bluuebunny/crossref_metadata_2025 huggingface dataset:
Endoscopic treatment of advanced endometriosis with large endometriomata. Surgical technique
Authors: Milingos Spyros , Kallipolitis George , Loutradis Dimitris , Liapi Anthi , Drakakis Peter , Mavrommatis Konstantinos , Antsaklis Aris , Creatsas George , Michalas Stylianos
Published: March 2003

Advanced endometriosis is a common cause of infertility. It has traditionally been treated surgically by laparotomy. The surgeon had to cope mainly with adhesions, endometriotic implants and, quite often, endometriotic cysts of various diameters. In this study, we report our laparoscopic technique for treating advanced endometriosis with large endometriomata, using conventional instrumentation, and we discuss our main problems and our results.

The study population consisted of 68 subfertile patients with large endometriomata who underwent laparoscopic removal of the cysts.

The Infertility Clinic at the Alexandra Maternity Hospital, Athens, Greece, between January 1990 and October 1997.

Laparoscopic excision of endometriotic cysts in subfertile women.

Success of treatment modality; postoperative complications; postoperative cumulative pregnancy rates.

The technique is described. Postoperative complications were minimal. Of the patients, 32 (47%) became pregnant during the 2‐year postoperative follow‐up period. The estimated cumulative pregnancy rates at 6, 12, 18 and 24 months were 22%, 35%, 45% and 47%, respectively.

Our experience favours the use of traditional laparoscopic tools over laser treatment for the management of extensive endometriosis with endometriomata, since the results are similar to those observed after laser treatment, and the training of a larger number of physicians and availability of equipment are more readily achieved for the former method. Postoperative pregnancy rates are high.
Normal saline field bipolar electrosurgery in hysteroscopy: report of the first 163 cases
Authors: Vleugels Michel P. H.
Published: March 2003

To document the clinical and surgical experience of the first large series of hysteroscopic procedures performed with the new bipolar normal saline field electrodes.

Prospective descriptive study.

163 women scheduled for hysteroscopic surgery.

All the scheduled operations could be done with the new device, and 69% of all lesions could be removed under local anaesthesia and 28% without anaesthesia at all. On average, 85 mL of distension fluid, i.e. normal saline, was resorbed. The amount of resorption was related to the operation time, which increased greatly in the case of polyps over 4 cm in size or fibroids larger than 3 cm in diameter. No complications were encountered.

This new bipolar normal saline field device is easy to handle, and safe and effective in removing lesions by coagulation, cutting or vaporization. It can easily be used to perform most hysteroscopic procedures with or without local anaesthesia in daycase settings.
Endoscopically assisted vaginal sacrospinous colpopexy
Authors: Shafik Amina M. , Hefni Mohamed
Published: March 2003

To describe an endoscopic technique which simplifies the transvaginal sacrospinous colpopexy operation for vault prolapse.

Insertion of polydioxanone sutures was carried out under direct endoscopic visualization.

The introduction of this new technique has changed the difficult blind step of the operation into a directly visualized endoscopic procedure and has decreased the likelihood of complications, especially injury to pudendal nerve and vessels and postoperative buttock pain. This could allow the majority of the gynaecologists to safely perform the operation.
Animal toxicities study of a promising new contraceptive device
Authors: Sterling Glenn M.
Published: March 2003

Objective Gore‐tex bags were placed over sheep ovaries and tissue reaction was assessed.

Setting The study was conducted at The University of Queensland under the supervision of the School of Veterinary Science and The Wesley Research Institute.

Design Gore‐tex bags were applied to ovaries at laparotomy and removed after 90 days. Photographs of the gross dissections and histological sections were utilized to document the findings.

Results At repeat laparotomies, there were no adhesions between the ovarian bags and surrounding viscera. The ovaries were viable and surgical planes were easily established between the ovaries, the bags and the outer fibrous capsules. No detrimental tissue reaction was demonstrated in the animal model studied. The bags remained in place for the term of the study.

Conclusion The laparoscopic placement of an inert bag over the ovary may be a method of reliable, reversible, barrier human contraception. The device should be amenable to application and removal using a minimally invasive technique. Other possible applications include oöcyte capture prior to egg collection in assisted reproductive technology and early detection and prevention of ovarian cancer.
Immunoadsorption for removal of inhibitors: update on treatments in Malmö–Lund between 1980 and 1995
Authors: Freiburghaus  , Berntorp  , Ekman  , Gunnarsson  , Kjellberg  , Nilsson
Published: March 2003

Treatment of severe bleeding and the performance of surgery in Haemophilia patients with inhibitors creates severe problems. It is generally agreed that treatment is most effective if circulating levels of factor VIII/IX can be achieved long enough for control of heamostasis. Immunoadsorption with protein A for the removal of inhibitor has improved treatment for patients with initial inhibitor titres too high to neutralize by infusion alone.

 This is a summary of our experience in Malmö regarding immunoadsorption and heamostatis. A total of 19 applications with immunoadsorption in 10 patients were performed. On all occasions it was possible to eliminate totally the inhibitor or reduce it to low levels that could easily be neutralized with factor concentrate. Heamostatic levels of coagulation factors could be maintained for 5–9 days in all but one patient. This period was sufficient to stop ongoing heamorrhage or prevent excessive bleeding at surgical interventions.
Management of a premature infant with moderate haemophilia A using recombinant factor VIII
Authors: Gale  , Hird  , Colvin
Published: March 2003

The birth of a very premature infant with haemophilia A is a rare event. In this case report the problems posed in the management of a child with a factor VIII level of 0.03 IU mL−1 born at 28 weeks of gestation and weighing 1590 g are considered.

The value of recombinant factor VIII, the pharmacokinetics of factor VIII in this situation and the importance of close cooperation between paediatricians and haematologists are discussed.
Hepatic gene therapy for haemophilia B
Authors: Lee C. A. , Kessler C. M. , Varon D. , Martinowitz U. , Heim M. , KAY M. A.
Published: March 2003

Summary. Early retroviral‐mediated factor IX gene transfer into deficient dogs showed that constitutive expression of low levels of factor IX which has led to persistent improvement of clinically relevant parameters such as the WBCT and PTT. Conversely, in vivo adenoviral mediated delivery of the factor IX cDNA into hepatocytes of haemophilia B dogs has resulted in greater than wild‐type plasma concentrations of clotting factor with complete, albeit transient normalization of haemostasis for a short time. An immune response directed against the vector transduced cells presented a big obstacle to clinical application. However, the future of gene therapy for factor IX deficiency appears bright with the development of fully adenoviral‐gene deleted vectors, rAAV and lentiviral vectors which seem to offer safety, therapeutic levels of factor IX and relatively long‐term persistence. We must proceed with cautious optimism as these vector systems undergo further scrutiny.
Safety of coagulation factor concentrates
Authors: Lee C. A. , Kessler C. M. , Varon D. , Martinowitz U. , Heim M. , TEITEL J. M.
Published: March 2003

Summary. Many of the adverse effects of the early crude plasma‐derived concentrates were ameliorated by increasing their purity. Ironically, this strategy may have increased the risks of inhibitor formation and pathogen transmission due to the addition of processing steps which can alter the immunogenicity of clotting factors and the use of very large plasma pools, as dictated by economic considerations. In the absence of extremely sensitive donor screening, these large pools have a high probability of contamination with pathogens, which may be only partially offset by their removal during protein purification. One approach to minimize the risk of viral transmission is to use recombinant clotting factors produced without the use of human or animal plasma proteins at any step in the manufacturing or formulation process. However, as these proteins are synthesized in mammalian cells, even they pose a theoretical risk of pathogen transmission. For plasma‐derived concentrates, the initial viral burden is minimized by screening individual donations and plasma pools with tests which detect virusspecific antibodies, protein antigens, or nucleic acid. These techniques are supplemented by non‐specific viral reduction steps based on physical partitioning and/or inactivation of pathogens which share chemical or physical characteristics. Prion proteins, the putative causative agents of transmissible spongiform encephalopathies, do not share these characteristics with viruses, and it remains to be determined whether they partition into clotting factor concentrates and whether the current strategies can efficiently remove or inactivate them. For all blood‐borne pathogens, active immunization (currently available only for hepatitis B and A) and continued surveillance of susceptible recipients are critical approaches to achieving optimal safety of coagulation factor concentrates.
Continuous infusion therapy in haemophilia
Authors: Lee C. A. , Kessler C. M. , Varon D. , Martinowitz U. , Heim M. , VARON D. , MARTINOWITZ U.
Published: March 2003

Summary. The application of coagulation factor therapy by continuous infusion (CI) was first suggested by Brinkhous in the early 1950s [1]. The recent introduction of this mode of therapy to everyday practice was made possible after the demonstration of a good stability of most factor concentrates which were also found safe regarding potential bacterial contamination. Other developments included a better understanding of the pharmacokinetics of factors concentrates as well as the availability of a new delivery system. Continuous infusion was shown to be superior to bolus injection (BI) in achieving a stabile haemostatic effect, in the prevention of post‐operative bleeding and was found to save between 20–50% in the required factor. This mode of therapy was found effective in haemophilia A and B as well as among patients with inhibitors to FVIII and with von Willebrand disease (vWD).
Resistance to HIV protease inhibitors
Authors: Lee C. A. , Kessler C. M. , Varon D. , Martinowitz U. , Heim M. , CONDRA J. H.
Published: March 2003

Summary. Resistance to the HIV‐1 protease inhibitor indinavir involves the accumulation of multiple amino acid substitutions in the viral protease. A minimum of 11 amino acid positions have been identified as potential contributors to phenotypic resistance. Three or more amino acid substitutions in the protease are required before resistance becomes measurable (≥four‐fold). Further losses in susceptibility follow the stepwise accumulation of additional amino acid substitutions, indicating that antiviral activity (selective pressure) is maintained despite the appearance of multiple amino acid substitutions in the viral protease. Importantly, the sequential nature of these changes indicates that the effects of these substitutions are additive, and that the evolution of resistance is driven by viral replication. This result has significant implications for therapy. It predicts that viral variants resistant to indinavir are unlikely to pre‐exist in protease inhibitor‐naive patients, and further, that high‐level resistance can only develop if the virus is allowed to replicate in the presence of the drug. The use of indinavir in combination with other antiretroviral agents has been demonstrated to dramatically reduce the incidence of resistance mutations, suggesting that with maximal suppression of viral replication, long‐term control of HIV‐1 infection may be achievable. Thus, the goal of therapy must be to never to allow the virus to replicate. This can be best accomplished by initiating therapy with a maximally suppressive regimen, to reduce viral replication as much as possible, and by imposing a high genetic barrier to resistance. Previous use of other protease inhibitors or inadequate adherence to therapy may compromise the long‐term benefit of indinavir by allowing the virus to gain a foothold through the development of resistance. An understanding of these issues will be critical in realizing the full potential of this potent new drug for the control of HIV‐1 infection.
Diagnosis of von Willebrand disease
Authors: Lee C. A. , Kessler C. M. , Varon D. , Martinowitz U. , Heim M. , FEDERICI A. B.
Published: March 2003

Summary. von Willebrand disease (vWD) is a bleeding disorder caused by quantitative or qualitative defects of von Willebrand factor (vWF). vWF is synthesized by endothelial cells and megakaryocytes and circulates in plasma as a multimeric high molecular weight glycoprotein. vWF plays a major role in the early phases of ostasis by promoting platelet‐vessel wall and plateletplatelet interactions under high shear conditions. It is also the carrier of coagulation factor VIII (FVIII) in plasma. A deficiency of vWF results in impairment of both primary and secondary phases of ostasis. Therefore, patients with vWD manifest bleeding symptoms that are typical of defects of primary ostasis (mucocutaneous haemorrhages) but, in case of severe deficiency of vWF, there are also haemarthroses and haematomas, which are typical of those seen with coagulation defects. Several types and subtypes of vWD have been described with a high degree of heterogeneity. The diagnosis is based on measurements of plasma and platelet vWF, the ability of vWF to interact with its platelet receptor and the analysis of the multimeric composition of vWF. Due to the heterogeneity of vWF defects, a correct diagnosis of types and subtypes may be sometimes difficult but is very important for an appropriate treatment of patients with vWD.
Continuous infusion of intermediate‐purity factor VIII in haemophilia A patients undergoing elective surgery
Authors: Rochat  , Mcfadyen  , Schwyzer  , Gillham  , Cruickshank
Published: March 2003

An open, non‐randomized trial of continuous infusion therapy was conducted involving five patients with severe haemophilia A who required factor VIII (FVIII) prophylaxis for elective surgery. This was preceded by a 24‐h preoperative pharmacokinetic study to characterize the pharmacokinetic parameters of each individual patient following a bolus dose of the intermediate‐purity product. A retrospective matched control group was identified to allow for comparisons of FVIII usage between bolus and continuous infusion administration. A loading dose of FVIII was administered preoperatively, and the continuous infusion was started at the end of surgery and continued for 5 days. The patients’ FVIII levels, vital signs, and infusion sites were monitored on a daily basis. The clearance was re‐calculated on a daily basis using the FVIII activity of that day to adjust the infusion rate to achieve the desired FVIII level. The mean (CV%) pharmacokinetic parameters estimated preoperatively by noncompartmental analysis were: clearance 3.2 mL kg−1 h−1 (35.5%), volume of distribution 52.1 mL kg−1 (40.2%), mean residence time 17.4 h (23.3%), and half‐life 12.7 h (23.6%). A progressive decrease in the clearance of FVIII from a mean of 3.1 mL kg−1 h−1 to 2.0 mL kg−1 h−1 (P = 0.125) over the first 5 days was observed. A therapeutically acceptable level of FVIII was systematically achieved, with the only complication being frequent thrombophlebitis. On average the patients used 19% less FVIII when compared with matched historical controls (P = 0.25). This method was found to be safe and effective in haemophilia A patients undergoing elective surgery procedures.
Highly sensitive and species‐specific assay for quantification of human transgene expression levels
Authors: Haack  , Poller  , Schneider‐Rasp  , Thalheimer  , Schmitt  , Hanfland  , Brackmann  , Schwaab
Published: March 2003

During the past few years great efforts have been made to construct and to test human factor VIII (hFVIII) and IX (hFIX) vectors suitable for haemophilia gene therapy in vivo. However, little is known about the molecular mechanisms of persistence and shut‐off of transgene expression in the target organs after gene transfer using recombinant adenoviral vectors. To evaluate low transgene mRNA levels in different tissues, especially at long times after the gene transfer, the common northern blot method is often not sensitive enough. For this reason we developed a new, highly sensitive and species‐specific method for hFIX mRNA quantification and employed it in mice treated with an adenoviral vector (Ad5CMVFIX) expressing human FIX. In addition to its very high sensitivity (lowest detection level=1 fg RNA), the method was shown to be strictly species‐specific, since hFIX mRNA signals were never detected in untreated mice. In a long‐term study of 18 vector‐treated mice we compared the human FIX:Ag levels in the mouse plasma, the human FIX mRNA levels and human FIX vector DNA concentrations in the mouse liver. We found that a slow but continuous decrease of hFIX:Ag levels in mouse plasma was associated with a corresponding decrease of hFIX mRNA levels in the liver. However, the Ad5CMVFIX vector DNA levels did not decrease to a comparable degree, suggesting that the decrease of human FIX:Ag levels in mouse plasma is, to a significant extent, also caused by CMV promotor shut‐off and only to a minor degree by loss of vector DNA.
Gas gangrene in a patient with severe haemophilia A
Authors: Ghosh  , Jijina  , Pathare  , Mohanty
Published: March 2003

The haemophilia patient tends to live a more protected life than his normal counterpart, this is particularly so in underdeveloped and developing countries where due to poor health infrastructure, financial constraints and nonavailability of factor concentrates, patients quickly learn that they need to live a protected life. Under such circumstances, gas gangrene seems to be a very unusual infection for this group of patients. We report here a 25‐year‐old male with severe haemophilia who developed gas gangrene due to inadequate medical management following a road traffic accident. Subsequently, his affected limb was salvaged by conservative therapy. A literature search failed to reveal any reports of similar patients in the English literature.
Major surgery for a gastric cancer in a haemophilic with high inhibitor titre successfully performed by the use of recombinant FVIIa
Authors: Rocino  , Carola  , Papa  , Parisi  , Cremona  , Miraglia  , De Biasi
Published: March 2003

A total gastrectomy with omentectomy and resection of the distal oesophagus in a 69‐year‐old haemophilia A patient with high inhibitor of 128 Bethesda units is described. Surgery was successfully performed after infusion of 112 μg kg–1 bw of recombinant FVIIa. Ninety‐two μg kg–1 were given thereafter at time intervals of 2 h until 12 h, then every 3 h until 24 h, and every 4 h until 48 h after surgery. Doses were gradually reduced in the following days and finally discontinued on day 28 after surgery. The complete treatment schedule required the administration of a total of 708 mg of recombinant FVIIa. Using this approach, we observed normal haemostasis, and there were no signs of excessive postoperative bleeding or wound haematoma. No clinical side‐effects or evidence of systemic activation of coagulation occurred during the treatment. As judged from the clinical course of this major surgery, recombinant FVIIa appears to be highly efficacious and safe and should be used as first line treatment in high titre inhibitor patients with cross‐reactivity to porcine factor VIII, undergoing surgery.
Therapeutic options in the management of articular contractures in haemophiliacs
Authors: Rodriguez‐Merchan E.C.
Published: March 2003

Haemophilic contracture is seen most commonly as an equinus deformity of the ankle, or at the knee or elbow in the form of a flexion deformity. Treatment options are varied, and decision‐making is based on the degree of the contracture, its chronicity, the presence of articular subluxation, the patient’s ability to participate in treatment, and the available medical facilities. The treatments available fall into four categories: physiotherapy, orthotics, corrective devices, and surgical procedures. Treatment should be primarily by physiotherapy, splintage, and corrective devices. The late or severe case may require surgical correction in the form of soft‐tissue procedures. Soft‐tissue correction of muscle shorthening may be performed such as lengthening of the Achilles tendon for equinus deformity of the ankle, or hamstring release of the flexor muscles of the knee. Lower femoral osteotomy has been used for correction of flexion deformity at the knee joint. Mechanical distraction using external fixators for treatment of severe knee flexion contractures has been recently reported with satisfactory results. The main principle underlying the treatment of haemophilic contracture is the restoration of the patient’s lifestyle and mobility, rather than anatomic or radiographic normality.
Management of fixed flexion contracture of the elbow in haemophilia
Authors: Gilbert M.S. , Radomisli T.E.
Published: March 2003

The authors stress that prevention of flexion contractures and artropathy by early factor replacement and physical therapy for every haemophiliac is the standard of care. Physical therapy, serial casting, and Quengel cast correction have not proven successful in correction of fixed flexion contractures at the elbow. In the patient who has a flexion contracture that interfered with function, an attempt at physical therapy combined with the use of either the Dynasplint or Flowtron will be tried. If there is no response, a surgical synovectomy combined with a possible radial head resection and anterior capsular relase would be the authors’ procedure of choice. In the face of advanced arthropathy, the authors would consider a distraction arthroplasty.
von Willebrand factor containing factor VIII concentrates
Authors: Berntorp E.
Published: March 2003

There are several plasma derived von Wille‐brand factors (vWF) containing factor (FVIII) concentrates that can be used in the treatment of von Willebrand disease (vWD). All concentrates are effective in attaining normal postinfusion levels or of FVIII:C but it is difficult to achieve normalization of the bleeding time even with concentrates containing almost all vWF multimers including those of high molecular weight. Haemate P (Centeon) may be considered as the golden standard concentrate available at present. However, the development of more purified vWF concentrates devoid of FVIII:C is the goal for future development.
Making a therapeutic choice: human versus recombinant fractions – can we do it?
Authors: Aledort L.M.
Published: March 2003

New manufacturing methods have resulted in production of higher purity human‐derived products and recombinant factors with less human‐ or animal‐derived reagents or stabilizers for the treatment of congenital coagulation disorders. In addition, new viral inactivation processes are being used to reduce the chance of viral contamination. Preferred treatment approaches are moving toward using newer‐generation agents. At the same time, rationale exists for continuing fractionation of all types of products, based on considerations including cost, product availability, safety issues, and specific clinical situations for which a recombinant or high‐purity product is not available. These issues and others are reviewed in the context of making a therapeutic choice.
Early transfusion of factor VIII/von Willebrand factor concentrates seems to be effective in the treatment of gatrointestinal bleeding in patients with von Willebrand type III disease
Authors: Zanon E. , Vianello F. , Casonato A. , Girolami A.
Published: March 2003

The association between gastrointestinal angiodysplasia and von Willebrand disease was reported 30 years ago. The clinical course of patients with von Willebrand disease and angiodysplasia is characterized by numerous admissions to hospital for gastrointestinal bleeding necessitating transfusion with packed red cells, factor VIII and plasma. The management of these patients is problematic. Numerous treatments for the gastrointestinal bleeding have been proposed: surgery, electrocoagulation, laser photocoagulation, sclerotherapy, arteriography with embolization, immunoglobulins, oestrogens, and octreotide, but no treatment modality has been successful in all cases. We report a 66‐year‐old‐female with small bowel angiodysplasia and von Willebrand type III disease in whom prompt administration of factor VIII/vWF concentrates was effective. Education of patients to recognize minimal gastrointestinal bleeding manifestations, periodical clinical visits and early infusion of factor VIII/vWF seems to be fundamental for the success of this therapy. A longer follow‐up and the study of other patients are needed to confirm our observation.
Changes in treatment strategies for severe haemophilia over the last 3 decades: effects on clotting factor consumption and arthropathy
Authors: Fischer K. , Van Der Bom J. G. , Mauser‐Bunschoten E. P. , Roosendaal G. , Prejs R. , Grobbee D. E. , Van Den Berg H. M.
Published: March 2003

A cohort study was performed among 214 patients with severe haemophilia, born 1944–1994, to describe changes in treatment over the last 3 decades and its effects on clotting factor consumption and haemophilic arthropathy. Data on treatment strategy, clotting factor consumption, and outcome were collected for 3567 patient years (from 1972 to 1998), and 493 Pettersson scores were analysed. Median follow up was 17 years (range 6–27 years), and median age in 1998 was 27.6 years. Since 1965, replacement therapy, prophylaxis, and home treatment have been used and treatment intensified. Over the last 3 decades, annual clotting factor consumption increased by 260%, for both prophylactic and on‐demand treatment. Annual clotting factor consumption kg–1 increased during childhood and appeared to stabilize in early adulthood for patients born 1965–79, who were treated with early replacement therapy or early prophylaxis. In contrast, clotting factor consumption increased continuously for patients born before 1965, who had had no access to replacement therapy during the early years of their life. The annual number of joint bleeds decreased over the years. Arthropathy as measured by the Pettersson score generally became apparent around the age of 15 years and was lowest in patients treated with primary prophylaxis. In conclusion, clotting factor consumption has increased and haemophilic arthropathy has decreased due to the intensification of treatment for severe haemophilia over the last 3 decades. Annual clotting factor consumption stabilizes in adulthood for patients who receive early intensive treatment.
Ophthalmic surgery in haemophilia
Authors: Jijina F. , Ghosh K. , Madkaikar M. , Mohanty D.
Published: March 2003

A wide range of ophthalmic surgical procedures were conducted in five patients with haemophilia of varying severity (one severe and four mild) aged between 8 and 75 years. The operations included intraocular lens implantation, trabeculectomy and vitrectomy. Successful postoperative outcome with haemostasis was achieved in all patients with moderate use of clotting factor concentrates (3000–7000 IU), simultaneous use of oral epsilon amino caproic acid therapy and intravenous deamino‐8‐D‐arginine vasopressin (desmopressin; DDAVP) wherever feasible. None of the patients had circulating inhibitor. One of the patients with milder disease (FVIII 32%) was referred to us after he was operated on for hyphaema elsewhere, without prior knowledge of his diagnosis of haemophilia. Thus, satisfactory eye surgery in patients with haemophilia is possible with a restricted amount of factor concentrates with gratifying results.
Effects of HAART on hepatitis C, hepatitis G, and TT virus in multiply coinfected HIV‐positive patients with haemophilia
Authors: Takamatsu J. , Toyoda H. , Fukuda Y. , Nakano I. , Yokozaki S. , Hayashi K. , Saito H.
Published: March 2003

In multiply coinfected human immunodeficiency virus (HIV)‐positive patients, we investigated the effects of high‐activity antiretroviral therapy (HAART) using HIV protease inhibitors on three other viruses: hepatitis C virus (HCV), hepatitis G virus (HGV), and TT virus (TTV). Viral concentrations were measured serially by polymerase chain reaction methods in five patients with quadruple infection (HIV, HCV, HGV, and TTV) and in two patients with triple infection (HIV, HCV, and HGV) before and during HAART. In addition, CD4+ cell counts and serum alanine aminotransferase (ALT) levels were measured serially. Generally we observed no difference in serum HCV RNA, HGV RNA, or TTV DNA concentrations between samples obtained before and after initiation of HAART, whereas HIV RNA concentration decreased and CD4 counts increased in most patients. However, two patients had markedly decreased concentrations of HCV RNA and HGV RNA, respectively, more than 12 months after beginning HAART. Normalization of serum ALT levels was observed in a patient with decline of HCV RNA concentrations. No interactions were observed among these four viruses. HAART had no apparent direct effects on HCV, HGV, or TTV. Further studies will be required to elucidate whether the restoration of immune status through suppression of HIV replication by HAART may affect HCV or HGV RNA concentrations.
Development of gene therapy for blood disorders by gene transfer into haematopoietic stem cells
Authors: KARLSSON S. , OOKA A. , WOODS N‐B.
Published: March 2003

Haematopoietic stem cells (HSCs) are important target cells for gene therapy of blood disorders due to their pluripotency and ability to reconstitute haematopoiesis following myeloablation and transplantation. HSCs can `self‐renew' and generate new stem cells. Genetically modified stem cells are therefore expected to last a lifetime in the recipient following blood and marrow transplantation, and can potentially cure haematological disorders. Oncoretroviral vectors have been the main vectors used for HSCs because of their ability to integrate into the chromosomes of their target cells. Because oncoretroviral vectors require dividing target cells for successful localization of the preintegration complex and subsequent chromosomal integration of the provirus, only the dividing fraction of the target cells can be transduced. As only a small fraction of haematopoietic stem cells is dividing at any one time, oncoretroviral vector transduction of human HSCs has been low in clinical trials. However, patients with severe combined immune deficiency‐X1 (SCID‐X1) have recently been treated successfully by gene therapy of autologous bone marrow cells using oncoretroviral vectors containing the common γ chain gene. While several additional disorders may potentially be treated successfully using oncoretroviral gene transfer to HSCs, many disorders may require much higher gene transfer efficiency than was achieved in the SCID‐X1 study. Therefore, lentiviral vectors have recently emerged as promising vectors for human HSCs because they can transduce dividing and nondividing HSCs efficiently, and may become the vectors of choice in the future for treatment of blood disorders where a large fraction of HSCs has to be corrected.
Carrier detection and prenatal diagnosis in haemophilia in India: realities and challenges
Authors: GHOSH K. , SHETTY S. , PAWAR A. , MOHANTY D.
Published: March 2003

Organizing services for haemophilia in developing countries with few resources is a formidable task. There is wide variation in haemophilia care and management between developing and developed countries. The management of a genetic disorder such as haemophilia becomes difficult in developing countries where scanty resources are allocated mainly to nutrition and infectious diseases as a first priority. In a country such as India, with one billion people and with a wide diversity in cultural, educational and financial conditions, educating people about such diseases is difficult and will take a long time to have an effect on attitudes. Meanwhile, attempts are being made at several centres in India to develop carrier detection and prenatal diagnosis for this disease. Counselling a carrier for detection of her carrier status remains problematic in India because marriages are still largely arranged by the parents of prospective brides and bridegrooms. Hence, the very idea of communicating the carrier status to the bridegrooms' families may lead to cancellation of the marriages, and concealing the carrier status is associated with guilt both for the bride's parents as well as the bride. Prenatal diagnosis and carrier detection in India therefore must be discussed in this context.
Advances in the therapy of von Willebrand disease
Authors: BATLLE J. , NOYA M. S. , GIANGRANDE P. , LÓPEZ‐FERNÁNDEZ M. F.
Published: March 2003

von Willebrand disease (vWD) is a very common autosomal inherited bleeding disorder, caused by a quantitative deficiency or a qualitative structural defect of von Willebrand factor (vWF). Two main therapeutic options are available for the treatment of spontaneous bleeding episodes and for prevention of bleeding: desmopressin (DDAVP) and replacement therapy with plasma products. DDAVP is the treatment of choice for most patients with type 1 vWD. In patients with the type 3 disease and in most subjects with type 2 disease, DDAVP alone is ineffective or contraindicated, and it is usually necessary to switch to plasma concentrates containing both factor VIII (FVIII) and vWF. Concentrates subjected to virucidal treatment (e.g. solvent/detergent treatment) during manufacture should always be used in preference to cryoprecipitate. A recombinant vWF concentrate is now undergoing preclinical development and preliminary data suggest it possesses good haemostatic function and may correct the bleeding in vWD after its administration in several animal models. Although treatment of vWD is relatively simple (assuming access to even basic laboratory facilities), actual diagnosis is often far from straightforward, and the patients should be well characterized phenotypically to tailor the treatment to the different types and subtypes of the disease. It is probably wise to refer samples for the characterization to expert laboratories.
Type 2B vWD: the varied clinical manifestations in two kindreds
Authors: Mathew P. , Greist A. , Maahs J. A. , Lichtenberg E. C. , Shapiro A. D.
Published: March 2003

Type 2B von Willebrand's disease (vWD) is associated with spontaneous binding of large von Willebrand factor (vWF) multimers to platelets in vivo, followed by clearance of both the large multimers and platelets resulting in thrombocytopenia, which may be intermittent, mild to severe, and may be exacerbated by stress such as infection or pregnancy. We report our experience in two kindreds (49 caucasian individuals) with type 2B vWD and discuss their varied clinical manifestations. The largest kindred (45 patients) was traced back five generations to a presumed index case. The genetic defect in this kindred was identified as a missense mutation, with a C to T transition at a CpG dinucleotide (nucleotide 3916) resulting in an amino acid substitution (Arg 543 to Trp) within the glycoprotein Ib binding domain of vWF. Ristocetin cofactor activity varied from < 10 to 28%, and factor VIII activity from 7 to 69%. Analysis of von Willebrand multimers consistently revealed loss of large molecular weight forms. Platelet counts in those with thrombocytopenia varied from 10 × 109 L−1 to 120 × 109 L−1. The severity of thrombocytopenia has also varied within the same individual during the period of follow‐up. The clinical manifestations were varied and ranged from mild to moderate spontaneous bleeding episodes, including epistaxis, menorrhagia and gastro‐intestinal haemorrhage. Severe bleeding episodes were observed in those undergoing surgery (both elective and non‐elective), and in a few patients despite aggressive replacement with exogenous source of intact vWF, antifibrinolytics when indicated, and a near normal platelet count. Thrombotic disease may be a rare and unusual sequela of this disorder as was noted in one of our patients. Obtaining a platelet count at birth in infants of mothers with type 2B vWD who exhibit thrombocytopenia, may help in the earlier detection of infants at risk for thrombocytopenia.
Applying natural forestry concepts in an intensively managed landscape
Authors: Peterken G. F.
Published: March 2003

Near‐to‐nature forestry is an attractive concept, but it is difficult to apply in a landscape where (i) the remaining fragments of original‐natural forests are small and far apart; (ii) most large forests are secondary and very artificial; (iii) long‐naturalized trees and other forest species are present throughout the country, and; (iv) natural disturbance regimes and their consequences cannot readily be observed. This paper summarizes the issues and problems relating to ancient seminatural woods and conifer plantation forests in Britain. Measures are indicated that would bring their management nearer to nature. Natural elements can be incorporated into stand composition, stand structure, processes and patterns, provided general principles are interpreted in a pragmatic and flexible manner.
Targeting squares for survey: predicting species richness and incidence of species for a butterfly atlas
Authors: Dennis Roger L. H. , Hardy Peter B.
Published: March 2003

1.Predictions on species richness and incidence of species are made using data for three scales of mapping from the Greater Manchester butterfly atlas; for the whole of the conurbation (2 km×2 km scale) and for two sample areas centred on the Mersey Valley (1 km×1 km scale; 1 ha scale). Predictions are based on data for recording effort, altitude, biotopes, host‐plants and nectar resources.

2.Data for Greater Manchester indicate that substantial shortfalls may occur in recording butterfly species for atlases despite the fact that butterflies are generally easily identified and well supported with recorders. Shortfalls tend to be larger for species with fewer records, indicating that some species may be more easily overlooked than others.

3.The results demonstrate that targeting squares for re‐survey is necessary and feasible. The predictions have other valuable research applications, the most important of which is being able to assess the accuracy of distribution maps, to correct them, and to make projections of distribution changes.

4.Predictions may be enhanced by improvements to mapping in three ways: (i) Collecting data on recording effort. Variation in recording effort typically accounts for differences in species richness and incidence of species more than any other variable; (ii) Collecting data on biotopes and specific resources. The present results are promising and demonstrate that the collection of environmental data linked to a suitable sampling frame could facilitate knowledge of the distribution of species over extensive areas that remain under‐recorded; and (iii) Distinguishing between breeding individuals and vagrants. Vagrancy is a problem associated both with species and scale. Although species vary substantially in their capacity to migrate beyond their habitats, the effect of vagrancy on distribution maps becomes an increasingly large problem as the grain of mapping (size of recording units) decreases. It is suggested that over‐recording can be a problem, particularly when mapping is fine‐grained.
Threshold levels of purified natural Bos d 2 for inducing bronchial airway response in asthmatic patients
Authors: Zeiler T. , Taivainen A. , Mäntyjärvi R. , Tukiainen H. , Rautiainen J. , Rytkönen‐Nissinen M. , Virtanen T.
Published: March 2003

Background Provocation tests are invaluable in establishing threshold levels and a causal relationship between atopic asthma and a certain allergen source, especially in relation to work‐associated exposure. Purified major allergens open possibilities for a more accurate assessment of sensitization.

Objective To determine the threshold dose of purified major bovine dander allergen Bos d 2 in bronchial provocation in comparison with the standard allergen and a set of other parameters of allergy.

Method Nine consecutive patients referred to hospital for confirming the bovine origin of their occupational asthma were subjected to bronchial provocation tests with purified natural Bos d 2 and a standard bovine dander allergen. Additional tests included bronchial histamine challenge, measurements of total IgE, specific IgE antibody determinations and skin prick tests (SPT) with both allergens.

Results In the provocation tests with Bos d 2, a 15% decrease in the forced expiratory volume in 1 s (FEV1) and/or peak expiratory flow (PEF) values in eight out of nine patients confirmed the predominant role of Bos d 2 in the sensitization. The threshold dose of Bos d 2 varied from 0.1 µg to > 100 µg (median ± median absolute deviation = 4.5 ± 3.9 µg). A positive SPT was induced by a median dose of 13.9 ± 9.8 µg of Bos d 2. Bronchial response to histamine and IgE antibodies against Bos d 2 showed the highest correlations to the provocations results.

Conclusions The efficacy of Bos d 2 in bronchial provocation in patients with occupational cattle‐associated asthma was confirmed and the range of the threshold level was determined. There were individual variations, but the response in provocation remains the reference method for identification of the cause of occupational atopic asthma. SPT and the measurement of specific IgE antibodies, preferably with purified or recombinant major allergens, increase the accuracy of the diagnosis.
Genetic variation of Der p 2 allergens: effects on T cell responses and immunoglobulin E binding
Authors: Hales B. J. , Hazell L. A. , Smith W. , Thomas W. R.
Published: March 2003

Background Der p 2 is a highly polymorphic allergen that shows a distinct pattern of sequence divergence. The effect of the variations on T cell and antibody responses has not been compared.

Objectives To compare IgE antibody binding and T cell proliferation and cytokine release induced by variants of Der p 2.

Methods Peripheral blood mononuclear cells (PBMC) from 19 allergic and 15 non‐allergic people were stimulated with recombinant variants of Der p 2. IL‐5, IL‐10, IL‐13 and IFN‐γ were measured by a time resolved fluorescence (TRF) assay. Serum IgE antibody was measured using a solid‐phase TRF assay.

Results Overall the most prevalent variant of Der p 2 (Der p 2. 0101) was the highest or approximately equal highest inducer of T cell proliferation and IL‐5, IL‐10, IL‐13 and IFN‐γ release. The most divergent variant 0104 induced the next highest responses. The variants 0107 and 0108 showed interesting changes especially when the allergic status was considered. Responses to 0107 showed poor Th1/Th2 polarization and, except for IL‐10 release, cytokine responses to 0108 were low for non‐allergic subjects. The variant 0101 showed similar monoclonal antibody binding but moderately less IgE binding than the other variants.

Conclusions The most prevalent variant, Der p 2. 0101, was the most active for T cell stimulation and although its IgE binding was slightly less than other variants that was highly correlated. The variant Der p 2. 0104 which contains the known common polymorphic changes had a response which was similar to Der p 2. 0101 and thus these two variants were the most stimulatory respresentations of Der p 2. The T cell responses to the less common variants 0107 and 0108 however, showed consistent differences demonstrating that changes in the sequence could change the cytokine response.
Lower airway inflammatory responses to repeated very‐low‐dose allergen challenge in allergic rhinitis and asthma
Authors: Boulay M.‐E. , Boulet L.‐P.
Published: March 2003

Background Low‐dose allergen challenge (LDAC) may be a useful tool for studying the capacity of allergens to induce airway inflammation in atopic subjects.

Objective To evaluate lower airway inflammatory changes following repeated inhalation of very low doses of allergen (VLDAC) in non‐asthmatic subjects with allergic rhinitis (NAAR) compared with mild allergic asthmatic subjects (AA).

Methods Fourteen NAAR and 11 AA were seen out of the pollen season and had skin prick tests with common aeroallergens. Baseline spirometry (S) and methacholine challenge (MC) were done and blood and induced sputum (IS) differential cell counts were obtained. Each subject underwent VLDAC on four consecutive mornings with a relevant allergen. S, MC, and blood and IS samplings were repeated 6 h after the second and fourth VLDAC and one week later.

Results Although there were, as expected, no changes in FEV1 or PC20 in either group, mean percentage eosinophils on IS were significantly increased in NAAR on day 2 of VLDAC and decreased in all but one subject on day 4, with a tendency to return to baseline levels one week later. In AA, there was a non‐significant trend for sputum eosinophils to increase on day 2; four subjects showed a decrease of eosinophils on day 4 of VLDAC. There was a correlation between eosinophil cationic protein (ECP) levels and eosinophil counts in NAAR throughout the study. There were no variations in other sputum cells or blood inflammatory cells.

Conclusion VLDAC can increase the percentage of eosinophils in IS of NAAR subjects without associated respiratory symptoms nor physiological modifications. A reduction in eosinophilic response despite repeated exposure, more common in NAAR subjects, suggests an adaptation process that needs to be further evaluated.
Size‐related deterioration of semi‐natural grassland fragments in Sweden
Authors: Kiviniemi Katariina , Eriksson Ove
Published: March 2003

Abstract.One of the most dramatic landscape changes during the 20th century in Sweden, like in most of Europe, has been the reduction and fragmentation of semi‐natural grasslands. Using a set of remnant semi‐natural grasslands, chosen to be as similar as possible, but differing in size, we have examined whether size of remnant fragments of traditionally managed semi‐natural grasslands in Sweden is related to patterns of species richness and composition. We focused on edge‐to‐interior relationships, since we expected that a possible impact from invasive habitat generalists would be manifested in a gradient from the edge of fragments to their interior. We found no relationship between size of grassland fragments and (a) overall species richness, (b) species richness at different spatial scales, and (c) abundance of some typical invader species or species characteristic of semi‐natural grasslands. However, the results indicated that larger grasslands have a comparatively larger number of species in the edges, whereas the opposite pattern was found in smaller grasslands. The similarity in species composition between the edge and the interior of the pastures also increased with grassland size. Thus, even though the overall species richness is still unaffected by reduction in grassland fragment size, the edges of smaller grasslands show signs of degradation, i.e. reduction in species richness and a decreased similarity to the grassland interior. We suggest that these kinds of effects may be early signs of fragmentation effects that in the future will result in species loss even if the present distribution of semi‐natural grasslands is maintained.