Inhibition by mitoxantrone of in vitro migration of immunocompetent cells: a possible mechanism for therapeutic efficacy in the treatment of multiple sclerosis

BACKGROUND: Damage of the blood-brain barrier and invasion of immunocompetent cells into the central nervous system represent key events in the immunopathogenesis of multiple sclerosis. Mitoxantrone hydrochloride reduces progression of disability and clinical exacerbations in patients with multiple sclerosis. Its precise mode of action is unclear. OBJECTIVE: To investigate the effects of mitoxantrone on the migratory capacity of immunocompetent cells ex vivo and in vitro. DESIGN: Case-control study. SETTING: Department of Neurology, Heinrich Heine University, Düsseldorf, Germany. PARTICIPANTS: Peripheral blood mononuclear cells (PBMCs) were obtained from 11 patients with multiple sclerosis before and after intravenous mitoxantrone treatment; PBMCs from 5 healthy control donors were treated with mitoxantrone in vitro. MAIN OUTCOME MEASURES: The migratory capacity was studied in an in vitro Boyden chamber assay; cells and their rates of migration were analyzed by light microscopy and flow cytometry. To determine the specificity of our findings, PBMCs were treated with perfosfamide in vitro. RESULTS: Mitoxantrone decreased the migratory capacity of CD14(+) monocytes and (to a lesser degree) of CD4(+) and CD8(+) T lymphocytes. These observations were confirmed when control PBMCs were treated with an equivalent dose of mitoxantrone in vitro. Similar effects were seen when PBMCs were preincubated with perfosfamide. The inhibitory effects of mitoxantrone on the migratory capacity of PBMCs were mediated by reduced matrix metalloproteinase 9 activity, as demonstrated by zymography, polymerase chain reaction, and inhibitory studies. CONCLUSION: Mitoxantrone may inhibit the migration of inflammatory cells into and within the central nervous system.     

Cerebral hemodynamics in neurocirculatory hypotension]

Data are presented on the state of cerebral haemodynamics in patients with neuro-circulatory hypotension that was studied by measuring the arterial pressure in various zones of blood supply (the central retinal artery, superficial temporal artery and the brachial artery), by studying the ocular fundus and by rheoencephalography. The cerebral haemodynamic disorders were revealed in the form of regional cerebral hypertension in 13.4% and regional cerebral hypotension in 29.4%, against the background of a general fall of the arterial pressure. In regional cerebral hypertension the vessels of the ocular fundus were narrowed, in regional cerebral hypotension--vice versa. The reduction of the pulse pressure during neuro-circulatory hypotension also attracts attention, and it can be considered a sign of impaired blood supply to the brain.     

War Injuries of Major Extremity Arteries

Abstract During the period 1991–1994, 99 patients (all males, median age 35 years) with combat-related injuries of major limb arteries were managed. Mechanism: mine fragments (40%), high-velocity projectiles (35%), and shotgun pellets (25%). Patients were admitted 1 hour to 16 hours (median 8 hours) after injury; 39% were in severe hemorrhagic shock. Arterial injury was diagnosed by clinical findings. Preoperative angiography was usually not necessary. Of 99 injured patients, 50 (51%) showed signs of distal ischemia and required arterial reconstruction. No primary amputation was performed. Reconstruction was always necessary in cases of injury of axillary or popliteal arteries, but not of superficial femoral or brachial arteries. Ligation of injured single forearm or crural arteries was well tolerated. End-to-end anastomosis by reconstruction was possible only in 38% of cases. In 56% of patients, autologous venous bypass was performed. Uncontrolled wound infection developed in 22% of cases, leading to a secondary hemorrhage compelling arterial ligature (8%), and thrombosis (6%). The secondary amputation rate after arterial reconstruction was 10%. Injury of major vessels was associated with limb bone fractures, nerve damage, or major vein injuries in 68% of cases, frequently on the forearm, the popliteal region, and the crural region. When limb ischemia was present, vascular reconstruction had priority over orthopedic immobilization. Arterial injury was almost always associated with the venous damage in the forearm, the popliteal region, and the crural region. Injured veins of the upper limb were ligated; venous repair was more often indicated in lower limb injury (52%). The method of choice was lateral suture/patch. Gunshot damage to peripheral nerves was rarely treated with primary repair.     

Cladribine impedes in vitro migration of mononuclear cells: a possible implication for treating multiple sclerosis

Background:  Damage of the blood–brain barrier and the migration of immunocompetent cells into the CNS represent key events in the immunopathogenesis of multiple sclerosis (MS). Cladribine is an immunosuppressive drug currently investigated in a phase-III clinical trial for relapsing–remitting MS. However, its precise mode of action remains elusive so far.
Methods:  Peripheral blood mononuclear cells (PBMCs) were isolated from five patients with MS and five healthy donors. PBMCs were treated with cladribine in vitro . The migratory capacity was studied in an in vitro Boyden chamber assay; cells and their rate of migration were analyzed by light microscopy and flow cytometry.
Results:  Cladribine decreased the migratory capacity of CD14⁺ monocytes, as well as of CD4⁺ and CD8⁺ T lymphocytes. T lymphocytes were affected more than monocytes. There was no difference in this effect when comparing mononuclear cells from MS patients with cells from healthy controls.
Conclusions:  Cladribine might achieve, at least in part, its clinical and paraclinical efficacy by inhibiting the migration of inflammatory cells into and within the CNS.     

Neuroendocrine pancreatic tumor in a patient with dual diagnosis of tuberous sclerosis complex and basement membrane disease: A case report and review of the literature

Tuberous Sclerosis is a complex genetic disease that has well-defined clinical criteria. These criteria don''t include pancreatic neuroendocrine tumors. We represent a rare case of a patient, with a non–functioning pancreatic neuroendocrine tumor and concomitant diagnosis of tuberous sclerosis complex, and basement membrane disease.The patient was diagnosed based on typical radiologic findings. We have suggested close monitoring and during follow-up studies, the disease was stable. Interestingly the patient tested negative for Tuberous Sclerosis Complex (TSC), which suggests that she might be a somatic mosaic and the mutation level in blood lymphocytes was below the detection level. Moreover, a heterozygous pathogenic variant p.(Gly774Arg) and a heterozygous likely pathogenic variant p.(Gly1465Asp) were identified in the COL4A4 gene. COL4A4 gene is responsible for causing autosomal dominant basement membrane disease. In this case report, we discuss clinical, radiologic, and genetic aspects of these diseases, as well as optimal treatment and follow-up strategies. Thus, by presenting this case we would like to increase awareness of pancreatic neuroendocrine tumors in TSC and emphasize the need for follow-up monitoring.