Expression of hOvol2 in the XY body of human spermatocytes.

Male infertility is common at infertile clinics, and 10-20% of infertile males are azoospermic. Non-obstructive azoospermia is a complex multifactorial disease, and the process of spermatogenesis remains largely unknown. Ovol1 and Ovol2, a family of zinc finger transcription factors, are expressed in spermatocytes at the pachytene stage and are suggested to be critical regulators of pachytene progression in male germ cells. In this study, we examined the expression of human Ovol2 (hOvol2) in the seminiferous tubes of patients subjected to testicular sperm extraction. We first cloned hOvol1 and hOvol2 from the testis of one of the patients and found no alteration in these nucleotide sequences of this patient. While hOvol1 and hOvol2 were detected by RT-PCR in the testis of patients capable of spermatogenesis, they were not detected in those with Sertoli cell-only syndrome. We recently succeeded in preparing anti-Ovol2 antibody by immunising rats with recombinant mouse Ovol2 (mOvol2) and confirmed the specificity and cross-reactivity of this antibody with hOvol2 in cells transfected with hOvol1 or hOvol2 cDNA. hOvol2 expression was restricted to the XY body of spermatocytes at the pachytene stage. This study demonstrates that hOvol2 is expressed in germ cells and may be involved in spermatogenesis.

Characteristic endoscopic findings of gastrointestinal malignant lymphomas other than mucosa-associated lymphoid tissue lymphoma.

Background and study aims: The gastrointestinal (GI) tract is the most common site of extra-nodal involvement for non-Hodgkin's lymphoma (NHL). The features of GI NHLs remain unclear. The aim of this study was to clarify endoscopic characteristics of GI NHLs. Patients and methods: We retrospectively analyzed the morphological characteristics of 63 GI malignant lymphomas other than mucosa-associated lymphoid tissue lymphoma. Lesions were diagnosed between 2005 and 2020. Macroscopic findings were classified into five subtypes: superficial (S); protruding without ulcer (P); protruding with ulcer (PU); fungating (F); and multiple nodules (MN). Results: Thirty-one lesions in the stomach were classified as S type in 3 cases (9.6%), P type in 6 (19%), PU type in 13 (42%), and F type in 9 (29%). In the stomach, the ulcerated phenotype was more frequent for diffuse large B-cell lymphoma (DLBCL) (89.5%) than for other histological types (41.7%; P = 0.01). In the intestine, 23 tumors were classified as S type in 4 cases (17%), P type in 1 (4%), PU type in 6 (26%), F type in 1 (4%), and MN in 11 (48%). Eleven of the 14 cases (78.6%) of intestinal follicular lymphoma lesions showed MN type. In the colon, eight tumors were classified as S type in 2 cases (25%), P type in 2 (25%), PU type in 1 (13%), and F type in 3 (38%). Conclusion: We have clarified the endoscopic features of GI NHL using macroscopic classifications. The ulcerated phenotype was the most frequent endoscopic finding for DLBCL.

Genetic diversity and population structure of Indonesian native chickens based on single nucleotide polymorphism markers.

Indonesian native chickens are considered an important genetic resource, particularly with respect to their excellent traits for meat and egg production. However, few molecular genetic studies of these native chickens have been conducted. We analyzed the genetic diversity and differentiation of 4 populations of Indonesian native chickens: Black Kedu (BK), Kedu (KD), Kampung (LOC), and Arab (AR). Blood samples from 188 individuals were collected in central and western Java. Genomic DNA was genotyped using 98 autosomal SNP markers, of which 87 were found to be polymorphic. The proportion of polymorphic loci and the average heterozygosity of each population were in the range of 0.765 to 0.878 and 0.224 to 0.263, respectively. The 4 populations of Indonesian chickens appeared to be derived from 3 genetic populations (K = 3): maximum likelihood clustering showed that the BK variety and AR breed were each assigned to a distinct cluster, whereas the LOC ecotype and KD variety were admixed populations with similar proportions of membership. Principal components analysis revealed that eigenvector 1 separated BK and AR from the other 2 populations. Neighbor-joining trees constructed from pairwise distance matrix (F(ST)) estimates, for individuals and between populations, corroborated that the LOC ecotype and KD variety were related closely, whereas the BK variety and AR breed diverged at greater distances. These results also confirmed the usefulness of SNP markers for the study of genetic diversity.

Involvement of stem cell factor and its receptor tyrosine kinase c-kit in pain regulation.

The c-kit receptor tyrosine kinase is expressed in a subpopulation of small- and medium-sized neurons of the dorsal root ganglia (DRG) and in the superficial layer of the spinal cord. Stem cell factor (SCF), a ligand of the c-kit receptor, induces neurite outgrowth from DRG and supports the survival of c-kit-expressing neurons. To clarify the possible function of the SCF/c-kit receptor system in the adult animal, we investigated the expression of c-kit receptor in the spinal cord and DRG in relation to pain by using H2C7, a newly developed anti-c-kit monoclonal antibody. S.c. and intrathecal injection of SCF markedly reduced the paw withdrawal threshold to mechanical stimuli and intrathecal SCF at 10 pg maximally induced mechanical allodynia in conscious mice. Intrathecal SCF also reduced the paw withdrawal latency to heat stimuli significantly but transiently. The c-kit receptor was co-expressed in 58.4% of calcitonin gene-related peptide (CGRP) -positive, but only 5.1% of isolectin B4-positive, DRG neurons. In the spinal cord, the c-kit receptor was detected in the superficial layer of the dorsal horn and co-localized there with CGRP in central terminals of DRG neurons. Selective elimination of unmyelinated C-fibers by neonatal capsaicin treatment resulted in marked reduction of the c-kit receptor and CGRP expression in the superficial layer of the spinal cord. Cell-size profiles showed that c-kit receptor expression was significantly up-regulated and down-regulated in medium-sized DRG neurons after neonatal capsaicin treatment and nerve injury, respectively. These results suggest that the c-kit receptor is mainly expressed in peptidergic small-sized DRG neurons and may be involved in pain regulation both peripherally and centrally.

Pseudomonas fluorescens HYK0210-SK09 offers species-specific biological control of winter algal blooms caused by freshwater diatom Stephanodiscus hantzschii.

AIMS: The study of an algicidal activity and mechanism of the isolated Pseudomonas fluorescens HYK0210-SK09 (SK09) against a winter bloomed harmful diatom, Stephanodiscus hantzschii. METHODS AND RESULTS: SK09 was isolated from the Paldang reservoir, Korea and used to biological control of S. hantzschii. The inoculation of SK09 at the final density of 5 x 10(6) cells ml(-1) caused degradation of >90% of S. hantzschii cells within 5 days. The algal cell lysis was achieved by a direct attack of the bacteria to the diatom cells, and the algicidal compound was located in the cytoplasm of the cell. As SK09 did not suppress Microcystis aeruginosa, Anabaena cylindrica, Coelastrum astroideum or Cyclotella meneghiniana, it appeared to attack S. hantzschii in a species-specific manner. Testing in an indoor mesocosms confirmed that SK09 effectively reduced S. hantzschii cells by 88% within 9 days. CONCLUSIONS: This bacterium is useful in regulating blooms of S. hantzschii. However, it should be studied in the future that their impact in shaping phytoplankton community and their activity in natural environments. SIGNIFICANCE AND IMPACT OF THE STUDY: The bacterium enabled us to develop a new strategy, to understand the interaction for anthropogenic control of harmful algal blooms in nature.

Transition of Spleen Volume Long After Pediatric Living Donor Liver Transplantation for Biliary Atresia.

PURPOSE: After undergoing the Kasai procedure for biliary atresia (BA), most patients develop severe splenomegaly that tends to be improved by liver transplantation. However, fluctuations in splenic volume long after transplantation remain to be elucidated. PATIENTS AND METHODS: Seventy-one consecutive patients who had undergone pediatric living donor liver transplantation (LDLT) for BA were followed up in our outpatient clinic for 5 years. They were classified into 3 groups according to their clinical outcomes: a good course group (GC, n = 41) who were maintained on only 1 or without an immunosuppressant, a liver dysfunction group (LD, n = 18) who were maintained on 2 or 3 types of immunosuppressants, and a vascular complication group (VC, n = 11). Splenic and hepatic volumes were calculated by computed tomography in 464 examinations and the values compared before and after the treatment, especially in the VC group. RESULTS: Splenic volume decreased exponentially in the GC group, with splenic volume to standard spleen volume ratio (SD) being 1.59 (0.33) 5 years after liver transplantation. Splenic volume to standard spleen volume ratios were greater in the VC and LD groups than in the GC group. Patients in the VC group with portal vein stenosis developed liver atrophy and splenomegaly, whereas those with hepatic vein stenosis developed hepatomegaly and splenomegaly. Interventional radiation therapy tended to improve the associated symptoms. CONCLUSIONS: Fluctuations in splenic volume long after pediatric LDLT for BA may reflect various clinical conditions. Evaluation of both splenic and hepatic volumes can facilitate understanding clinical conditions following pediatric LDLT.

Antibody Drug Treatment for Steroid-Resistant Rejection After Pediatric Living Donor Liver Transplantation: A Single-Center Experience.

BACKGROUND: Antibody drugs have been used to treat steroid-resistant rejection (SRR) after liver transplantation. Although anti-thymocyte globulin has been used for SRR after liver transplantation in place of muromonab-CD3 since 2011 in Japan, the effectiveness of anti-thymocyte globulin after pediatric living-donor liver transplantation (LDLT) has not yet been reported. The aim of this study was to evaluate the effectiveness of antibody drug treatment for SRR after pediatric LDLT in our single center. METHODS: Between May 2001 and December 2013, 220 pediatric LDLTs were performed. Initial immunosuppression after LDLT included tacrolimus and methylprednisolone therapy. Acute rejection was diagnosed by use of a liver biopsy and the administration of steroid pulse treatment, and SRR was defined as acute rejection refractory to the steroid pulse treatment. RESULTS: Acute rejection and SRR occurred in 74 (33.6%) and 16 patients (7.3%), respectively. The graft survival rates of non-SRR and SRR were 92.4% and 87.5%, respectively (P = .464). The median concentration of alanine aminotransferase before and after the administration of antibody drug was 193.5 mU/mL (range, 8-508) and 78 mU/mL (range, 9-655), respectively (P = .012). The median rejection activity index before and after the administration of antibody drugs was 5 (range, 2-9) and 1 (range, 0-9), respectively (P = .004). After antibody drug treatment, 12 patients had cytomegalovirus infections, 2 patients had Epstein-Barr virus infections, 3 patients had respiratory infections, and 1 patient had encephalitis. The cause of death in 1 patient with SRR was recurrence of infant fulminant hepatic failure. CONCLUSIONS: Antibody drug treatment for SRR after pediatric LDLT is safe and effective.

Endotoxin Metabolism Reflects Hepatic Functional Reserve in End-Stage Liver Disease.

BACKGROUND: The hepatic clearance of endotoxin (Et) may reflect hepatic functional reserve and ischemic injury to hepatocytes. Therefore, we examined the relationships between Et activity (EA) and the metrics Pediatric End-Stage Liver Disease (PELD)/Model of End-Stage Liver Disease (MELD) score and alanine transaminase (ALT) levels in the postoperative period. METHODS: We performed 8 living-donor liver transplantations (LDLTs) for biliary atresia at our center from April 2012 to December 2012. EA was measured by means of an Et activity assay (EAA) in samples collected from a vein 1 day before LDLT, from the portal vein during the intraoperative anhepatic phase, from an artery 1 hour after reperfusion, from an artery on postoperative day (POD) 1, and from an artery or vein at PODs 7 and 14. RESULTS: EAs generally remained at low levels. EA at the reperfusion period was significantly lowest. The correlation coefficient for the preoperative MELD/PELD score and the EAA was 0.837, and the corresponding P value was .009; thus, there was a significant relationship between the preoperative MELD/PELD score and the EAA. The correlation coefficients for ALT at POD 1 and EA during the anhepatic phase, at 1 hour after reperfusion, and at POD 1 were 0.64, 0.43, and 0.38, respectively, and the P values for these correlations were .08, .67, and .34. Thus, we observed that ALT and EA generally tended to be somewhat directly correlated, but no significant relationships between these 2 metrics were observed. CONCLUSIONS: Endotoxin metabolism reflects the hepatic functional reserve capacity of end-stage liver disease.

Elucidation of the mechanism of suppressed steroidogenesis during androgen deprivation therapy of prostate cancer patients using a mouse model.

Androgen deprivation therapy (ADT) is the standard medical approach to the management of prostate cancer. Patients switched from a GnRH antagonist to a GnRH agonist, did not experience a testosterone surge in spite of the occurrence of luteinizing hormone (LH) surge in our protocol of clinical study. To clarify this observation, male mice pre-treated with two different doses of the GnRH antagonist degarelix for 28 days were further administered the GnRH agonist leuprolide or chorionic gonadotropin, and testosterone production of the mice was studied. Serum LH and testosterone levels, the size of Leydig cells, and expression level of steroidogenesis-related genes in the testis were analyzed. Treatment of mice with a high dose of degarelix (0.1 µg/mouse; HDG), but not a low dose (0.05 µg/mouse; LDG), for 28 days reproduced declined steroidogenesis observed in prostate cancer patients during ADT switched from a GnRH antagonist to a GnRH agonist. The size of the Leydig cells in the HDG mice was not significantly different from that in naive mice. Although expression levels of StAR, P450scc, and 17ß HSD increased significantly in the LDH testis, those in the HDG testis did not change. Treatment of mice with a high dose of degarelix for 28 days reproduced the decline in steroidogenesis observed in prostate cancer patients during ADT. In this animal model, we demonstrated that initial ADT may inhibit the ability of Leydig cells to produce testosterone by suppressing the expression of genes involved in steroidogenesis, such as StAR, P450scc, and 17ßHSD.

Relationship Between Graft Liver Function and the Change of Graft Liver and Spleen Volumes After Technical Variant Liver Transplantation.

BACKGROUND: Although there have been a few reports describing the changes of graft liver and spleen volumes after liver transplantation (LT), little is known about the relationship between graft liver function and the changes of these volumes after technical variant liver transplantation (TVLT). We therefore performed a retrospective study to investigate the relationship between graft liver function and these volumes after TVLT. METHODS: We retrospectively investigated the cases of 140 TVLT procedures that were performed in our department between July 1987 and October 2012 and in which follow-up was conducted at our department. We calculated the graft liver volume to standard liver volume (GV/SLV) ratio, the spleen volume to standard spleen volume (SV/SSV) ratio, and the spleen volume to graft liver volume (S/L) ratio by CT volumetry. We clarified the relationship between graft liver function (according to the pathological findings) and the graft liver and spleen volumes at 2, 5, and 10 years after TVLT. RESULTS: In the normal liver function group, the GV/SLV, SV/SSV, and S/L ratios decreased until 6 months after TVLT and then converged at 10 years after TVLT to 0.95, 1.27, and 0.27, respectively. In the graft liver failure group, the GV/SLV, SV/SSV, and S/L ratios at 10 years after TVLT were 0.67, 5.01, and 1.55, respectively. A significant correlation was observed between the GV/SLV ratio and the presence of mild liver fibrosis at 2 and 5 years after TVLT (P = .03 and P = .04, respectively). CONCLUSIONS: Post-transplant CT-volumetry is a noninvasive and effective means of evaluating graft liver status.

An efficient expression system for a variant form of the cytotoxic protein alpha-sarcin in Escherichia coli.

An efficient Escherichia coli system for the production of a variant form of the cytotoxic protein alpha-sarcin (delta Ala 1) has been constructed. cDNA encoded alpha-sarcin lacking N-terminal alanine was ligated with the bla signal peptide sequence (the signal sequence of E. coli beta-lactamase localizes the protein in the periplasm) and was inserted into an inducible bacterial expression vector pKTN2-2. When the plasmid introduced into E. coli was expressed in the presence of IPTG, the recombinant product accumulated in the periplasmic space. The product was purified by Affi-Gel Blue followed by Bio-Rex 70 column chromatography. Recombinant alpha-sarcin (delta Ala 1) displayed the properties similar to those of authentic alpha-sarcin isolated from Aspergillus giganteus with respect to its molecular mass and enzymatic activity in ribosomal inactivation. The amount of alpha-sarcin variant produced in the system was estimated to be 1.2 mg/l of culture.

Involvement of Rho-kinase in inflammatory and neuropathic pain through phosphorylation of myristoylated alanine-rich C-kinase substrate (MARCKS).

Myristoylated alanine-rich C-kinase substrate (MARCKS) is a major in vivo substrate for protein kinase C in the brain and has been implicated in cellular processes associated with cytoskeletal restructuring such as synaptic trafficking and neurotransmitter release. A phosphorylation-site specific antibody against Ser159-phospho-MARCKS (pS159-Mar-Ab) revealed that MARCKS is phosphorylated at Ser159 by Rho-kinase and that its phosphorylation is inhibited by the Rho-kinase specific inhibitor H-1152. Since the function of MARCKS is regulated by phosphorylation at multiple sites, here we examined the involvement of Rho-kinase in relation to phosphorylation of MARCKS at Ser159 in inflammatory and neuropathic pain by H-1152. When intrathecally administered 10 min before s.c. injection of formalin, H-1152 at 10 and 100 ng attenuated the second-phase, but not the first-phase, pain-like behaviors in the formalin test. Neuropathic pain induced by selective L5 spinal nerve transection was also relieved by intrathecal injection of H-1152. Nitric oxide synthase activity visualized by NADPH diaphorase histochemistry increased in the superficial layer of the spinal cord 30 min after formalin injection and 7 days after nerve transection, which were blocked by H-1152. Phosphorylation of MARCKS at Ser159 was detected in the spinal cord by pS159-Mar-Ab and the level of phosphorylation increased in the superficial layer after nerve transection. In contrast, immunoreactivities of neuronal nitric oxide synthase and MARCKS did not change significantly in the spinal cord before and after nerve transection. Taken together, the present study demonstrates that Rho-kinase is involved in inflammatory pain and the maintenance of neuropathic pain through phosphorylation of MARCKS at Ser159.

Surgically induced detachment of the anterior hyaloid membrane from the posterior lens capsule.

Vitreous hemorrhage adhering to the posterior lens capsule prevents adequate visualization of the vitreous cavity and fundus during vitreous surgery and during the dissection of fibrovascular membranes. This type of hemorrhage is difficult to remove by aspiration or resection using a vitreous cutter. We have developed a new technique designed to detach surgically the anterior vitreous for the removal of hemorrhage in patients with proliferative diabetic retinopathy. In this hydrodissection technique, the anterior vitreous is detached from the posterior lens capsule by a forced injection of infusion fluid into the anterior chamber. This technique separates the vitreous hemorrhage adhering to the posterior lens capsule and allows its removal.

Attached posterior hyaloid membrane and the pathogenesis of honeycombed cystoid macular edema in patients with diabetes.

PURPOSE: To investigate the pathogenesis of honeycombed cystoid macular edema in patients with diabetes. METHODS: The relationship between the posterior hyaloid membrane and honeycombed cystoid macular edema was examined in 24 eyes of 20 patients with diabetes who underwent vitrectomy for cystoid macular edema. RESULTS: In 19 eyes of 15 patients with diabetes, the posterior hyaloid membrane was attached to the macula, and in five eyes of five patients, the posterior hyaloid membrane was separated from the macula. In 14 (74%) of the 19 eyes with an attached posterior hyaloid membrane, honeycombed cystoid macular edema was detected. Conversely, honeycombed cystoid edema was not detected in any of the five eyes with posterior hyaloid detachment (P = .0059). CONCLUSIONS: These observations demonstrate a strong correlation between an attached posterior hyaloid membrane and the presence of honeycombed cystoid macular edema. We suggest that retinal traction by the posterior hyaloid membrane is involved in the pathogenesis of honeycombed cystoid changes in diabetic patients.

Examination of patients with cystoid macular edema using a scanning laser ophthalmoscope with infrared light.

PURPOSE: To report a noninvasive method for evaluating eyes with cystoid macular edema. METHODS: We obtained infrared images of cystoid macular edema in eight eyes of eight patients using a scanning laser ophthalmoscope with the dark-field mode of a 780-nm diode laser. Differences between infrared images and fluorescein angiograms in the imaging of cystoid changes were examined. RESULTS: With the scanning laser ophthalmoscope, we observed cystoid macular changes as images that resembled three-dimensional pictures in the dark-field mode with infrared light. Cystoid changes observed by this method generally agreed with changes observed by fluorescein angiography. CONCLUSIONS: Scanning laser ophthalmoscopy with infrared light in a dark-field mode is noninvasive, and the results in eyes with cystoid macular edema generally agreed with results obtained by fluorescein angiography. This method is useful for examining eyes with cystoid macular edema.

Vitrectomy for cystoid macular oedema with attached posterior hyaloid membrane in patients with diabetes.

AIM: To report the success of vitrectomy in eliminating cystoid macular oedema and improving vision in three eyes of two patients with diabetic cystoid macular oedema. In all of the eyes there was no ophthalmoscopic evidence of traction from a posterior hyaloid membrane or from proliferative tissue. METHODS: Pars plana vitrectomy was performed on three eyes of two patients with diabetic cystoid macular oedema who did not show traction upon examination with a slit lamp biomicroscope and a scanning laser ophthalmoscope. RESULTS: Cystoid changes disappeared 1, 3, and 5 days, postoperatively, and diffuse macular oedema resolved within 2 weeks. The visual acuity was improved and maintained. CONCLUSION: Vitrectomy can be effective in some patients with diabetic cystoid macular oedema even in patients who lack evidence of traction by ophthalmoscopy.

Streptokinase gene variable region classification in streptococci: lack of correlation with post-streptococcal glomerulonephritis.

To investigate a possible causal role of streptokinase (SKase) in acute post-streptococcal glomerulonephritis (APSGN), the major variable region of SKase genes of Streptococcus pyogenes strains isolated from patients with and without APSGN were analyzed using the polymerase chain reaction, restriction enzyme analysis and the direct sequencing of SKase genes. In the APSGN-associated strains, six of nine revealed mutant classes corresponding to the nephritogenic classes I and II proposed by Johnston et al. [1992], the remaining three belonged to non-nephritogenic classes. In twenty strains not associated with APSGN, seventeen belonged to classes I and II, while three were from other classes. The major variable region of the SKase gene shows no apparent relation with induction of APSGN in humans, suggesting that unique classes of streptococcal SKase do not play a role in the pathogenesis of APSGN.

Preparation of drug-diluent hybrid powders by dry processing.

New hybrid powders have been produced by the dry processing of six drugs (oxyphenbutazone, prednisolone, theophylline, indomethacin, phenacetin and aspirin), with potato starch used as a core material, by means of an electric mortar and a powder surface reforming system designed to produce hybrid powders. The hybrid powders obtained immediately after production differed in their structure from interactive mixtures. With the hybrid powders the drug was spread on the surface of the core particle by friction and collision that occurred in the dry process, but with interactive mixtures the drug simply adhered as intact particles to the surface of diluent particles. Scanning electron microscopy and powder X-ray diffractometry indicated that the mechanochemical phenomenon was essential for the production of the hybrid powders. With time, a shape change in the adhering drug was observed as a relaxation process took place, with recrystallization resulting from the release of accumulated energy. The change with time might depend upon the method of producing powders and the physical properties of the drug used, e.g. the smooth layer of indomethacin produced by the powder surface reforming system reverted to fine particles tightly adhering to the starch surface, though no change was observed with prednisolone.

[Meningioma associated with acute subdural hematoma: a case report].

We present a case of meningioma associated with acute subdural hematoma. This 67-year-old male had a sudden onset of severe headache when he was on the train. He had a CT scan which revealed an acute subdural hematoma at the left parietal convex. Cerebral angiography disclosed a small focus (3 x 4 cm) of vascular stain under the left parietal bone supplied by the left middle meningeal artery. He was diagnosed as having a meningioma with surrounding acute subdural hematoma. The removal of this tumor was carried out without delay. It was fragile and the bleeding point was not detected. Pathological diagnosis was meningothelial meningioma. The literature showed meningioma associated with acute subdural hematoma is rare, but when it is discovered incidentally, surgical resection might be indicated.

[A case of chronic neutrophilic leukemia associated with polycythemia vera].

A 74-year-old male patient was seen on December 15, 1980 because of right shoulder pain and leukocytosis. The spleen and the liver were enlarged, and palpable, 3 and 4 fingerbreadths below the costal margin respectively. The red blood cell count (RBC) was 899 x 10(4)/microliters, hemoglobin (Hb) 20.6 g/dl, reticlocyte (Ret) 7/1000, platelets (Plt) 34.2 x 10(4)/microliters, the white blood cell count (WBC) 26,800/microliters with 86% neutrophils. A bone marrow aspiration showed the nucleated cell count (NCC) to be 16.5 x 10(4)/microliters, and the myeloid/erythroid ratio (M/E) 2.0. The patient was treated with venesection and later with carboquone. Since September 1990 he has not been treated with any agents because of erythrocytopenia. In July 25, 1991, the spleen was enlarged, and palpated 8 fingerbreadths below the costal margin. The RBC was 456 x 10(4)/microliters, Hb 12.5 g/dl, Ret 9/1000, Plt24.2 x 10(4)/microliters, the WBC41,500/microliters with 90% neutrophils. Bone marrow aspiration showed the NCC to be 16.5 x 10(4)/microliters, M/E14.3, with no atypical cells. Chromosome studies of marrow cells revealed no Ph1 chromosome. Many hematologic malignancies have been occasionally associated with polycythemia vera (PV), but chronic neutrophilic leukemia associated with PV is a very rare condition.