increased from 2 to 34% of cells. Simultaneously, the viable cell number declined to approximately 1/10th of original number. A significant proportion of the EBNA-positive cells corresponded to the original CLL cells. In 3 of 7 cases of CLL a Pan T-cell phenotype was demonstrated by Leu-1 monoclonal antibody testing. The infected cells did not react with two monoclonal antibodies, EBV-CS 1 and 4, which react with B-cell lymphoblastoid cell lines (B-LCL). Moreover, the B-LCL derived at 1-2 months post-infection of CLL cells did not express the Leu-1 antigen, but expressed EBV-CS 1 or 4 defined antigens. In the prolymphocytic leukemia, 64% of the cells showed EBNA positivity on day 7 and giant cells with huge round or multiple nuclei appeared which were EBNA-positive. CLL and prolymphocytic leukemia cells can be infected as demonstrated by EBNA-positivity. This infection does not lead to immediate transformation, but evokes lymphoblast and multinucleated giant cell production prior to the death of cells.     

Treatment of substance dependence by a bio-cognitive model based on behavioral pharmacology]

We have introduced cognitive behavior therapy (CBT) into the treatment of substance dependence patients, which involves disease education and focused group therapy to obtain insight into the taking behavior and to establish concrete countermeasures to prevent relapse. We have created a bio-cognitive model based on biological aspects to explain the pathology of substance dependence. 'Dependence' is a term in behavioral pharmacology defined as reinforced drug seeking and taking behavior. Changes in taking behavior are thought to occur due to the repetition of the reinforcement action of psychoactive substances in the reward system of the brain. Therefore, when intake desire is strong, it is hard for patients to control themselves, and there is a feature of difficulties considering the process of thinking in CBT. In other words, when craving becomes strong, a chain of behavior happens spontaneously, without schema, involving automatic thoughts. We think that the improvement of protracted withdrawal syndrome (PWS) and entire frontal lobe function are important in learning to discern distortion of cognition. When PWS is improved, a conflict is easy to bring about in the process of drug seeking and taking behavior. And, it is easy to execute avoidance plans (coping skills) which are established to cope with craving in advance. We think that a goal for treatment is to discern drug seeking and taking behavior with natural emotion. The recovery of PWS and frontal lobe dysfunction takes a long time with a serious dependence, so we must perform repetition of CBT. As the treatment introduction of involuntary admission cases is adequate or cases of 1 to 3 months of admission treatment based on voluntary admission are hard to treat, treatment to obtain insights into patients while carrying out repeated CBT using a bio-cognitive model and to improve PWS could be a possibility as one treatment for the pathology of diversified substance dependence.     

Implantation treatment method of slow release anticancer doxorubicin containing hydroxyapatite (DOX-HAP) complex. A basic study of a new treatment for hepatic cancer.

We performed an experimental study on slow releasing anticancer drug implantation treatment as a new therapy for hepatocellular carcinoma. Hydroxyapatite (HAP) was chosen for the carrier material and doxorubicin hydrochloride (DOX) for anticancer agent. DOX-HAP was produced by adsorbing DOX to porous HAP particles of 1375 +/- 125 microns diameter using the freeze drying method. In vitro experiments showed slow release of the drug resulting in the steady release of DOX from HAP for 1 month duration. In healthy white rabbits with DOX-HAP implantation in the liver, serum DOX was not detectable, and DOX release rate was stable at the implanted region after 7, 14, and 21 days. When DOX-HAP (DOX; 100 mg kg-1) was administered to mice with sarcoma 180, an improved survival rate was observed without acute toxicity. We also found that VX2 liver tumour growth on white rabbit was inhibited by implantation of DOX-HAP, without acute toxicity. We hope that DOX-HAP implantation therapy will open up new avenues for the treatment of hepatoma.     

ADULT HEPATOBLASTOMA: CASE REPORT AND REVIEW OF THE LITERATURE

We report a case of hepatoblastoma in a 24 year old woman. The tumour was removed by an extended right lobectomy after transcatheter hepatic arterial embolization. A survey of the literature revealed 31 cases of adult hepatoblastoma. According to these reports there are no characteristic clinical features of the tumour, making a correct diagnosis before surgery or autopsy extremely difficult. Complete removal of the tumour is the only treatment which is potentially curative provided that lesion is confined within the liver capsule.     

Congenital arhinia: molecular-genetic analysis of five patients

Congenital arhinia, complete absence of the nose, is an extremely rare anomaly with unknown cause. To our knowledge, a total of 36 cases have been reported, but there has been no molecular-genetic study on this anomaly. We encountered a sporadic case of congenital arhinia associated with a de novo chromosomal translocation, t(3;12)(q13.2;p11.2). This led us to analyze the patient by BAC-based FISH for translocation breakpoints and whole-genome array CGH for other possible deletions/duplications in the genome. We found in this patient an approximately 19 Mb deletion spanning from 3q11.2 to 3q13.31 but no disruption of any gene(s) at the other breakpoint, 12p11.2. As the deleted segment at 3q was a strong candidate region containing the putative arhinia gene, we also performed the array CGH in four other arhinia patients with normal karyotypes, as well as mutation analysis of two genes, COL8A1 and CPOX, selected among hundreds of genes located to the deleted region, because they are expressed during early stages of human craniofacial development. However, in the four patients, there were no copy number aberrations in the region examined or no mutations in the two genes. Although our study failed to identify the putative arhinia gene, the data may become a clue to unravel the underlying mechanism of arhinia.     

Dose-effect relationships between manganese exposure and neurological, neuropsychological and pulmonary function in confined space bridge welders

Background

Although adverse neuropsychological and neurological health effects are well known among workers with high manganese (Mn) exposures in mining, ore‐processing and ferroalloy production, the risks among welders with lower exposures are less well understood.

Methods

Confined space welding in construction of a new span of the San Francisco–Oakland Bay Bridge without adequate protection was studied using a multidisciplinary method to identify the dose–effect relationship between adverse health effects and Mn in air or whole blood. Bridge welders (n = 43) with little or no personal protection equipment and exposed to a welding fume containing Mn, were administered neurological, neuropsychological, neurophysiological and pulmonary tests. Outcome variables were analysed in relation to whole blood Mn (MnB) and a Cumulative Exposure Index (CEI) based on Mn‐air, duration and type of welding. Welders performed a mean of 16.5 months of welding on the bridge, were on average 43.8 years of age and had on average 12.6 years of education.

Results

The mean time weighted average of Mn‐air ranged from 0.11–0.46 mg/m³ (55% >0.20 mg/m³). MnB >10 µg/l was found in 43% of the workers, but the concentrations of Mn in urine, lead in blood and copper and iron in plasma were normal. Forced expiratory volume at 1s: forced vital capacity ratios (FEV₁/FVC) were found to be abnormal in 33.3% of the welders after about 1.5 years of welding at the bridge. Mean scores of bradykinesia and Unified Parkinson Disease Rating Scale exceeded 4 and 6, respectively. Computer assisted tremor analysis system hand tremor and body sway tests, and University of Pennsylvania Smell Identification Test showed impairment in 38.5/61.5, 51.4 and 88% of the welders, respectively. Significant inverse dose–effect relationships with CEI and/or MnB were found for IQ (p⩽0.05), executive function (p⩽0.03), sustaining concentration and sequencing (p⩽0.04), verbal learning (p⩽0.01), working (p⩽0.04) and immediate memory (p⩽0.02), even when adjusted for demographics and years of welding before Bay Bridge. Symptoms reported by the welders while working were: tremors (41.9%); numbness (60.5%); excessive fatigue (65.1%); sleep disturbance (79.1%); sexual dysfunction (58.1%); toxic hallucinations (18.6%); depression (53.5%); and anxiety (39.5%). Dose–effect associations between CEI and sexual function (p<0.05), fatigue (p<0.05), depression (p<0.01) and headache (p<0.05) were statistically significant.

Conclusions

Confined space welding was shown to be associated with neurological, neuropsychological and pulmonary adverse health effects. A careful enquiry of occupational histories is recommended for all welders presenting with neurological or pulmonary complaints, and a more stringent prevention strategy should be considered for Mn exposure due to inhalation of welding fume.Manganese (Mn) exposure through welding fume has been reported to cause parkinsonian syndrome, sometimes described as welding fume‐related parkinsonism and often misdiagnosed as Parkinson''s disease because of similarities in neurological features including tremor, masked facies and generalised bradykinesia.¹ However, important differences in movement disturbance between Mn‐induced parkinsonism (also called manganism) and idiopathic Parkinson''s disease (IPD) include an awkward high‐stepping dystonic gait in manganism, which is in stark contrast with the typical shuffling gait in patients with IPD, and the tendency to lose balance by falling backward in IPD and forward in manganism. Additionally, tremor characteristics are different, usually resting tremor in IPD and postural intention tremor in manganism.¹ Magnetic resonance imaging (MRI) may be used to differentiate IPD from Mn‐induced parkinsonism if a patient has had excess Mn exposure within the previous 6 months.² Positive MRIs can show cerebral Mn²⁺ deposition in both animals and humans, especially in the globus pallidus (possibly in the striatum), by exhibiting a T₁‐weighted signal hyperintensity.³ IPD, however, is associated with lesions in the substantia nigra pars compacta, and does not exhibit MRI abnormality in the globus pallidus. Two other key features differentiating between Mn‐induced parkinsonism and IPD are: (1) a younger age of onset for Mn‐exposed workers and (2) little or no response to l‐dopa among Mn‐induced parkinsonism cases.^4,5 Case reports of neurological findings in career welders exposed to Mn have shown dystonia bilaterally in the shoulders and four distal limbs, as well as other parkinsonian features—for example, tremor and postural instability.^5,6Welders have been reported to be exposed to “a wide variety of potential respiratory hazards”,^7,8 although earlier studies did not show increases in chronic bronchitis.⁹ However, shipyard welders have been reported to have a higher standardised mortality ratio for lung cancer, a finding which was also supported by the California Occupational Mortality Survey, after adjustment for smoking and asbestos exposure. Sjögren presented evidence of a causal relationship between exposure to stainless steel welding and lung cancer.¹⁰Diagnosis of Mn intoxication in workers actively exposed to excessive Mn in welding fumes can be further supported by biomarkers of exposure indicating increased internal Mn concentrations—for example, in whole blood and to a lesser extent in plasma/serum or urine. Sjögren et al¹¹ reported a mean concentration of 8.4 μg/l for whole blood Mn (MnB) in welders, whereas in other industrial settings the mean MnB ranged from 8.1 to 25.3 µg/l. No clear cutoff value of MnB is currently agreed upon, but HAR postulates that any values >10 µg/l is of concern.Mn air levels (Mn‐air) as reported in a few welder studies indicate that aerosols in welding operations (external exposure) usually contain <0.5 mg Mn/m³ (total dust), but sometimes they are >1 mg Mn/m³. A paramount feature of welding fume is that most of the airborne particles are in the respirable fraction (particle size <10 µm).¹² It is astonishing that reliable levels of respirable Mn particulate have rarely been reported, given the importance of the pulmonary uptake of Mn via the alveoli and its significance for Mn distribution to and effects on the central nervous system.¹³ It should be pointed out that inhalation exposure to Mn is for most of the jurisdictions regulated on the basis of total (or inhalable) dust: however, in the particular case of welding, a permissible exposure level established on the respirable fraction would enhance health prevention strategies.Neuropsychological testing methods used over the past two decades have successfully differentiated Mn‐exposed welders from unexposed controls.^4,6,11 There have been 13 reports on welders showing deficits for motor, tremor, memory and neurocognitive domains, as well as for sleep, sexual function and vision. Although not all reports dealt with all of these domains, 11 studies indicated slowing of motor speed/efficiency^{4,6,11,14,15,16,17,18,19,20,21} and tremor^{3,4,6,14,15,16,17,19,20,21,22}; eight found loss of neurocognitive functioning^{4,6,11,14,16,18,19,21}; and six reported diminished memory function.^{4,6,11,14,18,21} Six studies reported sleep disturbances^{4,6,11,14,15,17} and three sexual problems.^4,6,14 Vision was assessed in only two studies, and both found loss of colour vision associated with Mn exposure.^6,14Welders exposed to Mn‐containing welding fumes and dust have rarely been investigated by clinical neuropsychologists using an epidemiological study design in which actual measurements of internal and external exposure to Mn are studied in relation to outcome variables of a comprehensive test battery (including measures of IQ). Therefore, in the latter part of 2004, we decided to evaluate a welder group with health complaints, identified as employees on the reconstruction project of the San Francisco–Oakland Bay Bridge, which was damaged during the 1989 earthquake. The work on a vulnerable portion of the East span of the bridge began in 2003, requiring confined space welding of 28 anchoring foundations or piers, consisting of steel piles and footing boxes. The welding operations took place below the Bay''s surface in unpressurised cofferdams. During the first 1.5 years of welding on the support piers for the new bridge, welders began to express concerns of ill health feared to be the result of exposure to fumes due to confined space welding. During this period the welders were not required to wear personal protective equipment, and ventilation was minimal or ineffective. After receiving health complaints from almost 90% of the welders employed on the project, workers'' compensation evaluations were enacted, which supported their report of adverse health effects. A multidisciplinary study group took advantage of this “natural experiment” for undertaking additional clinical evaluations (1) to assess the extent of neurological, neuropsychological, neurophysiological and pulmonary effects, (2) to explore whether these effects were associated with internal and/or external measures of Mn exposure and (3) to define a tentative permissible exposure level for Mn in welding fume.     

Phase 2 Study of Nimotuzumab in Combination With Concurrent Chemoradiotherapy in Patients With Locally Advanced Non–Small-Cell Lung Cancer

BackgroundWe evaluated the tolerability and efficacy of nimotuzumab, a humanized IgG1 monoclonal anti–epidermal growth factor receptor antibody, with concurrent chemoradiotherapy in patients with unresectable locally advanced non–small-cell lung cancer.Patients and MethodsIn this multicenter, single-arm, open-label, phase 2 trial conducted in Japan (JapicCTI-090825), patients received thoracic radiotherapy (60 Gy, 2 Gy per fraction, 6 weeks) and four 4-week cycles of chemotherapy (day 1, cisplatin 80 mg/m²; days 1 and 8, vinorelbine 20 mg/m²). Nimotuzumab 200 mg was administrated weekly for 16 weeks. The primary endpoint was treatment completion rate, defined as the percentage of patients completing 60 Gy of radiotherapy within 8 weeks, 2 cycles of chemotherapy, and at least 75% of the required nimotuzumab dose during the initial 2-cycle concurrent chemoradiotherapy period.ResultsOf 40 patients enrolled, 39 received the study treatment, which was well tolerated, with a completion rate of 87.2%. Thirty-eight patients completed 60 Gy of radiotherapy within 8 weeks. Infusion reaction, grade 3 or higher rash, grade 3 or higher radiation pneumonitis, or grade 4 or higher nonhematologic toxicity were not observed. The objective response rate was 69.2%. The median progression-free survival (PFS) and 5-year PFS rate were 508 days and 29.0%, respectively. The 5-year PFS rate in patients with non–squamous cell carcinoma (n = 23) was 13.7% and in patients with squamous cell carcinoma (n = 16) was 50.0%. The 5-year overall survival rate was 58.4%.ConclusionAddition of nimotuzumab to the concurrent chemoradiotherapy regimen was well tolerated and showed potential for treating patients with locally advanced non–small-cell lung cancer, particularly squamous cell carcinoma.