Monoclonal antibodies for radioimmunoscintigraphy of breast cancer

Breast cancer is the leading cause of cancer deaths among females, and it is estimated that each year, one in ten American women will be newly diagnosed as having the disease. It is therefore not surprising, that a great deal of effort has been made to better understand the biology of breast cancer, and that investigators keep up the search for new tools to better characterize, diagnose and treat these tumours. In this regard, the introduction of the hybridoma technique in 1975 by Kohler and Milstein has lead to an extensive work in the characterization of monoclonal and polyclonal antibodies against breast cancers. A large number of antibodies has been raised to different epitopes present in normal and neoplastic breast tissue; but unfortunately we have yet to find a highly sensitive and specific monoclonal antibody for breast cancer that can successfully be used for scintigraphic detection of nodal metastases and for radioimmunotherapy treatment of this disease.As possible radioimmunodiagnostics, antibodies are known which react with the following antigens:

• 1.(1) cytoskeletal proteins
• 2.(2) breast cell products
• 3.(3) steroid receptors
• 4.(4) putative tumor-associated antigens
• 5.(5) oncogene products
• 6.(6) pregnancy-related products
• 7.(7) basement membrane antigens
• 8.(8) degradative enzymes
• 9.(9) cell receptors for extracellular matrix molecules
• 10.(10) multidrug resistance gene product (p-glycoprotein)
• 11.(11) proliferative markers.

     

Isohematoporphyrin-diesters and diethers

• 1.1. A series of diesters of isohematoporphyrin (isoHp), from dimethyl to dioctyl were prepared according to Rimington et al. (1989b). Their optical absorption, fluorescence spectra and high performance liquid chromatography (HPLC) retention times were recorded.
• 2.2. A plot of HPLC retention time against number of C atoms in the alcohol used for esterification was approximately linear at first then rising steeply from diamyl to diocyi ester, whether a gradient elution was used or only methanol: water, 95/5, at pH 7.5.
• 3.3. Preparation of the diethers of isoHp was much more difficult than that of the corresponding derivatives of hematoporphyrin (Hp). Several different methods were investigated, varying both times and temperatures.
• 4.4. These methods included reaction of isoHp or its demethyl ester with
  • 4.1.(i) a bromoalkane in presence of anhydrous K₂CO₃;
  • 4.2.(ii) reaction with bromoalkane and Ag₂O;
  • 4.3.(iii) reaction of brominated-isoHp, prepared by using thionylbromide, with the selected alcohol, or corresponding sodium alcoholate;
  • 4.4.(iv) heating of isoHp alone with an alcohol containing 20% (w/v) H₂SCO₄ (temp. range from 45° to 118°C),
  • 4.5.(v) refluxing as in (iv) at the b.p. of the alcohol; and
  • 4.6.(vi) carrying out this reaction in refluxing ethyleneglycoldimethyl ether (b.p. 85°C) or diethyleneglycoldimethyl ether (b.p. 155°C).
• 5.5. Some diether formation was observable by all these methods but yields were small, a considerable quantity of unreacted isoHp and other products remaining.
• 6.6. Examined by HPLC, the diethers consistently afforded a forked peak which on thin layer chromatography was only resolved into two very closely associated bands by a solvent mixture carefully selected for development.
• 7.7. On elution these materials had virtually identical optical absorption and fluoresence spectra.
• 8.8. The nature of the association is discussed, atropisomers (Gottwald and Ullman, 1969) and possible stacked monomer: dimers (Abraham et al., 1963) being considered as possibilities.

     

Biomines-stimulated phosphorylation and (Na+, K+-ATPase in the gills of the chinese crab,Eriocheir sinensis

• 1.1. Subcellular distribution of (NA⁺, K⁺-ATPase and ouabain-insensitive ATPase (Mg²⁺-ATPase) are compared in branchial tissues of the euryhaline crab, Eriocheir sinensis, acclimated to fresh water.
• 2.2. Both the anterior and posterior gills contain cAMP-dependent protein kinase and endogenous protein substrate for phosphorylation.
• 3.3. Phosphorylation occurs in both “particulate” and “soluble” subcellular fractions but its stimulation by cAMP is restricted to the “soluble” fraction.
• 4.4. serotonin (5-HT) and dopamine receptors are present only in the “light particulate” fraction isolated from the posterior gills.

• 1.(a) Serotonin and dopamine have no effect on the phosphorylation observed in a subcellular fraction alone.
• 2.(b) Activation of the phosphorylation by serotonin and dopamine is found when the soluble fraction (source of cAMP-dependent protein kinase) is added to the fraction P₃ from the posterior gills.
• 3.(c) No activation occurs with the fractions P₃ as well as P₁ or P₂ (not shown) from anterior gills of fresh water crab.
• 4.(d) Cyproheptadine, a serotonin receptor antagonist, inhibits the 5-HT dependent increase in phosphorylation.
• 5.(e) The dopamine receptor antagonist, chlorpromazine, inhibits dopamine-stimulated phosphorylation.
• 6.5. Ouabain mimics the effect of cyproheptadine on the serotonin-stimulated phosphorylation found in the posterior gills.

     

Functional groups in the social behavior of a cichlid fish,the Oscar,Astronotus ocellatus

Dummy conspecifics were presented to isolated adults of the cichlid fish Astronotus ocellatus to investigate the functional organization of cichlid social behavior. Body size and 15 dummy-elicited activities were recorded during 15 min sessions and analyzed by principal components analysis (PCA) to reveal their temporal organization. Five principal components explained almost 80% of the variation in dummy-elicited behavior, and these five factors define functional groups for

• 1.(a) investigation,
• 2.(b) attack,
• 3.(c) nesting,
• 4.(d) boldness,
• 5.(e)distress.

Nest-oriented and attack modal action patterns are not mutually inhibitory during this time frame, and biting does not appear to function exclusively during an attack on a conspecific. Comparison with previous studies of New and Old World cichlids suggests evolutionary conservation of the functional organization of social behavior.     

Ecdysone-controlled meiotic reinitiation in oocytes of Locusta migratoria involves a decrease in cAMP levels

Evidence for the involvement of cAMP in the triggering of meiotic reinitiation by ecdysone in vitellogenic oocytes of Locusta migratoria is presented:

• 1.(1) the intracellular concentration of cAMP decreases significantly (by 40%) in the oocytes at the time when meiotic reinitiation is induced;
• 2.(2) drugs which increase the concentration of cAMP antagonize the stimulatory action of ecdysone;
• 3.(3) ecdysone treatment of excised oocytes is followed by a decrease in intra-cellular cAMP;
• 4.(4) ecdysone reduces the adenylate cyclase activity when added to plasma membrane preparations in vitro.

     

Tritiated water measurements of water flux in nestling and adult zebra finches (Poephila guttata)

• 1.1. Total body water volume (TBWV) and daily water turnover rates (WTR) were measured in nestling and adult zebra finches using tritiated water (TOH).
• 2.2. TBWV and daily WTR increased with age up to 13 days post hatching.
• 3.3. TBWV and WTR approached adult levels after 13 days of age. WTR varied among ages and nest mates. All nestlings turned over at least 50% of their body water pool per day.
• 4.4. The WTR data for adult birds are consistent with natural history data suggesting zebra finches are dependent on water for breeding and survival.

     

The changes in body temperature,oxygen consumption,CO2 production and muscle protein turnover rate by selection for body size in Japanese quail,Coturnix coturnix japonica

• 1.1. Body temperature, oxygen consumption, CO₂ production and muscle protein degradation rate were measured in the three quail lines selected for body size, a random bred line (RR) and two lines selected for large (LL) or small (SS) body size.
• 2.2. The body temperature at 15 weeks of age was highest for small body size line and lowest for large body size line.
• 3.3. The body temperature, oxygen consumption and CO₂ production of females were significantly higher than that of males.
• 4.4. The fractional degradation rate of muscle protein of SS, RR and LL lines were measured as 2.4, 1.6 and 1.2% per day in male, and 2.6, 1.7 and 1.4% per day in female.

     

Sensitive Determination of Proteolytic Proteoforms in Limited Microscale Proteome Samples

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Highlights

• •Single-pot workflow for manual or automated enrichment of N-terminal peptides.
• •Sensitive enrichment of protein N termini from 10,000 cells or 2 μg crude proteome.
• •Data independent acquisition improves precision of peptide level quantification.
• •First degradomic analyses of sorted immune cells, single seedlings, and mitochondria from patient cells.

     

N-glycopeptide Signatures of IgA2 in Serum from Patients with Hepatitis B Virus-related Liver Diseases

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Highlights

• •¹⁸O/¹⁶O labeling N-glycopeptide quantification for 40-kDa band in HCC and LC.
• •MRM verification of aberrant N-glycopeptides in healthy-HBV-LC-HCC cascade.
• •TPLTAN²⁰⁵ITK (H5N5S1F1) and (H5N4S2F1) of IgA₂ change in liver diseases.
• •Variation in two N-glycopeptides abundance not caused by protein concentration.

     

Glycoproteogenomics: A Frequent Gene Polymorphism Affects the Glycosylation Pattern of the Human Serum Fetuin/α-2-HS-Glycoprotein

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Highlights

• •Frequent genetic polymorphism affects the glycosylation pattern of fetuin.
• •Personalized in-depth proteoform profiling of fetuin purified from 20 donors.
• •Classification of serum donors into three different genotypes.
• •Septic patients show increased level of fucosylation at N-glycolation site N176.

     

Breast,hips, and buttocks revisited: Honest fatness for honest fitness

This paper comments on: Low, B. S., Alexander, R. D., and Noonan, K.M. Human hips, breast, and buttocks: Is fat deceptive? Ethology and Sociobiology 8: 249-247, 1987. In it I argue that:

• 1.1. Sexual selection has probably not been the most important selection pressure on
• 2.female human body shape.
• 3.2. Male humans in different cultures find different aspects of the female body attractive
• 4.and therefore are unlikely to have exerted consistent directional sexual selection on
• 5.the female body.
• 6.3. Breast size is not correlated with lactation success.
• 7.4. Visible hip width is not correlated with parturition success.
• 8.5. Women would lower their fitness if they tried to deceive men about their internal
• 9.pelvic dimensions.
• 10.6. There are many alternative hypothesis to explain the existence of fat onwomen's
• 11.breast, hips, and buttocks.

     

The tolerance of lepidopterous larvae to an insect selective neurotoxin

The interaction of the fast-neurotoxic and insect selective polypeptide derived from scorpion venom (AaIT) with lepidopterous larvae tissues was studied through assays of toxicity, chromatography, binding and light microscopical autoradiography. The native and/or radioiodinated toxin was shown to:

• 1.(1) Induce a delayed, slow, progressive paralysis (within 24–48 h) of Spodoptera larvae by relatively high doses (paralytic unit = 2.4 μg/100 mg) corresponding to about only 10% of the total toxicity of the crude venom. Larvae of six species representing five families of Lepidoptera responded similarly to the toxin.
• 2.(2) Resist an in vitro incubation in the insect's hemolymph.
• 3.(3) Lose 80% of its toxicity in the insect's body within 24 h, accompanied by a progressive process of degradation and elimination by the excretory system.
• 4.(4) Specifically bind to a single class of non-interacting binding sites of high affinity and low capacity (0.2 pmol/mg protein, similar to tritiated saxitoxin) in an in vitro, homogenate derived, neuronal preparation.
• 5.(5) Specifically bind with high affinity to desheathed but otherwise intact nerves.
• 6.(6) Be devoid of accessibility to peripheral-terminal branches of Spodoptera motor nerves in situ—strongly contrasting those of the toxin susceptible Periplaneta nerves.

It may be thus concluded that the tolerance of the lepidopterous larvae to AaIT can be substantially attributed to pharmacokinetic aspects of toxin accessibility barriers and degradation processes.     

Electric organ discharge frequency modulation evoked by water vibration in Gymnotus carapo

• 1.1. The electric organ discharge (EOD) frequency modulations evoked by brief water vibration were analysed in the pulse-type fish Gymnotus carapo.
• 2.2. The response consisted of a transient increase of the EOD frequency at short latency (30 msec). Response profiles were characteristic of the specimen and relatively independent on stimulus intensity.
• 3.3. Conversely, they were dependent on stimulation sequence, showing a rapid decrement along successive stimuli and high temporal discrimination.
• 4.4. The brief latencies indicate a relatively simple neural circuit.
• 5.5. The response may be an electrolocation enhancement strategy for the detection of moving objects based on “sampling” the periphery at a higher frequency.

     

Secretory oedema in diabetic submandibular glands during parasympathetic nerve stimulation: Relationship to microvascular abnormalities in streptozotocin-treated rats

• 1.1. Submandibular secretion during parasympathetic stimulation (5 Hz) was examined in streptozotocin-diabetic and age-matched control rats.
• 2.2. At 3 weeks, but not 3 and 6 months, flow rate was initially greater than in controls, but it declined rapidly after 30 min.
• 3.3. The reduction in flow rate was associated with oedema of the gLond.
• 4.4. At 3 months, graded stimulation revealed a tendency to oedema at frequencies of 10 Hz and above.
• 5.5. Morphologically, submandibular capillary density was increased in diabetic rats.
• 6.6. Thus, in diabetes the submandibular gland appears less able to withstand continuous parasympathetic stimulation, due in part to an increase in tissue capillary area.

     

Synaptic transmission at high pressure: Effects of [Ca2+]o

• 1.1. The effects of pressure on synaptic currents were examined in crayfish abdominal muscles.
• 2.2. Helium pressure (10.1 MPa) considerably decreased extracellulariy-recorded excitatory junctional potentials associated with increased short-term facilitation.
• 3.3. These effects could be mimicked by a reduction of [Ca²⁺]_o, and partially compensated by an increase in [Ca²⁺]_o.
• 4.4. Pressure also reduced the amplitude of the extracellular nerve terminal potentials (ENTP) by up to 25%, and significantly increased synaptic delay in a [Ca²⁺]_o-dependent manner.
• 5.5. The interaction between compression and various [Ca²⁺]_o were analysed in terms of an existing model of transmitter release. The results were consistent with the hypothesis that high pressure decreases the maximal Ca²⁺ influx into nerve terminals.
• 6.6. The decreased ENTP and increased synaptic delay suggest that additional processes may be involved in pressure effects on synaptic transmission.

     

Identification of neurons involved in the earthworm Amynthas hawayanus reflex activity

• 1.1. The medial (MGF), lateral (LGF) and motor (RMS-2) giant neurons were confirmed as neural components in the earthworm Amynthas hawayanus polysynaptic reflex circuit by simultaneous potential recording and dye injection.
• 2.2. The reflex was initiated from the mechanoreceptors when evoked by mechanical stimulation but electrical stimulation also evoked an antidromic response in the motoneuron.
• 3.3. The primary reflex response propagates decrementally along both giant axons but directly evoked action potentials conduct in an all-or-none fashion.
• 4.4. The secondary reflex response continues to propagate after the primary response disappears.
• 5.5. A rhythmically discharging neuron of uncertain function was also identified.

     

Proteome-wide Tyrosine Phosphorylation Analysis Reveals Dysregulated Signaling Pathways in Ovarian Tumors

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Highlights

• •Multiplexed PTM assays on HuProt array were developed using ovarian tumor lysates.
• •Proteome-wide Tyr phosphorylation with 102 ovarian tumors were performed and analyzed.
• •19 kinases were predicted to have elevated activities in ovarian tumor.
• •Elevated activities of PTK2 and PTK2B were confirmed in ovarian cancer cell lines.

     

Changes in body composition of germ-free and conventional chickens during starvation

• 1.1. Changes in body composition during starvation were compared between germ-free (GF) and conventionalized (CVL) chicks in experiment 1. At 8 days of age, the GF birds were divided into two groups, i.e. GF and CVL groups. The CVL birds were inoculated with faeces from conventionally reared birds. Until 14 days of age, both birds were fed a diet ad lib, and thereafter starved for 6 days.
• 2.2. Nitrogen loss during starvation was significantly lower in CVL birds, though the reverse was true for water loss. Fasting heat production was comparable between two environments.
• 3.3. Influence of the gut microflora on body weight and nitrogen losses during starvation was investigated in birds prefed diets high or low in dietary protein in experiment 2.
• 4.4. No significant effect of the gut microflora was observed in body weight and nitrogen losses. Body weight was severely reduced in birds prefed the high protein diet and nitrogen loss was lower in birds prefed the low protein diet.

     

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Characterization of Endogenous Human FcγRIII by Mass Spectrometry Reveals Site,Allele and Sequence Specific Glycosylation*

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Highlights

• •Glycosylation of endogenous FcγRIII from neutrophils and matched plasma from more than 40 donors characterized at two sites involved in IgG binding.
• •Glycosylation of soluble FcγRIII glycosylation at N45 can be used to assign FcγRIIIb alleles.
• •FcγRIIIb allele specific differences in glycosylation at N162 may influence differential activity observed for primary cells.